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唐 智, 符 立, 张 焱, 周 博, 冯 天, 杨 文, 梁 鸽, 严 茜, 郑 璨, 别 明, 王 保. [Viable Bacteria Assay of Helicobacter pylori by RT-qPCR Measurement of cgt Gene Expression Levels: Establishment and Application of a New Method]. SICHUAN DA XUE XUE BAO. YI XUE BAN = JOURNAL OF SICHUAN UNIVERSITY. MEDICAL SCIENCE EDITION 2024; 55:1316-1321. [PMID: 39507989 PMCID: PMC11536244 DOI: 10.12182/20240960402] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 07/22/2024] [Indexed: 11/08/2024]
Abstract
Objective To establish a viable bacteria assay for Helicobacter pylori (H. pylori) by assessing the cgt gene expression, and to develop accordingly a rapid and novel testing method for clinical precision treatment. Methods Viable bacteria count was determined in bacterial cultures. The transcriptional expression level of cgt (hp0421), the conserved gene that encodes cholesterol-α-glucosyltransferase (CGT) in H. pylori, was measured by RT-PCR. The correlation between the number of colonies and cgt gene transcription expression was analyzed and the regression model was constructed. The linear range, sensitivity, and specificity of the new method were examined accordingly. The bactericidal action of clarithromycin was assessed using this method to verify the performance of the method in determining clinical bacterial drug resistance. Results The Ct values of cgt for H. pylori colony counts of 102, 104, 106, and 108 CFU/mL were 29.67±0.14, 23.37±0.36, 17.65±0.37, and 11.38±0.39, respectively. In the range of 101-108 CFU/mL, the regression equation for cgt gene expression and viable bacterial counts determined by RT-qPCR was y=-0.3501x+12.49, with the correlation coefficient being R 2=0.9992 and the sensitivity being 101 CFU/mL, showing no cross-reaction with 13 other bacteria. The lg values of live H. pylori bacteria treated with clarithromycin at 0, 5, 10, 20, and 40 μg/mL for 12 h were 2.57±0.02, 2.45±0.01, 2.19±0.02, 1.91±0.07, and 1.33±0.05, respectively. The corresponding cgt gene Ct values were 27.76±0.09, 28.37±0.24, 29.51±0.14, 30.11±0.12, and 31.66±0.11. By applying the cgt gene expression in the equation, the estimated counts of viable bacteria were found to be 2.73±0.03, 2.52±0.08, 2.11±0.05, 1.89±0.02, and 1.33±0.04, showing no significant difference in statistical analysis (P>0.05). Conclusion The method for assessing viable bacteria account by evaluating cgt gene expression in H. pylori was successfully established, significantly reducing the time required to determine viable bacteria count and providing a new method for clinical viable bacteria testing.
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Affiliation(s)
- 智慧 唐
- 四川大学华西基础医学与法医学院 （成都 610041）West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, Chengdu 610041, China
| | - 立发 符
- 四川大学华西基础医学与法医学院 （成都 610041）West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, Chengdu 610041, China
| | - 焱荣 张
- 四川大学华西基础医学与法医学院 （成都 610041）West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, Chengdu 610041, China
| | - 博彦 周
- 四川大学华西基础医学与法医学院 （成都 610041）West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, Chengdu 610041, China
| | - 天勤 冯
- 四川大学华西基础医学与法医学院 （成都 610041）West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, Chengdu 610041, China
| | - 文娟 杨
- 四川大学华西基础医学与法医学院 （成都 610041）West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, Chengdu 610041, China
| | - 鸽 梁
- 四川大学华西基础医学与法医学院 （成都 610041）West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, Chengdu 610041, China
| | - 茜雅 严
- 四川大学华西基础医学与法医学院 （成都 610041）West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, Chengdu 610041, China
| | - 璨璘 郑
- 四川大学华西基础医学与法医学院 （成都 610041）West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, Chengdu 610041, China
| | - 明江 别
- 四川大学华西基础医学与法医学院 （成都 610041）West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, Chengdu 610041, China
- 四川大学华西临床医学院 （成都 610041）West China School of Medicine, Sichuan University, Chengdu 610041, China
| | - 保宁 王
- 四川大学华西基础医学与法医学院 （成都 610041）West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, Chengdu 610041, China
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Wang Y, Liu L, Liu X, Wu K, Zhu X, Ma L, Su J. An Ultrasensitive PCR-Based CRISPR-Cas13a Method for the Detection of Helicobacter pylori. J Pers Med 2022; 12:jpm12122082. [PMID: 36556302 PMCID: PMC9784247 DOI: 10.3390/jpm12122082] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/21/2022] [Revised: 11/17/2022] [Accepted: 12/09/2022] [Indexed: 12/23/2022] Open
Abstract
The rapid and simple detection of Helicobacter pylori (H. pylori) is essential for its clinical eradication. Although various methods for detecting H. pylori have been well established, such as endoscopy in combination with histology or culture, rapid urease test (RUT) and molecular tests using clinical specimens, it is of great importance to develop an ultrasensitive and accurate nucleic acid detection platform and apply it to identify H. pylori. To meet these demands, a novel method based on PCR and CRISPR-Cas13a, called PCR-Cas13a, was developed and validated using the DNA of 84 clinical strains and 71 clinical specimens. PCR primers for the pre-amplification of conservative sequence and CRISPR RNA (crRNA) for the detection of specific sequence were designed according to the principle. The designed primers and crRNA were specific to H. pylori, and the assay showed a high degree of specificity compared with other common pathogens. Our detection system can screen H. pylori with a limit of 2.2 copies/μL within 30 mins after PCR amplification. Using a coincidence analysis with traditional methods, our method exhibited 100% accuracy for the detection of H. pylori. Furthermore, its diagnostic performance was compared, in parallel with a q-PCR. The PCR-Cas13a demonstrates 98% sensitivity and 100% specificity. Moreover, our approach had a lower limit of detection (LOD) than q-PCR. Herein, we present a diagnostic system for the highly sensitive screening of H. pylori and distinguish it from other pathogens. All the results demonstrated that this PCR-based CRISPR assay has wide application prospects for the detection of H. pylori and other slow-growth pathogens.
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Affiliation(s)
- Yaxuan Wang
- Department of Clinical Laboratory, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China
| | - Liyang Liu
- Department of Gastroenterology, Jingdong Medical Area, General Hospital of Chinese PLA, Beijing 101149, China
| | - Xiaochuan Liu
- Department of Gastroenterology, Emergency General Hospital, Beijing 100028, China
| | - Kai Wu
- Department of Gastroenterology, The Eighth Medical Center of PLA General Hospital, Beijing 100091, China
| | - Xiaoyan Zhu
- Department of Clinical Laboratory, The First Affiliated Hospital of Kunming Medical University, Kunming 650032, China
| | - Liyan Ma
- Department of Clinical Laboratory, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China
| | - Jianrong Su
- Department of Clinical Laboratory, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China
- Correspondence:
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Prevalence of Helicobacter pylori Virulence Genes and Their Association with Chronic Gastritis in Beijing, China. Curr Microbiol 2022; 80:33. [PMID: 36482124 DOI: 10.1007/s00284-022-03135-6] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/23/2022] [Accepted: 11/28/2022] [Indexed: 12/13/2022]
Abstract
Helicobacter pylori is closely related to chronic gastritis. The aim of the study was to investigate the correlation between H. pylori virulence genes and chronic gastritis in order to determine the pathogenic role of H. pylori virulence genes in chronic gastritis. Gastric mucosal tissues were obtained from 142 patients with chronic gastritis at three Beijing hospitals. The presence of virulence genes was determined by polymerase chain reaction (PCR) from H. pylori DNA. Multilocus sequence typing (MLST) and a phylogenetic tree were performed to characterize the overall genetic diversity. 91 new sequence types were identified by MLST in this study, and all strains showed high genetic diversity. The H. pylori isolates were divided into three types: hspEAsia strains (61 strains), hpEurope strains (15 strains), and mixed strains (16 strains). Some virulence genes were found to be significantly different between strains. The highest positive rates were found for dupA in chronic atrophic gastritis (AG), iceA1 in chronic non-atrophic gastritis with erosions, and iceA2 in chronic non-atrophic gastritis. The presence of dupA was found to be inversely related to the risk of AG. The H. pylori strains display high genetic diversity. Some virulence genes were found to be significantly different between diseases. The detection of various virulence genes is critical for screening high-risk populations for precancerous lesions and for the early prevention and control of gastric cancer.
