反向斑点杂交技术检测HBV基因型方法的评价

Evaluation of reverse dot blot assay for Hepatitis B virus genotyping

目的评价用于检测乙型肝炎病毒(HBV)基因型的反向斑点杂交(RDB)方法。方法应用RDB检测723例HBV阳性标本的HBV基因型。其中对423例HBV序列测序结果清晰可读的样本进一步使用NCBI在线数据库分析、系统进化法分析其HBV基因型,并与RDB法结果进行比较。将样本HBV序列与所用的探针序列进行比对,以评价单个碱基突变对该方法的影响。结果 RDB法检测的检出率为97.6%。RDB检测结果与NCBI在线数据库分析法的一致性为98.8%;与系统进化分析法一致性为100%。与对应探针有1个碱基差异的HBV A型1例、B型12例、C型46例和D型1例均被成功检出。结论 RDB是一种灵敏、准确、有效,且具有很高临床应用价值的HBV基因型检测方法。单碱基的突变不会影响其对HBV的分型结果。

Objective To evaluate the reverse dot blot（RDB） assay for Hepatitis B virus（HBV） genotyping.Methods The HBV genotyping in 723 HBV samples was determined by RDB assay.423 samples of clear sequencing result were further analyzed by National Center for Biotechnology Information（NCBI） online database,and the HBV genotyping was analyzed systematically.The results were compared with those by RDB.The influence of single-nucleotide mutation on RDB was evaluated,and the HBV sequences were compared with probe sequences.Results The positive detection rate was 97.6%.The concordance of RDB and NCBI online sequencing method was 98.8%,and the concordance of RDB and systematical analysis method was 100%.1 Genotype A,12 Genotype B,46 Genotype C and 1 Genotype D were found 1 nucleotide mismatch to corresponding probe.Conclusions The RDB method is sensitive,accurate and effective for clinical routine HBV genotyping.It is insensitive to single-nucleotide mutation.