Qian WP, Tan YQ, Chen Y, Peng Y, Li Z, Lu GX, Lin MC, Kung HF, He ML, Shing LK. Rapid quantification of semen hepatitis B virus DNA by real-time polymerase chain reaction. World J Gastroenterol 2005; 11(34): 5385-5389 [PMID: 16149152 DOI: 10.3748/wjg.v11.i34.5385]
Corresponding Author of This Article
Dr. Ming-Liang He, The Center for Emerging Infectious Diseases, The Chinese University of Hong Kong, Hong Kong Special Administrative Region, China. mlhe@cuhk.edu.hk
Article-Type of This Article
Brief Reports
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This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Wei-Ping Qian, Luofu Hospital, Shenzhen 518033, Guangdong Province, China
Wei-Ping Qian, Yue-Qiu Tan, Guang-Xiu Lu, Institute of Reproduction and Stem Cell Engineering, Xiangya School of Medicine, Central South University, Changsha 410008, Hunan Province, China
Zhi Li, Marie C. Lin, The Institute of Molecular Biology, The University of Hong Kong, Hong Kong Special Administrative Region, China
Ying Peng, Department of Neurology, The Second Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510120, Guangdong Province, China
Ying Chen, Hsiang-Fu Kung, Ming-Ling He, Li-Ka Shing, The Center for Emerging Infectious Diseases, The Chinese University of Hong Kong, Hong Kong Special Administrative Region, China
Author contributions: All authors contributed equally to the work.
Supported by Research Fund for the Control of Infectious Diseases and Research Grant Committee of Hong Kong Government
Correspondence to: Dr. Ming-Liang He, The Center for Emerging Infectious Diseases, The Chinese University of Hong Kong, Hong Kong Special Administrative Region, China. mlhe@cuhk.edu.hk
Telephone: +852-2252-8813 Fax: +852-2635-4977
Received: September 14, 2004 Revised: January 23, 2005 Accepted: January 26, 2005 Published online: September 14, 2005
Abstract
AIM: To examine the sensitivity and accuracy of real-time polymerase chain reaction (PCR) for the quantification of hepatitis B virus (HBV) DNA in semen.
METHODS: Hepatitis B viral DNA was isolated from HBV carriers’ semen and sera using phenol extraction method and QIAamp DNA blood mini kit (Qiagen, Germany). HBV DNA was detected by conventional PCR and quantified by TaqMan technology-based real-time PCR (quantitative polymerase chain reaction (qPCR)). The detection threshold was 200 copies of HBV DNA for conventional PCR and 10 copies of HBV DNA for real time PCR per reaction.
RESULTS: Both methods of phenol extraction and QIAamp DNA blood mini kit were suitable for isolating HBV DNA from semen. The value of the detection thresholds was 500 copies of HBV DNA per mL in the semen. The viral loads were 7.5 × 107 and 1.67 × 107 copies of HBV DNA per mL in two HBV infected patients’ sera, while 2.14 × 105 and 3.02 × 105 copies of HBV DNA per mL in the semen.
CONCLUSION: Real-time PCR is a more sensitive and accurate method to detect and quantify HBV DNA in the semen.
