Huang Hai-yan, Meng Xiang-wei, Zhang Ling-ling
No. 1 Hospital of Jilin University, Changchun 130021, China.
Zhonghua Liu Xing Bing Xue Za Zhi. 2006 Dec;27(12):1057-60.
To study the hepatitis B virus (HBV) genotype and its relation to clinical degree and responsiveness to antiviral therapy on hepatitis in order to guide the clinical therapy.
We amplified HBV S gene by polymerase chain reaction (PCR), using the second-round PCR product, which was digested by restriction fragment length polymorphism (RFLP). This genotype method was designed under the analysis of the restriction fragment length polymorphism and using the restriction enzymes that identified the genotype-specific sequences. Five restriction enzymes, Hph I , Nci I , Alw I, Ear I and NlaIV, were identified in genotype-specific RFLP from the S gene region. Representative sequences from the S genome region of each HBV genotype were aligned to show the restriction sites by the five restriction enzymes. The amplified S gene nucleotide sequences were sequenced by dideoxy-chain-termination method and the corresponding amino acid sequence was deduced using DNASIS software. Later, they were genotyped by comparing to representative S gene sequences obtained from GenBank. This confirmed the results of RFLP HBV genotyping methods, coincident with that of S gene sequence.
Genotypes A, B, C, D were classified in 216 patients with HBV and DNA positive. The results showed that: 1 case (0.46%) of genotype A, 19 cases genotype B (8.8% ), 175 genotype C (81.02%) and 21 genotype D (9.72%). A total of 86 patients in the hospital were divided into either genotype C cases (69) or non-genotype C cases (17).
Genotype C was the major genotype in Changchun. Among HBV patients, type C was 80.95%, followed by genotypes B and D. Both hepatocellular carcinoma and liver cirrhosis showed relations with genotype C.
研究乙型肝炎病毒（HBV）基因型及其与肝炎临床程度和抗病毒治疗反应性的关系，以指导临床治疗。
我们通过聚合酶链反应（PCR）扩增HBV S基因，使用第二轮PCR产物，通过限制性片段长度多态性（RFLP）进行消化。这种基因型方法是在分析限制性片段长度多态性并使用识别基因型特异性序列的限制性内切酶的基础上设计的。从S基因区域的基因型特异性RFLP中鉴定出五种限制性内切酶，即Hph I、Nci I、Alw I、Ear I和NlaIV。对每种HBV基因型S基因组区域的代表性序列进行比对，以显示这五种限制性内切酶的限制性位点。通过双脱氧链终止法对扩增的S基因核苷酸序列进行测序，并使用DNASIS软件推导相应的氨基酸序列。随后，通过与从GenBank获得的代表性S基因序列进行比较对它们进行基因分型。这证实了RFLP HBV基因分型方法的结果，与S基因序列的结果一致。
在216例HBV DNA阳性患者中鉴定出A、B、C、D基因型。结果显示：A基因型1例（0.46%），B基因型19例（8.8%），C基因型175例（81.02%），D基因型21例（9.72%）。医院共86例患者分为C基因型组（69例）和非C基因型组（17例）。
C基因型是长春地区的主要基因型。在HBV患者中，C型占80.95%，其次是B型和D型。肝细胞癌和肝硬化均与C基因型有关。
