期刊
CLINICA CHIMICA ACTA
卷 426, 期 -, 页码 139-144出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.cca.2013.09.013
关键词
Amplified luminescent proximity homogeneous assay; Chemiluminescence; Hepatitis B surface antigen; Immunoassays
资金
- National Key Science and Technology Program [2009ZX10004-706]
- National Natural Science Foundation of China [81271931]
- Natural Science Foundation of Guangdong Province [S2012010009547]
Background: Hepatitis B virus (HBV) poses a serious risk to human health and the hepatitis B surface antigen (HBsAg) is a popular biomarker in the diagnosis of HBV infection. A quantitative method with a high degree of accuracy, sensitivity and throughput is needed. Methods: A novel amplified luminescent proximity homogeneous assay (AlphaLISA) was developed for HBsAg determination. A set of monoclonal antibodies was screened against the main subtypes of HBsAg (adr, ay) to confirm the assay's sensitivity to mutants. Technological processes and reaction conditions were optimized and the assay performance was evaluated. Results: HBsAg concentrations were determined within a linear range of 0.04 to 100 IU ml(-1). The detection sensitivity was established as 0.01 IU ml(-1). Assay sensitivity to mutant HBsAg was achieved through antibody screening. The results demonstrate that the reproducibility, recovery, and specificity of this assay for HBsAg were better than acceptable. Compared with the commercial light-initiated chemiluminescence assay, the correlation coefficient of the novel assay was established as 0.921. Conclusions: The novel AlphaLISA developed in this study has shorter incubation time and easier protocol than the ones of conventional ELISA. It could be used for the clinical determination of HBsAg in human serum. We established a platform for further development of other biomarkers. (C) 2013 Elsevier B.V. All rights reserved.
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