Abstract

For personalized treatment of chronic HBV-infected patients, it is necessary to identify hepatitis B virus (HBV) surface antigen (HBsAg) subtypes and HBV genotypes circulating among infected individuals. To this end, a laboratory version of the test kit for HBsAg subtype and HBV genotype identification in human blood serum/plasma using enzyme-linked immunosorbent assay (ELISA) and a custom monoclonal antibody (MAb) panel has been developed. With this kit version, it appears possible to identify the main HBsAg subtypes (ayw2, ayw3, adw2, and adrq+) and HBV genotypes (A, C, and D) circulation in Russian Federation. The aim of the work was to test the laboratory-developed ELISA kits for HBsAg subtyping and HBV genotyping in human HBsAg-positive blood plasma samples. A total of 146 HBsAg-positive plasma samples were tested. For 108 samples with detectable viral DNA, the results of HBsAg subtyping and HBV genotyping using the suggested MAb panel were compared with the results of genomic DNA sequencing for HBV isolates. For 38 HBsAg-positive samples in which HBV DNA could not be detected, ELISA, using the highly specific MAb panel kindly provided by P. Swenson, PhD, was employed as a reference technique. The study demonstrated that for the group of HBsAg-positive samples with detectable HBV DNA, the coincidence between the results obtained using the suggested MAb panel and the results of HBV DNA genome sequence analysis was 85% for HBsAg subtyping and 98% for HBV genotyping. For the group of HBsAg-positive samples with undetectable HBV DNA, the coincidence between the two techniques (the one using the suggested MAb panel and one using Swenson’s MAb panel) was 95% in the case of HBsAg subtyping and 100% for HBV genotyping. The reported results allow recommending the suggested laboratory ELISA test kit for HBsAg subtype and HBV genotype identification in human blood plasma as an alternative or a complementary technique to genomic analysis in order to study HBV characteristics especially in those cases when HBV DNA isolation appears impossible.

中文翻译：

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人体血浆中HBsAg亚型和乙型肝炎病毒基因型鉴定的ELISA检测试剂盒的试验结果

摘要

为了对慢性HBV感染的患者进行个性化治疗，有必要确定在感染个体之间传播的乙型肝炎病毒（HBV）表面抗原（HBsAg）亚型和HBV基因型。为此，已开发出实验室版本的试剂盒，用于使用酶联免疫吸附测定（ELISA）和定制单克隆抗体（MAb）面板鉴定人血清/血浆中的HBsAg亚型和HBV基因型。使用此套件版本，似乎可以识别俄罗斯联邦的主要HBsAg亚型（ayw2，ayw3，adw2和adrq +）和HBV基因型（A，C和D）。这项工作的目的是测试实验室开发的用于检测人HBsAg阳性血浆样本中HBsAg亚型和HBV基因型的ELISA试剂盒。总共测试了146个HBsAg阳性血浆样品。对于108个具有可检测的病毒DNA的样品，将使用建议的MAb面板进行HBsAg亚型分型和HBV基因分型的结果与HBV分离株的基因组DNA测序的结果进行了比较。对于38个无法检测到HBV DNA的HBsAg阳性样品，采用P.Swenson博士提供的高度特异性MAb试剂盒的ELISA作为参考技术。该研究表明，对于具有可检测的HBV DNA的HBsAg阳性样品组，使用建议的MAb面板获得的结果与HBV DNA基因组序列分析的结果之间的一致性为HBsAg亚型为85％，HBV基因型为98％。对于检测不到HBV DNA的HBsAg阳性样品，在HBsAg亚型分型的情况下，两种技术（使用建议的MAb组的一种技术和使用Swenson's MAb组的一种技术）的一致性为95％，对于HBV基因分型为100％。报告的结果允许推荐建议的实验室ELISA测试试剂盒，用于鉴定人血浆中的HBsAg亚型和HBV基因型，作为基因组分析的替代或补充技术，以便研究HBV特性，尤其是在似乎无法分离HBV DNA的情况下。