Brief Reports
Copyright ©The Author(s) 2004. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Aug 15, 2004; 10(16): 2439-2443
Published online Aug 15, 2004. doi: 10.3748/wjg.v10.i16.2439
Simultaneous detection of HBV and HCV by multiplex PCR normalization
Ning Wang, Xue-Qin Gao, Jin-Xiang Han
Ning Wang, Xue-Qin Gao, Jin-Xiang Han, Shandong Medicinal Biotechnological Center, Shandong Academy of Medical Sciences; Key Laboratory for Biotechdrugs, Ministry of Public Health, Jinan 250062, Shandong Province, China
Author contributions: All authors contributed equally to the work.
Supported by the National Key Technology Research and Development Program of China during the 9th Five-Year Plan Period, No. 96C020117
Correspondence to: Professor Jin-Xiang Han, Shandong Medicinal Biotechnological Center, Shandong Academy of Medical Sciences; Key Laboratory for Biotechdrugs, Ministry of Public Health, Jinan 250062, Shandong Province, China. jxhan@sdu.edu.cn
Telephone: +86-531-2919888
Received: August 23, 2003
Revised: September 18, 2003
Accepted: October 7, 2003
Published online: August 15, 2004
Abstract

AIM: To design and establish a method of multiplex PCR normalization for simultaneously detecting HBV and HCV.

METHODS: Two pairs of primers with a 20 bp joint sequence were used to amplify the target genes of HBV and HCV by two rounds of amplification. After the two rounds of amplification all the products had the joint sequence. Then the joint sequence was used as primers to finish the last amplification. Finally multiplex PCR was normalized to a single PCR system to eliminate multiplex factor interference. Four kinds of nucleic acid extraction methods were compared and screened. A multiplex PCR normalization method was established and optimized by orthogonal design of 6 key factors. Then twenty serum samples were detected to evaluate the validity and authenticity of this method.

RESULTS: The sensitivity, specificity, diagnostic index and efficiency were 83.3%, 70%, 153.3% and 72.2%, respectively for both HBsAg and anti-HCV positive patients, and were 78.6%, 80%, 258.6% and 79.2%, respectively for HBsAg positive patients, and were 75%, 90%, 165% and 83.3%, respectively for anti-HCV positive patients.

CONCLUSION: The multiplex PCR normalization method shows a broad prospect in simultaneous amplification of multiple genes of different sources. It is practical, correct and authentic, and can be used to prevent and control HBV and HCV.

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