Establishment and application of light initiated chemiluminescence assay of high mobility group box 1 / 中华核医学与分子影像杂志

ABSTRACT

Objective To establish a fast and quantitative light initiated chemiluminescent assay (LICA) method for high mobility group box1 (HMGB1) determination.Methods Two strains of paired HMGB 1 monoclonal antibodies were used.One was used to coat receptor microspheres.The other was labeled with biotin first and then composed with chain mildew element of affinity donor microsphere to form LICA method for HMGB1.After optimizing the reaction system,the technical specifications of the method was evaluated.Serum HMGB1 levels of common pneumonia patients (CPP) and severe pneumonia patients (SPP) were measured and compared with that of health controls.Two-sample t test was used.Results The sensitivity of LICA was 0.1 μg/L,with linear measurement ranging from 0.1 to 1 000 μg/L.The precisions of intra-and inter-analysis were 1.74％-2.92％ and 1.93％-3.73％ respectively,both were lower than 5％.The recovery rate was 99.74％ (range94.53％-106.37％).The correlation coefficient of LICA and enzyme-linked immunosorbent assay (ELISA) was 0.888 2.The LICA method had good specificity and no obvious cross reaction with HMGB2 and HMGB3.The serum HMGB1 level in CPP (n=35) and SPP (n=25) was significantly higher than that in health controls (n=35)(6.76±3.13),(19.69±+9.04) vs (1.49±+0.74) μg/L;t values-5.447 and-5.186,both P＜0.01.The HMGB1 levels between CPP and SPP were also significantly different (t=-3.500,P＜0.01).Conclusions The established LICA method of HMGB1 has high sensitivity and specificity with reliable results.This method is also homogeneous,fast and cleaning-free,thus has a good prospect in clinical application.