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Ansari S, Yamaoka Y. Helicobacter pylori Infection, Its Laboratory Diagnosis, and Antimicrobial Resistance: a Perspective of Clinical Relevance. Clin Microbiol Rev 2022; 35:e0025821. [PMID: 35404105 PMCID: PMC9491184 DOI: 10.1128/cmr.00258-21] [Citation(s) in RCA: 33] [Impact Index Per Article: 11.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/07/2023] Open
Abstract
Despite the recent decrease in overall prevalence of Helicobacter pylori infection, morbidity and mortality rates associated with gastric cancer remain high. The antimicrobial resistance developments and treatment failure are fueling the global burden of H. pylori-associated gastric complications. Accurate diagnosis remains the opening move for treatment and eradication of infections caused by microorganisms. Although several reports have been published on diagnostic approaches for H. pylori infection, most lack the data regarding diagnosis from a clinical perspective. Therefore, we provide an intensive, comprehensive, and updated description of the currently available diagnostic methods that can help clinicians, infection diagnosis professionals, and H. pylori researchers working on infection epidemiology to broaden their understanding and to select appropriate diagnostic methods. We also emphasize appropriate diagnostic approaches based on clinical settings (either clinical diagnosis or mass screening), patient factors (either age or other predisposing factors), and clinical factors (either upper gastrointestinal bleeding or partial gastrectomy) and appropriate methods to be considered for evaluating eradication efficacy. Furthermore, to cope with the increasing trend of antimicrobial resistance, a better understanding of its emergence and current diagnostic approaches for resistance detection remain inevitable.
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Affiliation(s)
- Shamshul Ansari
- Department of Environmental and Preventive Medicine, Oita University Faculty of Medicine, Yufu City, Oita, Japan
| | - Yoshio Yamaoka
- Department of Environmental and Preventive Medicine, Oita University Faculty of Medicine, Yufu City, Oita, Japan
- Department of Medicine, Gastroenterology and Hepatology Section, Baylor College of Medicine, Houston, Texas, USA
- Institute of Tropical Disease, Universitas Airlangga, Surabaya, Indonesia
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Gastric Cancer Due to Chronic H. pylori Infection: What We Know and Where We Are Going. Diseases 2022; 10:diseases10030057. [PMID: 36135213 PMCID: PMC9498082 DOI: 10.3390/diseases10030057] [Citation(s) in RCA: 7] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/25/2022] [Revised: 08/21/2022] [Accepted: 08/24/2022] [Indexed: 11/28/2022] Open
Abstract
Helicobacter pylori is an established cause of many gastrointestinal pathologies including peptic ulcer disease, gastritis, and gastric cancer. It is an entity that affects the global population, and its true nature has only been known since the 1980s. Although there is much known about H. pylori including its pathophysiology, detection, and eradication, resistance to current therapy models is common. This is problematic because untreated or inadequately treated H. pylori increases morbidity and mortality related to gastric cancer and peptic ulcer disease among others. In order to improve the treatment and reduce resistance, there is significant ongoing research identifying new detection and eradication methods for H. pylori. This review aims to highlight what has already been established regarding H. pylori’s epidemiology, pathophysiology, detection, and treatment as well as the most current and novel research involving detection and treatment of H. pylori.
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Hsieh MS, Kuo FC, Wu MC, Wang JW, Liu CJ, Chu NS, Tsai PY, Hsu PI, Wu IC, Wu JY, Wu DC, Yu FJ, Kuo CH. Tailored susceptibility-guided therapy via gastric juice PCR for the first-line H. pylori eradication, a randomized controlled trial. J Formos Med Assoc 2021; 121:1450-1457. [PMID: 34836662 DOI: 10.1016/j.jfma.2021.10.011] [Citation(s) in RCA: 9] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/07/2021] [Revised: 07/22/2021] [Accepted: 10/18/2021] [Indexed: 12/18/2022] Open
Abstract
BACKGROUND/PURPOSE Clarithromycin-based standard triple therapy is still commonly adopted by 81.4% of physicians in real-world practice but yields low eradication rates. Therefore, we conducted this study to compare the efficacy of gastric juice-guided therapy for first-line eradication with the standard triple therapy, in order to provide an alternative to real-world practice. METHODS A total of 182 treatment-naïve Hp-infected patients were included and randomly allocated to either susceptibility-guided therapy (SGT) with gastric juice PCR or Clarithromycin-based standard triple therapy (STT) for 7 days. RESULTS The intention-to-treat eradication rates were 89% (81/91) in SGT and 75.8% in STT (p < 0.031). The per-protocol eradication rates were 91.0% (81/89) in SGT and 79.3% (69/87) in STT (p < 0.034). Among the subgroups of different antibiotic resistance, patients with SGT demonstrated superior eradication rates (91.7% vs 45.5%, p < 0.027) in the subgroup of both clarithromycin resistance and levofloxacin resistance. CONCLUSION This prospective randomized controlled trial demonstrated the reliable efficacy of susceptibility-guided therapy via gastric juice PCR for the first-line Hp eradication. In Asia-Pacific area, where standard triple therapy is still adopted by the majority of the physicians, it is a recommended alternative to overcome the increasing antibiotic resistance.
