Department of Microbiology, Health Sciences Center, Kuwait University, Kuwait.
Virol J. 2010 May 30;7:111. doi: 10.1186/1743-422X-7-111.
Genotypes (A to H) of hepatitis B virus (HBV) influence liver disease progression and response to antiviral therapy in HBV-infected patients. Several methods have been developed for rapid genotyping of HBV strains. However, some of these methods may not be suitable for developing countries. The performance of INNO-LiPA HBV Genotyping assay (LiPA), direct DNA sequencing and subtractive PCR-RFLP of genotype-specific HBV genome regions were evaluated for accurately determining the HBV genotypes by analyzing sera (n = 80) samples from chronic HBV patients. Both, LiPA and DNA sequencing identified 63, 4 and 13 HBV strains as belonging to genotype D, genotype A and mixed genotype A and D, respectively. On the contrary, the PCR-RFLP-based method correctly identified all 4 genotype A but only 56 of 63 genotype D strains. Seven genotype D strains yielded indeterminate results. DNA sequence comparisons showed that a single nucleotide change in the target region generated an additional restriction site for Nla IV that compromised the accuracy of this method. Furthermore, all the mixed genotype A and D strains were identified only as genotype A strains. The data show that the PCR-RFLP-based method incorrectly identified some genotype D strains and failed to identify mixed genotype infections while LiPA and DNA sequencing yielded accurate results.
乙型肝炎病毒（HBV）的基因型（A 至 H）影响 HBV 感染患者的肝病进展和抗病毒治疗反应。已经开发了几种快速检测 HBV 株基因型的方法。然而，其中一些方法可能不适合发展中国家。通过分析来自慢性 HBV 患者的 80 份血清样本，评估 INNO-LiPA HBV 基因分型测定法（LiPA）、直接 DNA 测序和基因型特异性 HBV 基因组区域的减法 PCR-RFLP 在准确确定 HBV 基因型方面的性能。LiPA 和 DNA 测序均将 63、4 和 13 株 HBV 株分别鉴定为基因型 D、基因型 A 和基因型 A 和 D 的混合株。相反，基于 PCR-RFLP 的方法正确鉴定了所有 4 株基因型 A，但仅鉴定了 63 株基因型 D 中的 56 株。7 株基因型 D 株产生不确定结果。DNA 序列比较表明，目标区域的单个核苷酸变化产生了一个额外的 Nla IV 限制位点，从而影响了该方法的准确性。此外，所有的基因型 A 和 D 的混合株仅被鉴定为基因型 A 株。数据表明，基于 PCR-RFLP 的方法错误地鉴定了一些基因型 D 株，并且无法鉴定混合基因型感染，而 LiPA 和 DNA 测序则产生了准确的结果。
