摘要
目的:采用多对型特异性引物,通过巢式PCR法检测湖南省乙肝患者血清中乙型肝炎病毒(HBV)基因型的分布情况.方法:根据从前S1基因到S基因中的保守序列设计出10条内外引物,并将其中8条内引物分成A、B两组,分别扩增A、B、C和D、E、F型HBV,然后将第2轮PCR的两组产物分别用3%琼脂糖进行电泳,根据PCR产物片断大小直接判定HBV基因型.与目前常用的PCR-RFLP法进行了比较,并做重复试验以证实该方法的可靠性和准确性.用此法检测了220例湖南籍慢性乙肝血清中的HBV基因型,以了解湖南人群的HBV基因型分布情况.结果:多对型特异性引物巢式PCR与PCR-RFLP法的检测结果完全一致,重现率(100%);湖南人群HBV基因分型结果为B型190例(86.4%)、C型30例(13.6%).结论:这种新的巢式PCR分型法能清晰直观地辨别HBV基因型,结果准确可靠.用此法证实了湖南人群的HBV优势基因型以B型为主,C型次之.
AIM: To determine the genotypes of hepatitis B virus (HBV) in Hunan Province of China by nested PCR with multiplex pairs of genotype-specific primers. METHODS: Ten outer and inner primers were designed on the basis of the conserved nature of nuleotide sequences in regions of the Pre-S1 through S genes, in which 8 inner primers were devided into mix A and B to amplify HBV of genotype A, B, C and D, E, F respectively. The two different products from one sample in second-round PCR were separately electrophoresed on a 3% agarose gel. Genotypes of HBV were determined directly by the size of PCR products. To test its reliability and veracity, we compared new nested PCR with popular PCR-RFLP, followed by repeated experiments. This nested PCR was also used in the genotyping of HBVs in 220 Hunan patients with chronic hepatitis B to know the distribution of HBV genotype in Hunan Province of China. RESULTS: The results showed complete concordence between the two assays and 100% recurrence in the repeated experiments. Of the 220 Hunan patients, 190 (86.4%) were genotype B and 30 (13.6%) were genotype C. CONCLUSION: This new nested PCR can help to determine HBV genotypes clearly and directly with reliable and accurate results. With the application of this new method, the predominant HBV genotypes in Hunan are confirmed to be genotypes B and C.
出处
《世界华人消化杂志》
CAS
2004年第2期332-335,共4页
World Chinese Journal of Digestology