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Affiliation(s)
- Meng-Shu Hsieh
- Department of Internal Medicine, Kaohsiung Municipal Siaogang Hospital, Kaohsiung, Taiwan
| | - Fu-Chen Kuo
- School of Medicine, College of Medicine, E-Da Hospital, I-Shou University, Kaohsiung, Taiwan
| | - Meng-Chieh Wu
- Department of Internal Medicine, Kaohsiung Municipal Ta-Tung Hospital, Kaohsiung, Taiwan
| | - Jiunn-Wei Wang
- Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan; Department of Medicine, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan
| | - Chung-Jung Liu
- Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan
| | - Neng-Sheng Chu
- Department of Internal Medicine, Kaohsiung Municipal Siaogang Hospital, Kaohsiung, Taiwan
| | - Pei-Yun Tsai
- Department of Nursing, Kaohsiung Municipal Siaogang Hospital, Kaohsiung, Taiwan
| | - Ping-I Hsu
- Department of Medicine, An Nan Hospital, China Medical University, Tainan, Taiwan
| | - I-Chen Wu
- Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan; Department of Medicine, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan
| | - Jeng-Yih Wu
- Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan; Department of Medicine, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan
| | - Deng-Chyang Wu
- Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan; Department of Medicine, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan
| | - Fang-Jung Yu
- Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan; Department of Medicine, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
| | - Chao-Hung Kuo
- Department of Internal Medicine, Kaohsiung Municipal Siaogang Hospital, Kaohsiung, Taiwan; Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan; Department of Medicine, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
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Crowley E, Hussey S. Helicobacter pylori in Childhood. PEDIATRIC GASTROINTESTINAL AND LIVER DISEASE 2021:275-292.e12. [DOI: 10.1016/b978-0-323-67293-1.00027-x] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/04/2025]
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Szymczak A, Ferenc S, Majewska J, Miernikiewicz P, Gnus J, Witkiewicz W, Dąbrowska K. Application of 16S rRNA gene sequencing in Helicobacter pylori detection. PeerJ 2020; 8:e9099. [PMID: 32440373 PMCID: PMC7229771 DOI: 10.7717/peerj.9099] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/29/2019] [Accepted: 04/09/2020] [Indexed: 12/13/2022] Open
Abstract
Helicobacter pylori is one of the major stomach microbiome components, promoting development of inflammation and gastric cancer in humans. H. pylori has a unique ability to transform into a coccoidal form which is difficult to detect by many diagnostic methods, such as urease activity detection, and even histopathological examination. Here we present a comparison of three methods for H. pylori identification: histological assessment (with eosin, hematoxylin, and Giemsa staining), polymerase chain reaction (PCR) detection of urease (ureA specific primers), and detection by 16S rRNA gene sequencing. The study employed biopsies from the antral part of the stomach (N = 40). All samples were assessed histologically which revealed H. pylori in eight patients. Bacterial DNA isolated from the bioptates was used as a template for PCR reaction and 16S rRNA gene sequencing that revealed H. pylori in 13 and in 20 patients, respectively. Thus, 16S rRNA gene sequencing was the most sensitive method for detection of H. pylori in stomach biopsy samples.
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Affiliation(s)
- Aleksander Szymczak
- Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland
| | - Stanisław Ferenc
- Regional Specialist Hospital in Wrocław, Research and Development Center, Wrocław, Poland
| | - Joanna Majewska
- Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland
| | - Paulina Miernikiewicz
- Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland
| | - Jan Gnus
- Medical Academy in Wroclaw, Wrocław, Poland
| | - Wojciech Witkiewicz
- Regional Specialist Hospital in Wrocław, Research and Development Center, Wrocław, Poland
| | - Krystyna Dąbrowska
- Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland
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Prevalence of Helicobacter pylori Infection and Diagnostic Methods in the Middle East and North Africa Region. ACTA ACUST UNITED AC 2020; 56:medicina56040169. [PMID: 32283700 PMCID: PMC7230269 DOI: 10.3390/medicina56040169] [Citation(s) in RCA: 15] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/25/2020] [Revised: 04/05/2020] [Accepted: 04/06/2020] [Indexed: 12/15/2022]
Abstract
Background and Objectives:Helicobacter pylori (H. pylori) infection is common worldwide and may cause gastroduodenal complications, including cancer. In this review, we examine the prevalence and distribution of various H. pylori genotypes and the risk factors for H. pylori infection, particularly in the Middle East and North Africa (MENA) region. We also introduce different global screening methods and guidelines and compare them to those currently in use in the MENA region. Materials and Methods: We searched the Google Scholar, PubMed, and Saudi Digital Library (SDL) databases for clinical trials and articles published in English. The data collection was mainly focused on MENA countries. However, for H. pylori genotypes and diagnostic methods, studies conducted in other regions or reporting global practices and guidelines were also included to allow a comparison with those in the MENA region. We also included studies examining the prevalence of H. pylori infection in healthy participants. Results:H. pylori infection is highly prevalent in the MENA region, mainly because of the accumulation of risk factors in developing countries. Herein, we highlight a lack of good quality studies on the prevalence of various H. pylori genotypes in the MENA region as well as a need for standard diagnostic methods and screening guidelines. Due to the complications associated with H. pylori, we recommend routine screening for H. pylori infection in all gastroenterology patients admitted in the MENA region. Conclusion: Concerted effort will first be required to validate affordable, non-invasive, and accurate diagnostic methods and to establish local guidelines with adapted cut-off values for the interpretation of the test results.
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Sabbagh P, Mohammadnia-Afrouzi M, Javanian M, Babazadeh A, Koppolu V, Vasigala VR, Nouri HR, Ebrahimpour S. Diagnostic methods for Helicobacter pylori infection: ideals, options, and limitations. Eur J Clin Microbiol Infect Dis 2018; 38:55-66. [PMID: 30414090 DOI: 10.1007/s10096-018-3414-4] [Citation(s) in RCA: 92] [Impact Index Per Article: 13.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/10/2018] [Accepted: 10/26/2018] [Indexed: 12/13/2022]
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Genotyping and antimicrobial resistance patterns of Helicobacter pylori in human and dogs associated with A2142G and A2143G point mutations in clarithromycin resistance. Microb Pathog 2018; 123:330-338. [PMID: 30031039 DOI: 10.1016/j.micpath.2018.07.016] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/15/2018] [Revised: 07/12/2018] [Accepted: 07/13/2018] [Indexed: 02/07/2023]
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Guzman J, Téné N, Touchard A, Castillo D, Belkhelfa H, Haddioui-Hbabi L, Treilhou M, Sauvain M. Anti-Helicobacter pylori Properties of the Ant-Venom Peptide Bicarinalin. Toxins (Basel) 2017; 10:E21. [PMID: 29286296 PMCID: PMC5793108 DOI: 10.3390/toxins10010021] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/06/2017] [Revised: 12/22/2017] [Accepted: 12/23/2017] [Indexed: 12/25/2022] Open
Abstract
The venom peptide bicarinalin, previously isolated from the ant Tetramorium bicarinatum, is an antimicrobial agent with a broad spectrum of activity. In this study, we investigate the potential of bicarinalin as a novel agent against Helicobacter pylori, which causes several gastric diseases. First, the effects of synthetic bicarinalin have been tested against Helicobacter pylori: one ATCC strain, and forty-four isolated from stomach ulcer biopsies of Peruvian patients. Then the cytoxicity of bicarinalin on human gastric cells and murine peritoneal macrophages was measured using XTT and MTT assays, respectively. Finally, the preventive effect of bicarinalin was evaluated by scanning electron microscopy using an adherence assay of H. pylori on human gastric cells treated with bicarinalin. This peptide has a potent antibacterial activity at the same magnitude as four antibiotics currently used in therapies against H. pylori. Bicarinalin also inhibited adherence of H. pylori to gastric cells with an IC50 of 0.12 μg·mL-1 and had low toxicity for human cells. Scanning electron microscopy confirmed that bicarinalin can significantly decrease the density of H. pylori on gastric cells. We conclude that Bicarinalin is a promising compound for the development of a novel and effective anti-H. pylori agent for both curative and preventive use.
