Comparison of real-time genotyping and quantitative PCR,multiplex-PCR and sequence analysis for hepatitis B virus genotypes B and C
10.3760/cma.j.issn.0254-5101.2010.12.020
- VernacularTitle:荧光定量分型PCR、多重PCR和测序检测HBV基因型的方法学比较
- Author:
Xiuyu ZHANG
;
Yao ZHAO
;
Wenlu ZHANG
;
Yuan HU
;
Zuowei YUAN
;
Ailong HUANG
- Publication Type:Journal Article
- Keywords:
Hepatitis B virus;
Genotype;
Real-time quantitative PCR;
Sequence analysis;
Multiplex-PCR
- From:
Chinese Journal of Microbiology and Immunology
2010;30(12):1154-1158
- CountryChina
- Language:Chinese
-
Abstract:
Objective To evaluate the real-time genotyping and quantitative PCR(RT-GQ-PCR)method by comparing it with direct sequence analysis and the multiplex-PCR method.Methods RT-GQ-PCR,direct sequence analysis and the multiplex-PCR method were used to detect HBV genotypes of 113 patient samples with HBV-DNA positive.ResultsThe detection rate of RT-GQ-PCR and direct sequence analysis was 100%,and the multiplex-PCR is 94.69%.The concordance between RT-GQ-PCR and the multiplex-PCR is perfect(Kappa value =0.915),and the consistency of RT-GQ-PCR and direct sequence analysis is pretty good(Kappa value = 0.742),specially at detecting single genotype.Twenty-eight samples with genotypes B and C dual infections were detected by RT-GQ-PCR,but only 19 samples by the multiplexPCR and 13 samples by direct sequence analysis.Conclusion The RT-GQ-PCR is convenient,rapid and accurate in HBV genotyping,especially more sensitive than direct sequence analysis and the multiplex-PCR for detecting dual genotypes.The method is applicable for large-scale epidemiological study.
