Hussain Munira, Chu Chi-Jen, Sablon Erwin, Lok Anna S F
Division of Gastroenterology, University of Michigan Medical Center, Ann Arbor, Michigan 48109-0362, USA.
J Clin Microbiol. 2003 Aug;41(8):3699-705. doi: 10.1128/JCM.41.8.3699-3705.2003.
Hepatitis B virus (HBV) genotypes may influence HBeAg seroconversion rates, mutational patterns in the precore (PC) and core promoter (CP) regions, severity of liver disease, and response to antiviral treatment. Development of rapid, simple, and standardized assays to detect viral genotypes and common mutations in the PC and CP regions can accelerate research on the clinical significance of these variants. We aim to assess the accuracy of a line probe assay in determining HBV genotypes and detecting HBV PC and CP variants. HBV genotypes in 701 patients and PC and CP variants in 600 patients with chronic HBV infection from China and the United States were studied using the INNO-LiPA assay. All but one (99.9%) sample were classified by the genotyping assay. All eight genotypes, i.e., A to H, were found. The INNO-LiPA genotyping assay results were completely concordant with those of sequencing. Using the INNO-LiPA PC assay, 99.8 and 94.7% samples were classifiable in the PC and CP regions, respectively. The PC assay results were completely concordant with those of sequencing in all samples that showed either wild-type or variant sequence. The line probe assay was more sensitive in detecting mixtures than was direct sequencing. By INNO-LiPA, only 50 and 27% of the samples, with mixed wild-type and variant sequence in the PC and CP region, respectively, showed mixed sequence by direct sequencing. INNO-LiPA is rapid, sensitive, and reliable-thus enabling accurate determination of HBV genotypes and detection of PC and CP variants in a large population of patients.
乙型肝炎病毒（HBV）基因型可能会影响HBeAg血清学转换率、前核心（PC）区和核心启动子（CP）区的突变模式、肝脏疾病的严重程度以及对抗病毒治疗的反应。开发快速、简便且标准化的检测方法来检测病毒基因型以及PC区和CP区的常见突变，能够加速对这些变异体临床意义的研究。我们旨在评估一种线性探针检测法在确定HBV基因型以及检测HBV PC和CP变异体方面的准确性。使用INNO-LiPA检测法对来自中国和美国的701例患者的HBV基因型以及600例慢性HBV感染患者的PC和CP变异体进行了研究。除1个样本外（99.9%），所有样本均通过基因分型检测进行了分类。发现了所有8种基因型，即A至H型。INNO-LiPA基因分型检测结果与测序结果完全一致。使用INNO-LiPA PC检测法，分别有99.8%和94.7%的样本在PC区和CP区可进行分类。在所有显示野生型或变异序列的样本中，PC检测结果与测序结果完全一致。线性探针检测法在检测混合序列方面比直接测序更敏感。通过INNO-LiPA检测，在PC区和CP区野生型和变异序列混合的样本中，分别只有50%和27%的样本通过直接测序显示出混合序列。INNO-LiPA检测法快速、灵敏且可靠，因此能够在大量患者中准确确定HBV基因型并检测PC和CP变异体。
