Yang Jie, Dai Lin, Guo Ya-Bing, Yang Shou-Chang, Wang Yan-Jun, Luo Kang-Xian
Department of Infectious Diseases, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China.
Di Yi Jun Yi Da Xue Xue Bao. 2002 Aug;22(8):707-9.
To establish a convenient method for the genotyping of hepatitis B virus (HBV) using multiplex PCR.
Based on the alignment of 114 complete nucleotide sequences of HBV DNA belonging to different genotypes, acquired from the GenBank, genotype-specific sequences were identified according to which 6 pairs of primers were designed corresponding to each genotype. Subsequent genotyping of HBV was performed using these primers that were added, either alone or in conjunction with others, into a multiplex PCR reaction tube, and HBV genotype was determined according to the length of amplified DNA.
The genotyping result of multiplex PCR was consistent with that produced by PCR- restriction fragment length polymorphism as established by Lindh. We found in this study that among the HBV carriers in the vicinities Guangzhou of City, about 45% belonged to B genotype, 38.75% to C genotype and 16.75% to D genotype.
This multiplex PCR method is simple, convenient and more differential.
建立一种利用多重聚合酶链反应（PCR）对乙型肝炎病毒（HBV）进行基因分型的简便方法。
基于从基因库获取的114条不同基因型HBV DNA的完整核苷酸序列比对，鉴定出基因型特异性序列，并据此设计了6对分别对应各基因型的引物。随后，将这些引物单独或与其他引物组合加入多重PCR反应管中进行HBV基因分型，并根据扩增DNA的长度确定HBV基因型。
多重PCR的基因分型结果与Lindh建立的PCR-限制性片段长度多态性方法的结果一致。我们在本研究中发现，广州市周边地区的HBV携带者中，约45%属于B基因型，38.75%属于C基因型，16.75%属于D基因型。
这种多重PCR方法简单、方便且鉴别力更强。
