Simultaneous Genotyping and Quantification of Hepatitis B Virus for Genotypes B and C by Real-Time PCR Assay
Simultaneous Genotyping and Quantification of Hepatitis B Virus for Genotypes B and C by Real-Time PCR Assay
复制标题
通过实时 PCR 检测同时对 B 型和 C 型乙型肝炎病毒进行基因分型和定量
DOI：
10.1128/jcm.00741-10
复制
发表时间：
2010-10-01
影响因子：
9.4
通讯作者：
Huang, Ai-Long
中科院分区：
文献类型：
--
作者：
Zhao, Yao;Zhang, Xiu-Yu;Huang, Ai-Long
B型肝炎病毒（HBV）是人类慢性肝病的重要病因，是一个重大的公共卫生问题。病毒载量和HBV基因型在决定B型肝炎患者的临床结局和抗病毒治疗反应方面起着关键作用。病毒基因型检测和定量测定目前正在使用，具有不同的有效性水平。在本研究中，评价了基于实时基因分型和定量PCR（GQ-PCR）的检测方法的性能。通过使用基因型特异性引物和探针，该检测试剂盒可在单一反应中同时鉴定和定量基因型B和C。我们的GQ-PCR正确鉴定了所有预定义的基因型B和C，并且没有观察到基因型B和C之间的交叉反应。GQ-PCR比直接测序法检出更多的B和C混合基因型HBV感染者。用GQ-PCR方法对127例HBV感染者进行基因分型，发现56.7%的HBV为B型，13.4%为C型，29.8%为B和C混合型。该检测方法提供了一种可靠、有效和经济有效的方法，用于定量单一或混合感染中HBV的B和C基因型。该检测适用于连续监测病毒载量水平，并确定亚洲人基因型病毒载量与疾病分期之间的关系。
Hepatitis B virus (HBV) is an important cause of human chronic liver diseases and is a major public health problem. Viral load and HBV genotype play critical roles in determining clinical outcomes and response to antiviral treatment in hepatitis B patients. Viral genotype detection and quantification assays are currently in use with different levels of effectiveness. In this study, the performance of a real-time genotyping and quantitative PCR (GQ-PCR)-based assay was evaluated. Through the use of genotype-specific primers and probes, this assay provides simultaneous identification and quantification of genotypes B and C in a single reaction. Our GQ-PCR correctly identified all predefined genotypes B and C, and no cross-reaction between genotypes B and C were observed. The GQ-PCR identified more cases of HBV infections with mixed genotypes B and C than direct sequencing did. Samples from 127 HBV-infected Chinese patients were genotyped with GQ-PCR, revealing 56.7% HBV as genotype B, 13.4% as genotype C, and 29.8% as mixed genotypes B and C. This assay provides a reliable, efficient, and cost-effective means for quantification of the B and C genotypes of HBV in single or mixed infections. This assay is suitable for sequential monitoring of viral load levels and for determining the relationship between the genotype viral load and stage of disease in Asians.