Quantitative detection of serum procollagen Ⅲ N-terminal peptide chemiluminescence enzyme immunoassay

VernacularTitle:Ⅲ型前胶原氨基端肽的化学发光定量检测
Keywords: Chemiluminescence; Procollagen; Liver cirrhosis; Enzyme-linked immunoserbent assay
From: Chinese Journal of Experimental and Clinical Virology 2013;27(5):385-387
CountryChina
Language:Chinese
Abstract: Objective To establish chemiluminescence enzyme immunoassay (CLEIA) for quantitative detection of procollagen Ⅲ N-terminal peptide (P ⅢNP) in serum.Methods A sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of P Ⅲ NP as the catalytic enzyme and the luminol as the luminescence reagent.Several reactions liquid's concentration and reaction conditions were optimized.The method was evaluated in all aspects such as linear range,sensitivity,specificity,stability and so on.The CLEIA was compared with imported ELISA kits,by detecting clinical serum.Results The linear range was 0.8-85 ng/ml.The detection limit was 0.5 ng/ml.Inter-assay and intra-assay RSD were both less than 10％.The recoveries of three different spiked concentration samples were 96.2％,91.2％ and 101.1％.After stored at 4℃ and 37℃ for 3,5,7 days,the analysis showed correlation coefficient higher than 0.99 and RSD lower than 6％.The detected results of clinical sera with CLEIA closely corresponded to those with imported ELISA.Conclusion Established CLEIA for quantity determination of serum PⅢ NP has high accuracy,sensitivity and repeatability.