Rapid detection of intestinal pathogens in fecal samples by an improved reverse dot blot method

doi:10.3748/wjg.15.2537

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May 28, 2009 (publication date) through Nov 24, 2024

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World Journal of Gastroenterology

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1007-9327

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Baishideng Publishing Group Inc, 7041 Koll Center Parkway, Suite 160, Pleasanton, CA 94566, USA

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World J Gastroenterol. May 28, 2009; 15(20): 2537-2542
Published online May 28, 2009. doi: 10.3748/wjg.15.2537

Rapid detection of intestinal pathogens in fecal samples by an improved reverse dot blot method

Jian-Ming Xing, Su Zhang, Ying Du, Dan Bi, Li-Hui Yao

Jian-Ming Xing, Su Zhang, Ying Du, Dan Bi, Li-Hui Yao, Huzhou Maternity and Child Care Hospital, Huzhou 313000, Zhejiang Province, China

Author contributions: Xing JM and Zhang S carried out all the experiments and drafted the manuscript; Du Y designed oligonucleotide microarray procedure described here; Bi D collected and identified the clinical specimens by using a conventional assay; Yao LH performed the statistical analysis; All authors have read and approved the final manuscript.

Correspondence to: Jian-Ming Xing, Huzhou Maternity and Child Care Hospital, Huzhou 313000, Zhejiang Province, China. xjm2161360@163.com

Telephone: +86-572-2030381

Fax: +86-572-2030109

Received: February 15, 2009
Revised: April 12, 2009
Accepted: April 19, 2009
Published online: May 28, 2009

Abstract

AIM: To develop a new, rapid and accurate reverse dot blot (RDB) method for the detection of intestinal pathogens in fecal samples.

METHODS: The 12 intestinal pathogens tested were Salmonella spp., Brucella spp., Escherichia coli O157:H7, Clostridium botulinum, Bacillus cereus, Clostridium perfringens, Vibrio parahaemolyticus, Shigella spp., Yersinia enterocolitica, Vibrio cholerae, Listeria monocytogenes and Staphylococcus aureus. The two universal primers were designed to amplify two variable regions of bacterial 16S and 23S rDNA genes from all of the 12 bacterial species tested. Five hundred and forty fecal samples from the diarrhea patients were detected using the improved RDB assay.

RESULTS: The methods could identify the 12 intestinal pathogens specifically, and the detection limit was as low as 103 CFUs. The consistent detection rate of the improved RDB assay compared with the traditional culture method was up to 88.75%.

CONCLUSION: The hybridization results indicated that the improved RDB assay developed was a reliable method for the detection of intestinal pathogen in fecal samples.

Keywords: Immunoblotting; Intestinal pathogens; Feces