乙型肝炎病毒C/D重组体毒株的三种鉴定方法

目的针对HBV C/D重组体在我国西部地区尤其是藏族人群中广泛流行,基因分型时易被错分为D基因型等问题,建立相对准确的鉴定C/D重组体的方法。方法以来自西部地区的25例HBV感染者为例,分别通过PCR片段长度法、S和前C/C基因序列进化树分析法、全基因序列精细分析法对C/D重组体进行鉴定比较。结果 D基因型存在33 bp缺失,在S基因区酶切或测序的基础上,通过跨缺失区段的PCR产物长度可区分D与C/D重组体,该法简单易操作,适于大样本初筛;S和前C/C基因序列进化树分析法可明确鉴定C/D重组体,准确度高;全基因序列精细分析法可对重组类型和重组位点进行精确分析,结果最为可靠。结论根据研究目的不同,应综合考虑运用几种C/D重组体鉴定方法,提高研究的效率和结果的可靠性。

Three methods to identify hepatitis B virus strains with C/D recombinant

Objective To investigate accurate method for identifying hepatitis B virus strains with C/D recombinant.Methods A total of 25 patients with chronic hepatitis B from western China were enrolled in this study.Three methods, containing polymerase chain reaction (PCR) of fragment length analysis, phylogenetic analysis based on S and preC/C gene, and full genome precise analysis, were compared.Results On account of the 33bp deletion in preS1 of genotype D, a PCR fragment spanning the preS1 region could identify C/D recombinant from genotype D, which was easy to perform and suitable for large sample screening.Phylogenetic analysis based on S and preC/C gene could identify C/D recombinant with high accuracy.Full genome precise analysis could identify C/D recombinant precisely and accurately which was the most reliable method.Conclusion For various research purposes, the application of the 3 methods should be performed comprehensively to improve the efficiency and reliability in identification of C/D recombinant.