[建立一种通过聚合酶链反应-限制性片段长度多态性（PCR-RFLP）对乙型肝炎病毒S区进行基因分型的新方法及其应用]

[Establishing a new genotyping method of hepatitis B virus by polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP) to analysis on S region and its application].

作者信息

Peng Liang, Ding Jing-Juan, Zhang Li-Sha

机构信息

Department of Infectious Diseases, Guiyang Medical College, Guiyang 550004, China.

出版信息

Zhonghua Gan Zang Bing Za Zhi. 2004 Aug;12(8):475-8.

PMID:15329207

Abstract

OBJECTIVE

To establish a new polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method of genotyping HBV using Mbo I, BsTN I, BsmA I, Hpa II and investigate the relationship between genotype and clinical spectrum of hepatitis B.

METHODS

124 full-genomic HBV sequences and 13 S-genomic sequences were analyzed, genotype specific regions were identified by the restriction enzymes Mbo I, BsTN I, BsmA I, Hpa II. And 176 samples from different kinds of hepatitis B were genotyped by this method. Five samples had been randomly selected and directly sequenced their S gene, to assess the accuracy.

RESULTS

In 176 serum samples of patients with hepatitis B from Guizhou area, genotype B and C were found in 56.8% and 43.2% respectively. The proportions of genotype B and C in ASC were 40.0% and 15.7% (chi-square = 12.16, P < 0.005); and they were 31.6% and 14.0% in CHB (chi-square = 7.88, P < 0.005).

CONCLUSION

Genotyping HBV, based on S gene RFLP seems to be highly sensitive, differential and accurate and could be used in large-scale surveys. HBV genotype B and C are existed in Guizhou area.

摘要

目的

建立一种利用Mbo I、BsTN I、BsmA I、Hpa II进行乙型肝炎病毒（HBV）基因分型的新型聚合酶链反应-限制性片段长度多态性（PCR-RFLP）方法，并研究HBV基因型与乙型肝炎临床谱之间的关系。

方法

分析124条HBV全基因组序列和13条S基因组序列，用限制性内切酶Mbo I、BsTN I、BsmA I、Hpa II鉴定基因型特异性区域。用该方法对176份不同类型乙型肝炎样本进行基因分型。随机选取5份样本对其S基因进行直接测序，以评估准确性。

结果

在贵州地区176例乙型肝炎患者血清样本中，B基因型和C基因型分别占56.8%和43.2%。B基因型和C基因型在慢性乙型肝炎表面抗原携带者（ASC）中的比例分别为40.0%和15.7%（卡方=12.16，P<0.005）；在慢性乙型肝炎（CHB）中分别为31.6%和14.0%（卡方=7.88，P<0.005）。