State Key Laboratory of Cancer Biology, Department of Pharmacogenomics, School of Pharmacy, The Fourth Military Medical University, 169 West Changle Road, Xi'an, 710032, China.
Eur J Clin Microbiol Infect Dis. 2013 Dec;32(12):1557-64. doi: 10.1007/s10096-013-1910-0. Epub 2013 Jun 19.
The quantitative measurement of serum hepatitis B surface antigen (HBsAg) is important for monitoring anti-hepatitis B virus (HBV) therapy and evaluating the safety of blood products or blood transfusion. This study is aimed at generating and evaluating a capillary-based chemiluminescence immunoassay for the simple and rapid detection of human serum HBsAg. A simple and rapid capillary chemiluminescence immunoassay (CCIA) was developed using a portable analyzer for quantitatively detecting the levels of HBsAg in human serum. The experimental conditions were optimized, and the sensitivity and specificity of the CCIA were validated in positive HBsAg (HBsAg(+)) and negative HBsAg (HBsAg(-)) human sera. The CCIA quantitatively detected the levels of human serum HBsAg at 0.4-15.0 ng/mL, which resulted in dose-dependent increases in the chemiluminescence (CL) signals, with a sensitivity of 0.3 ng/mL. The assay took only 25 min for the analysis of a single sample. The CCIA only detected HBsAg, but not hepatitis C virus (HCV), human immunodeficiency virus-1 (HIV-1) p24, Treponema (TP) antigens, or other serum proteins and lipids tested with a specificity of 100%. The CCIA for detecting HBsAg was reproducible, with a low intra-assay coefficient of variation (CV) (6.7%) and inter-assay CV (7.4%). The validation of 280 known HBsAg(+) or HBsAg(-) clinical serum samples revealed that the CCIA detected 97.5% of HBsAg(+) sera without false-positives, with a sensitivity and specificity similar to that of the Architect i2000SR HBsAg assay (Kappa value = 0.983). The CCIA is a simple and rapid method for quantitatively measuring the levels of serum HBsAg to monitor anti-HBV therapy and evaluate the safety of blood samples for transfusion.
本研究旨在开发一种基于毛细管的化学发光免疫分析方法，用于简单快速地检测人血清乙型肝炎表面抗原（HBsAg）。 我们使用便携式分析仪开发了一种简单快速的毛细管化学发光免疫分析（CCIA），用于定量检测人血清中 HBsAg 的水平。 优化了实验条件，并在阳性 HBsAg（HBsAg(+)）和阴性 HBsAg（HBsAg(-)）人血清中验证了 CCIA 的灵敏度和特异性。 CCIA 在 0.4-15.0ng/mL 范围内定量检测人血清 HBsAg 的水平，导致化学发光（CL）信号呈剂量依赖性增加，灵敏度为 0.3ng/mL。 单次样品分析仅需 25 分钟。 CCIA 仅检测 HBsAg，而不检测丙型肝炎病毒（HCV）、人类免疫缺陷病毒-1（HIV-1）p24、梅毒螺旋体（TP）抗原或其他经测试具有 100%特异性的血清蛋白和脂质。 用于检测 HBsAg 的 CCIA 具有可重复性，日内变异系数（CV）（6.7%）和日间 CV（7.4%）均较低。 对 280 份已知 HBsAg(+)或 HBsAg(-)临床血清样本的验证表明，CCIA 检测到 97.5%的 HBsAg(+)血清无假阳性，灵敏度和特异性与 Architect i2000SR HBsAg 检测（Kappa 值=0.983）相似。 CCIA 是一种简单快速的方法，用于定量测量血清 HBsAg 水平，以监测抗 HBV 治疗并评估输血样本的安全性。
