Classification of hepatitis B virus genotypes by the PCR-Invader method with genotype-specific probes

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• 作者：Kenichi Tadokoro ; Mariko Kobayashi ; Toshikazu Yamaguchi ; Fumitaka Suzuki ; Saeko Miyauchi ; Toru Egashira and Hiromitsu Kumada
• 关键词：Hepatitis B virus ; PCR-Invader ; Subgenotype ; Ba ; Bj
• 刊名：Journal of Virological Methods
• 出版年：2006
• 出版时间：December 2006
• 年：2006
• 卷：138
• 期：1-2
• 页码：30-39
• 全文大小：1558 K

文摘

Hepatitis B virus is a worldwide public health problem. A simple and effective test to identify viral genotypes would greatly aid efforts to understand and control the spread of this disease. A serial invasive signal amplification reaction assay (PCR-Invader assay) was developed for distinguishing the known eight genotypes (A–H) and four subgenotypes (Aa, Ae, Ba, Bj) of hepatitis B virus (HBV). The preS/S and core regions were amplified by multiplex PCR and delivered to 12 wells containing genotype-specific Invader probes. By observing the fluorescence patterns in the wells, HBV sub/genotypes can be assigned. A total of 505 serum samples containing HBV/HBsAg in Japan was examined by PCR-Invader and compared the results with those from ELISA assays with monoclonal antibodies against epitopes on gene products of the preS2 region and with a genotype-specific probe assay (GSPA) based on the preS1 region. Genotypes determined by the PCR-Invader agreed with those of the ELISA method in 98.2 % of cases and with the GSPA method in 97.1 % of cases. Co-infection with two distinct genotypes was correctly identified by the PCR-Invader in four serum samples, as determined by GSPA. Thus, the PCR-Invader assay is a useful tool for detecting the 10 known HBV sub/genotypes.