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Affiliation(s)
- Jesus Guzman
- Laboratorios de Investigación y Desarrollo, Universidad Peruana Cayetano Heredia (UPCH), Lima 34, Peru.
| | - Nathan Téné
- EA7417-BTSB, Université Fédérale Toulouse Midi-Pyrénées, INU Champollion, 81012 Albi, France.
| | - Axel Touchard
- EA7417-BTSB, Université Fédérale Toulouse Midi-Pyrénées, INU Champollion, 81012 Albi, France.
| | - Denis Castillo
- Laboratorios de Investigación y Desarrollo, Universidad Peruana Cayetano Heredia (UPCH), Lima 34, Peru.
| | - Haouaria Belkhelfa
- Fonderephar, Université Fédérale Toulouse Midi-Pyrénées, Faculté des Sciences Pharmaceutiques, 31062 Toulouse, France.
| | - Laila Haddioui-Hbabi
- Fonderephar, Université Fédérale Toulouse Midi-Pyrénées, Faculté des Sciences Pharmaceutiques, 31062 Toulouse, France.
| | - Michel Treilhou
- EA7417-BTSB, Université Fédérale Toulouse Midi-Pyrénées, INU Champollion, 81012 Albi, France.
| | - Michel Sauvain
- Laboratorios de Investigación y Desarrollo, Universidad Peruana Cayetano Heredia (UPCH), Lima 34, Peru.
- UMR 152 PHARMADEV, Université Fédérale Toulouse Midi-Pyrénées, IRD, 31062 Toulouse, France.
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Alebie G. Prevalence of Helicobacter Pylori Infection and Associated Factors among Gastritis Students in Jigjiga University, Jigjiga, Somali Regional State of Ethiopia. ACTA ACUST UNITED AC 2016. [DOI: 10.15406/jbmoa.2016.03.00060] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/17/2022]
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Wang YK, Kuo FC, Liu CJ, Wu MC, Shih HY, Wang SSW, Wu JY, Kuo CH, Huang YK, Wu DC. Diagnosis of Helicobacter pylori infection: Current options and developments. World J Gastroenterol 2016. [PMID: 26523098 DOI: 10.3748/wjg.v21.i40.11221.] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 10/11/2022] Open
Abstract
Accurate diagnosis of Helicobacter pylori (H. pylori) infection is a crucial part in the effective management of many gastroduodenal diseases. Several invasive and non-invasive diagnostic tests are available for the detection of H. pylori and each test has its usefulness and limitations in different clinical situations. Although none can be considered as a single gold standard in clinical practice, several techniques have been developed to give the more reliable results. Invasive tests are performed via endoscopic biopsy specimens and these tests include histology, culture, rapid urease test as well as molecular methods. Developments of endoscopic equipment also contribute to the real-time diagnosis of H. pylori during endoscopy. Urea breathing test and stool antigen test are most widely used non-invasive tests, whereas serology is useful in screening and epidemiological studies. Molecular methods have been used in variable specimens other than gastric mucosa. More than detection of H. pylori infection, several tests are introduced into the evaluation of virulence factors and antibiotic sensitivity of H. pylori, as well as screening precancerous lesions and gastric cancer. The aim of this article is to review the current options and novel developments of diagnostic tests and their applications in different clinical conditions or for specific purposes.
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Affiliation(s)
- Yao-Kuang Wang
- Yao-Kuang Wang, Department of Internal Medicine, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung 812, Taiwan
| | - Fu-Chen Kuo
- Yao-Kuang Wang, Department of Internal Medicine, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung 812, Taiwan
| | - Chung-Jung Liu
- Yao-Kuang Wang, Department of Internal Medicine, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung 812, Taiwan
| | - Meng-Chieh Wu
- Yao-Kuang Wang, Department of Internal Medicine, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung 812, Taiwan
| | - Hsiang-Yao Shih
- Yao-Kuang Wang, Department of Internal Medicine, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung 812, Taiwan
| | - Sophie S W Wang
- Yao-Kuang Wang, Department of Internal Medicine, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung 812, Taiwan
| | - Jeng-Yih Wu
- Yao-Kuang Wang, Department of Internal Medicine, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung 812, Taiwan
| | - Chao-Hung Kuo
- Yao-Kuang Wang, Department of Internal Medicine, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung 812, Taiwan
| | - Yao-Kang Huang
- Yao-Kuang Wang, Department of Internal Medicine, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung 812, Taiwan
| | - Deng-Chyang Wu
- Yao-Kuang Wang, Department of Internal Medicine, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung 812, Taiwan
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Kronsteiner B, Bassaganya-Riera J, Philipson C, Viladomiu M, Carbo A, Abedi V, Hontecillas R. Systems-wide analyses of mucosal immune responses to Helicobacter pylori at the interface between pathogenicity and symbiosis. Gut Microbes 2016; 7:3-21. [PMID: 26939848 PMCID: PMC4856448 DOI: 10.1080/19490976.2015.1116673] [Citation(s) in RCA: 32] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 09/16/2015] [Revised: 10/29/2015] [Accepted: 10/31/2015] [Indexed: 02/08/2023] Open
Abstract
Helicobacter pylori is the dominant member of the gastric microbiota in over half of the human population of which 5-15% develop gastritis or gastric malignancies. Immune responses to H. pylori are characterized by mixed T helper cell, cytotoxic T cell and NK cell responses. The presence of Tregs is essential for the control of gastritis and together with regulatory CX3CR1+ mononuclear phagocytes and immune-evasion strategies they enable life-long persistence of H. pylori. This H. pylori-induced regulatory environment might contribute to its cross-protective effect in inflammatory bowel disease and obesity. Here we review host-microbe interactions, the development of pro- and anti-inflammatory immune responses and how the latter contribute to H. pylori's role as beneficial member of the gut microbiota. Furthermore, we present the integration of existing and new data into a computational/mathematical model and its use for the investigation of immunological mechanisms underlying initiation, progression and outcomes of H. pylori infection.
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Affiliation(s)
- Barbara Kronsteiner
- Nutritional Immunology and Molecular Medicine Laboratory and Center for Modeling Immunity to Enteric Pathogens; Virginia Bioinformatics Institute; Virginia Tech; Blacksburg, VA, USA
| | - Josep Bassaganya-Riera
- Nutritional Immunology and Molecular Medicine Laboratory and Center for Modeling Immunity to Enteric Pathogens; Virginia Bioinformatics Institute; Virginia Tech; Blacksburg, VA, USA
| | | | - Monica Viladomiu
- Nutritional Immunology and Molecular Medicine Laboratory and Center for Modeling Immunity to Enteric Pathogens; Virginia Bioinformatics Institute; Virginia Tech; Blacksburg, VA, USA
| | | | - Vida Abedi
- Nutritional Immunology and Molecular Medicine Laboratory and Center for Modeling Immunity to Enteric Pathogens; Virginia Bioinformatics Institute; Virginia Tech; Blacksburg, VA, USA
| | - Raquel Hontecillas
- Nutritional Immunology and Molecular Medicine Laboratory and Center for Modeling Immunity to Enteric Pathogens; Virginia Bioinformatics Institute; Virginia Tech; Blacksburg, VA, USA
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Kalali B, Formichella L, Gerhard M. Diagnosis of Helicobacter pylori: Changes towards the Future. Diseases 2015; 3:122-135. [PMID: 28943614 PMCID: PMC5548244 DOI: 10.3390/diseases3030122] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/12/2015] [Revised: 06/18/2015] [Accepted: 06/19/2015] [Indexed: 12/19/2022] Open
Abstract
Since the first evidence demonstrating the dramatically high incidence of H. pylori infection and the subsequent medical challenges it incurs, health management of H. pylori infection has been a high priority for health authorities worldwide. Despite a decreasing rate of infection in western countries, prevalence of H. pylori infection in developing and in some industrial countries is still very high. Whereas treatment and vaccination against H. pylori is a contemporary issue in medical communities, selective treatment and prior high-throughput screening of the subject population is a major concern of health organizations. So far, diagnostic tests are either elaborative and require relatively advanced medical care infrastructure or they do not fulfill the criteria recommended by the Maastricht IV/Florence consensus report. In this review, in light of recent scientific studies, we highlight current and possible future approaches for the diagnosis of H. pylori. We point out that novel non-invasive tests may not only cover the requirements of gold standard methods in H. pylori detection but also offer the potential for risk stratification of infection in a high throughput manner.
