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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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f93/11470951/8992380d401d/41598_2024_76023_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f93/11470951/01934acc7562/41598_2024_76023_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f93/11470951/d845abb7ef72/41598_2024_76023_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f93/11470951/21de9d93e6fd/41598_2024_76023_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f93/11470951/59fc7fdac6b4/41598_2024_76023_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f93/11470951/8992380d401d/41598_2024_76023_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f93/11470951/01934acc7562/41598_2024_76023_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f93/11470951/d845abb7ef72/41598_2024_76023_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f93/11470951/21de9d93e6fd/41598_2024_76023_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f93/11470951/59fc7fdac6b4/41598_2024_76023_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f93/11470951/8992380d401d/41598_2024_76023_Fig5_HTML.jpg

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Hepatitis B virus genotypes in precision medicine of hepatitis B-related hepatocellular carcinoma: Where we are now.乙型肝炎病毒基因型在乙型肝炎相关肝细胞癌精准医学中的应用现状
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Single-cell landscape identifies the immunophenotypes and microenvironments of HBV-positive and HBV-negative liver cancer.单细胞图谱鉴定 HBV 阳性和 HBV 阴性肝癌的免疫表型和微环境。
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Clinical isolates of hepatitis B virus genotype C have higher in vitro transmission efficiency than genotype B isolates.乙型肝炎病毒 C 基因型的临床分离株比 B 基因型的分离株具有更高的体外传播效率。
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