中国应用抗 B、C 基因型抗体的 ELISA 基因分型检测乙型肝炎病毒。

ELISA genotyping of hepatitis B virus in China with antibodies specific for genotypes B and C.

机构信息

Key Laboratory of Bionic Engineering (Ministry of Education), Jilin University, Changchun, 130022, China.

Beijing Abace Biotechnology, Beijing, 100176, China.

出版信息

Sci Rep. 2024 Oct 12;14(1):23884. doi: 10.1038/s41598-024-76023-7.

DOI:10.1038/s41598-024-76023-7

PMID:39396069

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11470951/

Abstract

Hepatitis B virus (HBV) causes hepatitis B (HB) and distinct HBV genotypes can lead to different prognoses. However, HBV genotyping is rarely done in clinics, because the traditional method by PCR-based DNA sequencing is impractical for clinical diagnosis with tedious process and low success rate. Herein, we have established an ELISA-based genotyping method to quickly determine the HBV genotypes of HB patients in China. First, two commercial antibodies, 16D12 and 6H3 specific for HBV genotypes B and C respectively, are chosen as capture antibodies, since these two genotypes dominate in China. Then two home-made genotype-specific antibodies, B19 and C04, are used as the detection antibodies for genotypes B and C in sandwiched ELISA. The ELISA kit shows high sensitivity (> 95%) and specificity (> 95%) in detecting genotypes B and C of Chinese HB patients. Moreover, the ELISA kit has demonstrated higher success rate (98.7%) than PCR-based DNA sequencing (93.5%) and a commercial PCR-based genotyping kit (92.2%) for sera with HBV DNA ≥ 1000 IU/mL and HBsAg ≥ 250 IU/mL. Such an advantage is more obvious for the sera with HBV DNA < 1000 IU/mL. The kappa analysis between the ELISA and PCR-based DNA sequencing results exhibits a kappa of 0.836, indicating a good correlation.

摘要

乙型肝炎病毒（HBV）可引起乙型肝炎（HB），不同的 HBV 基因型可导致不同的预后。然而，HBV 基因型分析在临床上很少进行，因为基于 PCR 的 DNA 测序的传统方法在临床诊断中操作繁琐、成功率低，不切实际。在此，我们建立了一种基于 ELISA 的基因型分析方法，用于快速确定中国 HB 患者的 HBV 基因型。首先，选择两种针对基因型 B 和 C 的商用抗体 16D12 和 6H3 作为捕获抗体，因为这两种基因型在中国占主导地位。然后，使用两种自制的基因型特异性抗体 B19 和 C04 作为夹心 ELISA 检测基因型 B 和 C 的检测抗体。该 ELISA 试剂盒在检测中国 HB 患者的基因型 B 和 C 时具有较高的灵敏度（>95%）和特异性（>95%）。此外，与基于 PCR 的 DNA 测序（93.5%）和商业的基于 PCR 的基因分型试剂盒（92.2%）相比，该 ELISA 试剂盒在 HBV DNA≥1000 IU/mL 和 HBsAg≥250 IU/mL 的血清中具有更高的成功率（98.7%）。对于 HBV DNA<1000 IU/mL 的血清，这种优势更加明显。ELISA 与基于 PCR 的 DNA 测序结果之间的 Kappa 分析显示 Kappa 值为 0.836，表明相关性良好。