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Affiliation(s)
- Behnam Kalali
- Institute of Medical Microbiology, Immunology and Hygiene, TU-Munich, Troger Str.30, 81675 Munich, Germany.
- ImevaX GmbH, Grillparzer Str.18, 81675 Munich, Germany.
| | - Luca Formichella
- Institute of Medical Microbiology, Immunology and Hygiene, TU-Munich, Troger Str.30, 81675 Munich, Germany.
| | - Markus Gerhard
- Institute of Medical Microbiology, Immunology and Hygiene, TU-Munich, Troger Str.30, 81675 Munich, Germany.
- ImevaX GmbH, Grillparzer Str.18, 81675 Munich, Germany.
- German Center for Infection Research (DZIF), Partner site Munich, Troger Str.30, 81675 Munich, Germany.
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Siavoshi F, Saniee P, Khalili-Samani S, Hosseini F, Malakutikhah F, Mamivand M, Shahreza S, Sharifi AH. Evaluation of methods for H. pylori detection in PPI consumption using culture, rapid urease test and smear examination. ANNALS OF TRANSLATIONAL MEDICINE 2015; 3:11. [PMID: 25705643 DOI: 10.3978/j.issn.2305-5839.2014.11.16] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Key Words] [Subscribe] [Scholar Register] [Received: 08/24/2014] [Accepted: 10/29/2014] [Indexed: 12/23/2022]
Abstract
BACKGROUND Culture, rapid urease test (RUT) and smear examination have been used as reliable methods for diagnosis of H. pylori infection. Accurate performance of these tests requires good quality biopsies with considerable number of bacterial cells. However, consumption of proton pump inhibitors (PPIs) affects growth and urease activity of H. pylori, leading to false negative results. In this study the efficacy of culture, RUT and smear examination was assessed and the effect of PPI consumption was evaluated. METHODS Two antral biopsies from 530 dyspeptic patients with and without PPI consumption were used for RUT, culture and smear examination. Statistical analysis was used to determine the association between results of culture, RUT or smear examination and PPI consumption. Sensitivity and specificity of three tests were calculated by standard methods. RESULTS H. pylori infection was detected in 40% of patients by culture, 48.3% by RUT and 21.1% by smear examination and the overall detection rate was 54%. A strong correlation was found between PPI consumption and negative results of culture and RUT (P<0.05) but not smear examination. The sensitivity of RUT was reduced as a result of PPI consumption. This reduction was more profound in 1-hr RUT (92.2% to 74.4%) compared with 24-hr RUT (93.9% to 81.6%). CONCLUSIONS Prevalence of H. pylori was declined, compared with previous studies. This decrement could be due to false negative results of H. pylori diagnostic tests, among which culture and RUT are mostly affected by PPI. Accordingly, PPI consumption should be stopped before performance of endoscopy.
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Affiliation(s)
- Farideh Siavoshi
- 1 Department of Microbiology, School of Biology, University College of Sciences, University of Tehran, Tehran 14176-14411, Iran ; 2 Digestive Disease Research Institute, Tehran University of Medical Sciences, Tehran 14117-13135, Iran
| | - Parastoo Saniee
- 1 Department of Microbiology, School of Biology, University College of Sciences, University of Tehran, Tehran 14176-14411, Iran ; 2 Digestive Disease Research Institute, Tehran University of Medical Sciences, Tehran 14117-13135, Iran
| | - Saman Khalili-Samani
- 1 Department of Microbiology, School of Biology, University College of Sciences, University of Tehran, Tehran 14176-14411, Iran ; 2 Digestive Disease Research Institute, Tehran University of Medical Sciences, Tehran 14117-13135, Iran
| | - Farideh Hosseini
- 1 Department of Microbiology, School of Biology, University College of Sciences, University of Tehran, Tehran 14176-14411, Iran ; 2 Digestive Disease Research Institute, Tehran University of Medical Sciences, Tehran 14117-13135, Iran
| | - Fahimeh Malakutikhah
- 1 Department of Microbiology, School of Biology, University College of Sciences, University of Tehran, Tehran 14176-14411, Iran ; 2 Digestive Disease Research Institute, Tehran University of Medical Sciences, Tehran 14117-13135, Iran
| | - Marzieh Mamivand
- 1 Department of Microbiology, School of Biology, University College of Sciences, University of Tehran, Tehran 14176-14411, Iran ; 2 Digestive Disease Research Institute, Tehran University of Medical Sciences, Tehran 14117-13135, Iran
| | - Somayeh Shahreza
- 1 Department of Microbiology, School of Biology, University College of Sciences, University of Tehran, Tehran 14176-14411, Iran ; 2 Digestive Disease Research Institute, Tehran University of Medical Sciences, Tehran 14117-13135, Iran
| | - Amir Houshang Sharifi
- 1 Department of Microbiology, School of Biology, University College of Sciences, University of Tehran, Tehran 14176-14411, Iran ; 2 Digestive Disease Research Institute, Tehran University of Medical Sciences, Tehran 14117-13135, Iran
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Current Paradigm and Future Directions for Treatment of Helicobacter pylori Infection. ACTA ACUST UNITED AC 2014; 12:373-84. [PMID: 25187235 DOI: 10.1007/s11938-014-0027-6] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/07/2023]
Abstract
OPINION STATEMENT Once easily eradicated with triple or quadruple therapy, Helicobacter pylori infection has become increasingly resistant to traditional first-line treatment regimens because of emerging antibiotic resistance coupled with poor patient compliance with completing the treatment course. Given decreasing H. pylori eradication rates, there is considerable interest in evaluating new antibiotic combinations and regimens, the addition of probiotics, and the development of new paradigms such as concomitant, sequential, and hybrid medication dosing strategies. Unfortunately, efforts thus far have not universally improved treatment responses, as promising early results were often not extrapolated to wider populations. This is probably due largely to regional variation in H. pylori resistance patterns. Ideally, the standard of care should be dictated by knowledge of local H. pylori antimicrobial resistance patterns and clinical success rates rather than by empiric extrapolation from the literature. Unfortunately, such knowledge is usually lacking in the USA. The expectation of a first-line regimen is a minimum 80 % eradication rate in the local population. Standard triple therapy with a proton pump inhibitor (PPI), amoxicillin, and clarithromycin may still be effective in some areas; however, in populations with high clarithromycin resistance, quadruple therapy with a metronidazole-based regimen may be a better choice, and concomitant, sequential, or hybrid dosing schedules should also be considered as possible first-line choices. Second- and third-line treatment regimens consist of levofloxacin-based and rifabutin-based therapies, respectively. Further work should be directed at establishing local resistance patterns and eradication rates, developing H. pylori-specific antibiotics, and starting culture-guided treatment programs. Ultimately, the development of an H. pylori vaccine would bypass any issues with antibiotic resistance by preventing the acquisition of infection altogether.
