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Robust in vitro assay for analyzing the neutralization activity of serum specimens against hepatitis B virus.


ABSTRACT: Anti-HBs is a well-known marker of protective capability against HBV. However, little is known about the association between the qAnti-HBs determined by immunoassays and the neutralization activity (NAT) derived from functional assays. We developed an in vitro assay for direct measurement of the NAT of human sera. The new assay was highly sensitive, with an analytical sensitivity of 9.6 ± 1.3 mIU/mL for the HBIG standard. For serum detection, the maximum fold dilution required to produce ?50% inhibition (MDF50) of HBV infection was used as the quantitative index. In vitro NAT evaluations were conducted for a cohort of 164 HBV-free healthy individuals. The results demonstrated that the NAT positively correlated with the qAnti-HBs (R2?=?0.473, p?50???8) was 62.9 mIU/mL, with an AUROC of 0.920. Additionally, we found that the qAnti-HBc was another independent parameter positively associated with the NAT (R2?=?0.300, p?Abbreviations: HBV: Hepatitis B virus; HBsAg: Hepatitis B surface antigen; Anti-HBs: Hepatitis B surface antibody; HBeAg: Hepatitis B e antigen; Anti-HBc: Hepatitis B core antibody; qAnti-HBs: quantitative hepatitis B surface antibody; qAnti-HBc: quantitative hepatitis B core antibody; qHBeAg: quantitative hepatitis B e antigen; NAT: neutralization activity; HBIG: hepatitis B immune globulin; NTCP: Na+-taurocholate cotransporting polypeptide; IRES: internal ribosome entry site; ccHBV: cell culture derived hepatitis B virus; GE/cell: genome equivalent per cell; MOI: multiplicity of infection; Dpi: day post infection; HepG2-TetOn: a HepG2-derived cell line that expresses the doxycycline-regulated transactivator; ROC: receiver operating characteristic curve; AUROC: area under receiver operating characteristic curve; LLOQ: the lower limits of quantification; MDF50: the maximum fold dilution required to produce ?50% inhibition; IC50: half maximal inhibitory concentration.

SUBMITTER: Zhang YL 

PROVIDER: S-EPMC6542156 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Robust <i>in vitro</i> assay for analyzing the neutralization activity of serum specimens against hepatitis B virus.

Zhang Ya-Li YL   Gao Ying Y   Cao Jia-Li JL   Zhao Jing-Hua JH   Zhang Tian-Ying TY   Yang Chuan-Lai CL   Xiong Hua-Long HL   Wang Ying-Bin YB   Ou Shan-Hai SH   Cheng Tong T   Chen Chang-Rong CR   Yuan Quan Q   Xia Ning-Shao NS  

Emerging microbes & infections 20190101 1


Anti-HBs is a well-known marker of protective capability against HBV. However, little is known about the association between the qAnti-HBs determined by immunoassays and the neutralization activity (NAT) derived from functional assays. We developed an <i>in vitro</i> assay for direct measurement of the NAT of human sera. The new assay was highly sensitive, with an analytical sensitivity of 9.6 ± 1.3 mIU/mL for the HBIG standard. For serum detection, the maximum fold dilution required to produce  ...[more]

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