Title of article :
Quantification and genotyping of hepatitis B virus in a single reaction by real-time PCR and melting curve analysis
Author/Authors :
Shiou-Hwei Yeh، نويسنده , , Ching-Yi Tsai، نويسنده , , Jia-Horng Kao، نويسنده , , Chun-Jen Liu، نويسنده , , Ti-Jung Kuo، نويسنده , , Ming-Wei Lin، نويسنده , , Wen-Ling Huang، نويسنده , , Shu-Fen Lu، نويسنده , , Jane Jih، نويسنده , , Ding-Shinn Chen، نويسنده , , Pei-Jer Chen، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 2004
Pages :
8
From page :
659
To page :
666
Abstract :
Background/Aims Both viral titer and genotype of hepatitis B virus (HBV) play critical roles in determining clinical outcome and response to antiviral treatment in hepatitis B patients. In this study, a method was developed to determine both parameters in a single-tube reaction. Methods The method contains two consecutive steps, the first step used real-time PCR for quantification and second step used melting curve analysis for genotyping. For accurate quantification, the PCR primers and hybridization probes were selected from highly conserved regions to ensure the equivalent amplification and hybridization of all genotypes of HBVs. Within the sensor probe there exists signature single nucleotide polymorphisms (SNPs), which could effectively differentiate different HBV genotypes by showing different melting temperatures. Results The quantification results showed great consistency with the commercial assays in linear range from 102 to 1011 copies/ml. By comparison with the traditional restriction fragment length polymorphism (RFLP) methods, 99% of samples were accurately genotyped by current assay, and with a higher detection rate. In addition, this method can detect mixed HBV infections. Conclusions Currently, this methodology can be applied to areas prevalent with HBV genotypes B and C, providing an efficient alternative for clinical diagnosis and large-scaled longitudinal studies.
Keywords :
hepatitis B virus , Quantification , real-time PCR , genotyping
Journal title :
Journal of Hepatology
Serial Year :
2004
Journal title :
Journal of Hepatology
Record number :
586243
Link To Document :
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