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Abstract
We developed a new transport medium (GESA--Helicobacter pylori transport medium [publication no. WO/2014/019696, patent pending no. PCT/EP2013/002292; Liofilchem s.r.l., Roseto degli Abruzzi, Teramo, Italy]) for recovery of Helicobacter pylori from gastric biopsy samples. GESA transport medium, in a semisolid state, provides the optimal conditions for maintaining the viability of the microorganism over time. The efficacy of the transport medium was assessed through in vitro and ex vivo experiments. We were able to recover different suspensions of H. pylori ATCC 43629 and H. pylori 13 A in GESA transport medium stored at 4 °C for up to 10 days. In particular, with a starting inoculum of ∼ 10(5) CFU, after 7 days of storage, 150 ± 25 CFU and 40 ± 7 CFU of the reference and clinical strains were detected, respectively. H. pylori colonies were isolated from gastric specimens taken from both the antrum and the fundus in 68 (90.66%) of 75 urea breath test (UBT)-positive patients. Moreover, GESA transport medium allowed the recovery and isolation of H. pylori colonies from additional biopsy samples from 13 of the 75 detected subjects at up to 10 days of biopsy sample storage at 4 °C. Finally, GESA transport medium preserved its characteristics when stored at 4°C for 1 year from its preparation, thus allowing good recovery of H. pylori. GESA transport medium can be considered a standardized transport medium with high performance that optimizes the recovery rate of H. pylori grown by culture.
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Lopes AI, Vale FF, Oleastro M. Helicobacter pylori infection - recent developments in diagnosis. World J Gastroenterol 2014; 20:9299-9313. [PMID: 25071324 PMCID: PMC4110561 DOI: 10.3748/wjg.v20.i28.9299] [Citation(s) in RCA: 51] [Impact Index Per Article: 4.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/08/2013] [Accepted: 04/16/2014] [Indexed: 02/06/2023] Open
Abstract
Considering the recommended indications for Helicobacter pylori (H. pylori) eradication therapy and the broad spectrum of available diagnostic methods, a reliable diagnosis is mandatory both before and after eradication therapy. Only highly accurate tests should be used in clinical practice, and the sensitivity and specificity of an adequate test should exceed 90%. The choice of tests should take into account clinical circumstances, the likelihood ratio of positive and negative tests, the cost-effectiveness of the testing strategy and the availability of the tests. This review concerns some of the most recent developments in diagnostic methods of H. pylori infection, namely the contribution of novel endoscopic evaluation methodologies for the diagnosis of H. pylori infection, such as magnifying endoscopy techniques and chromoendoscopy. In addition, the diagnostic contribution of histology and the urea breath test was explored recently in specific clinical settings and patient groups. Recent studies recommend enhancing the number of biopsy fragments for the rapid urease test. Bacterial culture from the gastric biopsy is the gold standard technique, and is recommended for antibiotic susceptibility test. Serology is used for initial screening and the stool antigen test is particularly used when the urea breath test is not available, while molecular methods have gained attention mostly for detecting antibiotic resistance.
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Ramis IB, de Moraes EP, Fernandes MS, Mendoza-Sassi R, Rodrigues O, Juliano CRV, Scaini CJ, da Silva PEA. Evaluation of diagnostic methods for the detection of Helicobacter pylori in gastric biopsy specimens of dyspeptic patients. Braz J Microbiol 2012; 43:903-8. [PMID: 24031905 PMCID: PMC3768872 DOI: 10.1590/s1517-83822012000300008] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/08/2011] [Accepted: 06/07/2012] [Indexed: 12/15/2022] Open
Abstract
Helicobacter pylori infects nearly 50% of the world’s population. This microorganism is accepted as the most important agent of gastritis and as a risk factor for peptic ulcer disease and gastric adenocarcinoma. Currently many diagnostic methods exist for detecting H. pylori, however they all have limitations, thus it is recommend a combination of at least two methods. The aim of this study was to evaluate diagnostic methods, such as in-house urease test, culture and Polymerase Chain Reaction (PCR), for the detection of the H. pylori in gastric biopsy specimens of 144 dyspeptic patients, using as gold standard the association between histology and rapid urease test. According to the gold standard used in this study, 48 (33.3%) patients were infected with H. pylori, while 96 (66.7%) were classified as not infected. The in-house urease test and the PCR were the most sensitive methods (100%), followed by culture (85.4%). However, the inhouse urease test and the culture were the most specific (100%), followed by PCR (75%). In conclusion, this study showed that, in comparison with the combination of histology and rapid urease test, the in-house urease test and the PCR presented 100% of sensitivity in the diagnosis of gastric infection by H. pylori, while the in-house urease test and the culture reached 100% of specificity. These finding suggest that the combination of two or more methods may improve the accuracy of the H. pylori detection.
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Affiliation(s)
- Ivy Bastos Ramis
- Laboratório de Biologia Molecular, Universidade Federal do Rio Grande, Rio Grande , Rio Grande do Sul , Brasil
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Abstract
Over the past 25 years, a variety of methods have been developed for culture of Helicobacter pylori in vitro. H. pylori is a capnophilic and microaerophilic organism that is typically cultured using complex culture media. Analysis of H. pylori growth in chemically defined media has provided insight into the nutritional requirements, physiology, and metabolic capacities of this organism.
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Patient factors affecting culture of Helicobacter pylori isolated from gastric mucosal specimens. Adv Med Sci 2011; 55:161-6. [PMID: 20639184 DOI: 10.2478/v10039-010-0028-1] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/07/2023]
Abstract
PURPOSE Culture is one of the methods used for detecting Helicobacter pylori in the stomach. However, since it is costly, labor-consuming, and in a number of infected subjects gives a false negative result, the procedure is not routinely used. The aim of the study was to analyze some of the factors that may affect the outcome of H. pylori culture from endoscopic gastric mucosal specimens. MATERIAL AND METHODS The study was conducted in a group of 265 subjects. The culture of gastric mucosal specimens was verified by urease test and histological examination. If the culture result was not consistent with one or two verifying tests, an additional two tests were used, i.e. H. pylori antigens in stool samples and anti-H. pylori antibodies in blood serum. RESULTS In patients infected with H. pylori (at least two positive diagnostic tests), the analysis of factors that may affect the culture outcome revealed that neither age, gender, smoking, history of eradication, endoscopic diagnosis, use of proton pump inhibitors, ultrasonography of the abdomen or chest radiology performed the day before or on the day of gastroscopy, nor preparation for colonoscopy using osmotic fluids 1-2 days prior to gastroscopy had an effect on the culture outcome. Only high activity of gastritis (neutrophil infiltration) and low bacterial load in gastric mucosal specimens as well as drinking alcohol and the use of histamine H₂ receptor blockers reduced culture efficacy in infected subjects. CONCLUSIONS High activity of gastritis, low bacterial load, drinking alcohol and the use of histamine H₂ receptor blockers can be the cause of failed H. pylori culture from gastric mucosa in the infected subjects. These factors should be taken into consideration when qualifying patients for the test and interpreting the results.
