Rapid Communication
Copyright ©2008 The WJG Press and Baishideng. All rights reserved.
World J Gastroenterol. Jan 7, 2008; 14(1): 120-124
Published online Jan 7, 2008. doi: 10.3748/wjg.14.120
Comparison of ligase detection reaction and real-time PCR for detection of low abundant YMDD mutants in patients with chronic hepatitis B
Xiao-Ling Wang, Song-Gang Xie, Ling Zhang, Wei-Xia Yang, Xing Wang, Hong-Zhi Jin
Xiao-Ling Wang, Song-Gang Xie, Ling Zhang, Wei-Xia Yang, Laboratory Center, Subei People’s Hospital, Yangzhou 225001, Jiangsu Province, China
Xing Wang, Department of Infectious Diseases, Subei People’s Hospital, Yangzhou 225001, Jiangsu Province, China
Hong-Zhi Jin, School of Life Science, Shanghai Jiaotong University, Shanghai 200240, China
Correspondence to: Xiao-Ling Wang, Laboratory Center, Subei People’s Hospital,98 West Nantong Road, Yangzhou 225001, Jiangsu Province, China. wxlf26@163.com
Telephone: +86-514-7115256
Fax: +86-514-7937455
Received: August 1, 2007
Revised: October 13, 2007
Published online: January 7, 2008
Abstract

AIM: To compare the ligase detection reaction (LDR) and real-time PCR for detection of low abundant YMDD mutants in patients with chronic hepatitis B infection.

METHODS: Mixtures of plasmids and serum samples from 52 chronic hepatitis B patients with low abundant lamivudine-resistant mutations were tested with LDR and real-time PCR. Time required and reagent cost for both assays were evaluated.

RESULTS: Real-time PCR detected 100, 50, 10, 1 and 0.1% of YIDD plasmid, whereas LDR detected 100, 50, 10, 1, 0.1, and 0.01% of YIDD plasmid, in mixtures with YMDD plasmid of 106 copies/mL. Among the 52 clinical serum samples, completely concordant results were obtained for all samples by both assays, and 39 YIDD, 9 YVDD, and 4 YIDD/YVDD were detected. Cost and time required for LDR and real-time PCR are 60/80 CNY (8/10.7 US dollars) and 4.5/2.5 h, respectively.

CONCLUSION: LDR and real-time PCR are both sensitive and inexpensive methods for monitoring low abundant YMDD mutants during lamivudine therapy in patients with chronic hepatitis B. LDR is more sensitive and less expensive, while real-time PCR is more rapid.

Keywords: YMDD mutants; Hepatitis B virus; Real-time PCR; Ligase detection reaction