Development of quantitative detection kit for 25- hydroxy vitamin D (chemiluminescence)

• 1、National Engineering Laboratory for Cereal Fermentation Technology, the Key Laboratory of Industrial Biotechnology, Ministry of Education, the Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122

Abstract：To develop and evaluate a chemiluminescent immunoassay (CLIA) test kit for quantitative determination of 25-hydroxyvitamin D（25-OHD）in serum. Methods The 96 well microplate was coated with 25-hydroxyvitamin D monoclonal antibodies, HRP conjugate( the mixture of biotinylated 25-OHD and streptavidin-labeled HRP ) compete with the 25-OHD released from sample to bind sites of antibodies. And evaluate the kit. Results The minimum detection limit of the kit was 4.90 ng/mL, the bias was less than 10％, the intra-assay and inter-assay coefficients of variation were 6.5%, and 8.9%. Its preserving duration could be 12 months at 2-8℃. 200 clinical serum specimens were detected with this method and a foreign serum ferritin kit (Roche), the correlation coefficient was 0.9717. Conclusion Successfully prepared 25-hydroxy vitamin D (CLIA) quantification kit .And all the technical parameters reached the industrial standards. The kit was sensitive, stable, fast and simple.

Keywords： 25-hydroxyvitamin D chemiluminescent immunoassay kit