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Hussey S, Jones NL. Helicobacter pylori in Childhood. PEDIATRIC GASTROINTESTINAL AND LIVER DISEASE 2011:293-308.e10. [DOI: 10.1016/b978-1-4377-0774-8.10028-4] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/03/2025]
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Radić M, Kaliterna DM, Radić J. Helicobacter pylori infection and systemic sclerosis-is there a link? Joint Bone Spine 2010; 78:337-40. [PMID: 21145276 DOI: 10.1016/j.jbspin.2010.10.005] [Citation(s) in RCA: 21] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/13/2010] [Accepted: 10/08/2010] [Indexed: 12/11/2022]
Abstract
Over the last 20 years, increasing evidence has accumulated to implicate infectious agents in the etiology of systemic sclerosis (SSc) and Raynaud's phenomenon. Infection rates in patients with SSc compared with those in control populations do not provide clear support for any specific pathogen. However, increased antibody titers, a preponderance of specific strains in patients with SSc, and evidence of molecular mimicry inducing autoimmune responses suggest mechanisms by which infectious agents may contribute to the development and progression of SSc. Helicobacter pylori (H. pylori) has been associated with diseases such as autoimmune gastritis, Sjögren's syndrome, atherosclerosis, immune thrombocytopenia purpura, inflammatory bowel diseases and autoimmune pancreatitis, in each of which it seems to play a pathogenetic, but it has also been suggested that it may help to protect against the development of autoimmune gastritis, multiple sclerosis, systemic lupus erythemathosus and inflammatory bowel diseases. A systematic literature search was carried out in MEDLINE, EMBASE, Cochrane Library and ACR/EULAR meeting abstracts. We hypotheses that H. pylori infection might play a critical role in the pathogenesis of SSc. Here we review studies examining the potential involvement of H. pylori infection in SSc.
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Affiliation(s)
- Mislav Radić
- Department of Rheumatology, University Hospital, Split, Croatia.
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Detection of Helicobacter pylori by phenotypic and genotypic methods. Dig Dis Sci 2010; 55:1643-8. [PMID: 19693671 DOI: 10.1007/s10620-009-0928-8] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 12/19/2008] [Accepted: 07/16/2009] [Indexed: 12/16/2022]
Abstract
AIMS This research evaluated the utilization of a urease in-house test, culture and molecular method (ureA PCR) as a diagnostic tool for Helicobacter pylori infection. Furthermore, we assessed the presence of the cagA gene in the specimens and in isolated strains that were positive for ureA by PCR positive. RESULTS Sensitivity and specificity, respectively, were 100 and 95.8% for the urease in-house test 93.3 and 95.8 for the ureA PCR assay of the specimen and 100 and 100% for the culture. The presence of the cagA gene was observed in eight (53%) ureA-positive samples. CONCLUSIONS In this study, we found that the PCR technique has applicability in the study of cagA, and other genes related to the H. pylori pathogen. This method can be applied to samples directly from biopsy or isolated from the bacteria.
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Tanih NF, Dube C, Green E, Mkwetshana N, Clarke AM, Ndip LM, Ndip RN. An African perspective on Helicobacter pylori: prevalence of human infection, drug resistance, and alternative approaches to treatment. ANNALS OF TROPICAL MEDICINE AND PARASITOLOGY 2010; 103:189-204. [PMID: 19341534 DOI: 10.1179/136485909x398311] [Citation(s) in RCA: 21] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/31/2022]
Abstract
Helicobacter pylori is a Gram-negative, micro-aerophilic, motile, curved rod that inhabits the gastric mucosa of the human stomach. It chronically infects thousands of millions of people world-wide, and is one of the most genetically diverse of bacterial species. Infection with the bacterium leads to chronic gastritis, peptic ulceration, gastric cancers and gastric mucosa-associated lymphoid-tissue (MALT) lymphoma. The prevalence of infection appears to be partly determined by geographical and socio-demographic factors, being higher in Africa than elsewhere. Current treatment, based on potent combinations that each consist of a proton-pump inhibitor and two antibiotics, is successful in 80%-90% of patients. Some undesirable side-effects, poor patient compliance and drug resistance are, however, associated with significant levels of treatment failure and with contra-indications for some patients. Antibiotic resistance in H. pylori is a growing global concern that merits the urgent attention of public-health authorities. Numerous pieces of clinical evidence have revealed that eradication of the organism from a patient results in improvement of gastritis and drastically decreases the frequency of relapse of gastric and duodenal ulcers. Natural products, including medicinal plants and honey, may offer useful alternatives in the treatment of H. pylori-related infections.
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Affiliation(s)
- N F Tanih
- Microbial Pathogenicity and Molecular Epidemiology Research Group, Department of Biochemistry and Microbiology, Faculty of Science and Agriculture, University of Fort Hare, P.M.B. X1314, Alice 5700, South Africa
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Brgers R, Schneider-Brachert W, Reischl U, Behr A, Hiller KA, Lehn N, Schmalz G, Ruhl S. Helicobacter pyloriin human oral cavity and stomach. Eur J Oral Sci 2008; 116:297-304. [DOI: 10.1111/j.1600-0722.2008.00543.x] [Citation(s) in RCA: 56] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
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Ndip RN, Malange Takang AE, Ojongokpoko JEA, Luma HN, Malongue A, Akoachere JFTK, Ndip LM, MacMillan M, Weaver LT. Helicobacter pylori isolates recovered from gastric biopsies of patients with gastro-duodenal pathologies in Cameroon: current status of antibiogram. Trop Med Int Health 2008; 13:848-54. [PMID: 18384477 DOI: 10.1111/j.1365-3156.2008.02062.x] [Citation(s) in RCA: 37] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
OBJECTIVE To determine the prevalence of Helicobacter pylori in patients with gastro-duodenal pathologies and the susceptibility patterns of isolates to the currently recommended antibiotic treatment regimen used in Cameroon. METHODS Consecutive dyspeptic patients referred to Douala General Hospital, Cameroon for endoscopy were recruited in the study. Gastric biopsies were collected from the patients and H. pylori isolated and identified following standard microbiology and biochemical techniques. Antibiotic susceptibility was determined by disk diffusion and agar dilution methods against clarithromycin, tetracycline, amoxicillin and metronidazole. Data were analysed using chi-square test and significance considered at P < 0.05. RESULTS Seventy-one (92.2%) of the 77 patients (mean age 44.5 +/- 15.7, range 15-77 years) were positive for H. pylori. The antibiotic susceptibility rates were 56% for tetracycline, 55.3% for clarithromycin, 14.4% for amoxicillin and 6.8% for metronidazole. The prevalence of clarithromycin resistance in males vs. females was 42.1%vs. 46.7%, while for metronidazole it was 89.5%vs. 94.7% (P > 0.05). Antimicrobial susceptibility results also revealed 12 antibiotypes based on resistance to the antimicrobial agents investigated. The resistance pattern, amoxicillin and metronidazole (AMR(R) MET(R)) was the most common (23.7%) amongst the isolates. More than 60% of the isolates exhibited multi-drug resistance to three or four antibiotics. CONCLUSION Multi-drug resistance is common against the current treatment regimen in Cameroon and, therefore, calls for urgent studies involving newer and broad spectrum antibiotics to address the problem.
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Affiliation(s)
- Roland N Ndip
- Department of Biochemistry and Microbiology, Faculty of Science, University of Buea, Buea, Cameroon.
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Ndip RN, Malange Tarkang AE, Mbullah SM, Luma HN, Malongue A, Ndip LM, Nyongbela K, Wirmum C, Efange SMN. In vitro anti-Helicobacter pylori activity of extracts of selected medicinal plants from North West Cameroon. JOURNAL OF ETHNOPHARMACOLOGY 2007; 114:452-7. [PMID: 17913416 DOI: 10.1016/j.jep.2007.08.037] [Citation(s) in RCA: 56] [Impact Index Per Article: 3.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/27/2007] [Revised: 07/30/2007] [Accepted: 08/22/2007] [Indexed: 05/17/2023]
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE Helicobacter pylori, a gram negative microaerophilic bacterium is a major etiological agent in duodenal, peptic and gastric ulcers. The growing problem of antibiotic resistance by the organism demands the search for novel compounds from plant based sources. AIM OF STUDY The present study is aimed at evaluating the antimicrobial activity of some selected medicinal plants on clinical isolates of H. pylori circulating in Cameroon in a bid to identify potential sources of cheap starting materials for the synthesis of new drugs. MATERIALS AND METHODS Gastric biopsy samples were obtained from patients presenting with gastroduodenal complications. H. pylori was isolated from the specimens following standard microbiology procedures. The disk diffusion method was used to determine the susceptibility of 15 isolates to ten methanol plant extracts (Ageratum conyzoides, Scleria striatinux, Lycopodium cernua, Acanthus montanus, Eryngium foetidium, Aulutandria kamerunensis, Tapeinachilus ananassae, Euphorbia hirta, Emilia coccinea and Scleria verrucosa). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) for the most active plant extracts were also determined by the agar dilution method. Results were analyzed statistically by the Fisher's exact test. RESULTS All the plants tested demonstrated antimicrobial activity with zone diameters of inhibition ranging from 0-30mm. Of these, A. conyzoides, S. striatinux and L. cernua showed very potent antibacterial activity on the isolates. The lowest MIC and MBC recorded were 0.032mg/mL and 0.098mg/mL respectively. However, the MIC of the extracts ranged from 0.032-1.0mg/mL for S. striatinux; 0.063-0.5mg/mL for L. cernua and 0.063-1.0mg/mL for A. conyzoides. The MBC of the extracts ranged from 0.098-15.0mg/mL for S. striatinux; 0.098-12.5mg/mL for A. conyzoides, and 0.195-12.5mg/mL for L. cernua. The extracts had a wide spectrum of activity. The three most potent extracts possessed significant (P<0.05) inhibitory activities. CONCLUSION The plant extracts may contain compounds with therapeutic activity.
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Affiliation(s)
- Roland N Ndip
- Department of Biochemistry and Microbiology, University of Buea, Cameroon.
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Abstract
Since the first successful attempts of growing Helicobacter pylori, a variety of tests for detecting infection with this pathogen has been described. These tests may be classified as invasive and noninvasive or minimally invasive tests. Conventional tests such as histology and culture still play a major role in the diagnostic spectrum. Culture continues to be the only test allowing for a comprehensive analysis of pathogen characteristics and susceptibility to antibiotics. Urea breath test is the gold standard among noninvasive tests, with both stool antigen test and serology as appropriate substitutes or alternatives in specific situations. Among the multiple potential uses of molecular tests, noninvasive determination of resistance to antibiotics is of particular interest.
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Affiliation(s)
- Alexander M Hirschl
- Department of Clinical Microbiology, Institute of Hygiene and Medical Microbiology, Medical University, Vienna, Austria.
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Yamazaki T, Inoue M, Sasaki N, Yamada K, Minamide W, Matsumoto T. Clinical usefulness of culturing H. pylori from gastric juice in patients with severe neurological impairment. J Pediatr Gastroenterol Nutr 2004; 39:392-4. [PMID: 15448430 DOI: 10.1097/00005176-200410000-00016] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
OBJECTIVES To clarify the diagnostic usefulness of culturing Helicobacter pylori from gastric juice for the diagnosis of H. pylori infection in patients with neurologic impairment. METHODS Seventy-five patients (44 males, 31 females) with severe neurologic impairment admitted to Tokyo Children's Rehabilitation Hospital were included in this study. Samples of gastric juice obtained from these patients were put into Helicoporter, a transport medium for H. pylori. Samples were then centrifuged and sediment was inoculated onto a culture medium. Polymerase chain reaction assays were performed on 41 gastric juice samples for comparison with results of culture. Tests for occult blood in gastric juice and serum anti-H. pylori immunoglobulin G were tested for all patients. RESULTS H. pylori was isolated from 33 of 75 gastric juice samples by culture (positive rate, 44%). There was agreement between results of culture and polymerase chain reaction assays in 38 samples (93%). Twenty-two samples were positive and 16 were negative by both methods. In H. pylori immunoglobulin G-positive patients, occult blood was more frequently found in the culture-positive group (92%) than in the culture-negative group (59%) (P <0.025). The rate of detection of occult blood in immunoglobulin G-negative patients was similar in the culture-positive group (44%) and culture-negative group (50%). CONCLUSIONS The combination of a culture method using gastric juice and a serum H. pylori immunoglobulin G assay was not invasive and clinically useful for evaluating H. pylori infections among handicapped patients.
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Affiliation(s)
- Tsutomu Yamazaki
- Department of Pediatrics, Saitama Medical School, Saitama, Japan.
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Abstract
While there are some attempts to improve culture of Helicobacter pylori, molecular methods have been the main focus of this interest. Their main application concerns the development of rapid tests also allowing the determination of bacterial resistance, i.e. real-time polymerase chain reaction (PCR) or fluorescence in situ hybridization (FISH), or to genotype the strains. Attempts to improve, simplify or explain the discrepancies of urea breath test results have been made and new generation of stool antigen test with monoclonal antibodies either using the standard ELISA format or rapid immunoenzymatic detection have confirmed their value. With regard to serology, studies have mainly focused on the distinction of infections with more pathogenic strains and the ability to diagnose atrophic gastritis with the Gastropanel.
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Affiliation(s)
- Athanasios Makristathis
- Department of Clinical Microbiology, Institute of Hygiene and Medical Microbiology, Medical University Vienna, Vienna, Austria
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Shepherd AJ, Malcolm C, MacKay WG, Weaver LT. Childhood H. pylori: disappearing disease or chronic infection? Br J Community Nurs 2004; 9:201-5. [PMID: 15187900 DOI: 10.12968/bjcn.2004.9.5.12888] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 04/29/2023]
Abstract
Helicobacter pylori is one of the commonest chronic bacterial infections worldwide. It is acquired during childhood and its persistence has implications for health in later life. In adults, it is the principle cause of duodenal ulcer disease and there is evidence of an association between H. pylori and gastric cancer. However, most colonized people are asymptomatic. The prevalence of H. pylori increases with age but there is a striking difference between the rates in developed and developing countries. As no significant non-human or environmental source for this infection has been identified, person to person spread is almost certainly the main mode of transmission. Community nurses should be aware of this micro-organism as a potential cause of illness in children, and that they can play a role in promoting hygiene practices and educating families so that the risk of acquisition may be reduced. This review discusses the clinical features, prevalence, risk factors for transmission, diagnosis and treatment of H. pylori.
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Affiliation(s)
- A J Shepherd
- Department of Nursing and Midwifery, University of Stirling, Scotland, UK.
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