中国人兽共患病学报2017，Vol.33Issue(12)：1060-1065,6.DOI:10.3969/j.issn.1002-2694.2017.12.002
牛结核γ-干扰素夹心ELISA检测试剂盒的研制与评价
Development and evaluation of gamma-interferon sandwich ELISA kit in herds infected with Mycobacterium bovis
摘要
Abstract
This study was aimed to develop a sandwich ELISA kit for the diagnosis of bovine tuberculosis.And it was applied and evaluated in the quarantine of bovine tuberculosis.We established a bovine IFN-γ release method in vitro and developing three batches of kits.The sensitivity,repeatability and retention period of the kit were all evaluated.Totally 961 serum samples were tested using the developed sandwich ELISA kit tuberculin skin test and a commercial ELISA kit.Our results showed that the detection limit of this ELISA was 8.21 mg/mL.The repeatability tests showed good reproducibility in the intraassay and inter-assay.At the same time,the retention period of the kit was more than 12 months.Compared with the tuberculin skin test,the positive coincidence rate was 70.59％ and the negative coincidence rate was 99.20％,while the total coincidence rate was 98.44％.And compared with the BOVIGAMTM kit,the positive coincidence rate was 91.30％ and the negative coincidence rate was 99.78％,while the total coincidence rate reached 99.58％.At the same time,the sensitivity and specificity of the sandwich kit were 85.00％ and 100％,respectively.We established a bovine IFN-γ release method in vitro and developing corresponding kits successfully have a good application prospect.关键词
牛结核病/γ-干扰素/夹心ELISA/试剂盒/敏感性/特异性Key words
bovine tuberculosis/IFN-γ/sandwich ELISA/kit/specificity/sensitivity分类
医药卫生引用本文复制引用
李昕,焦新安,徐正中,单法,夏爱鸿,孟闯,沈也驰,陈义平,南文龙,陈祥..牛结核γ-干扰素夹心ELISA检测试剂盒的研制与评价[J].中国人兽共患病学报,2017,33(12):1060-1065,6.基金项目
国家重点研发计划(No.2017YFD0500300) （No.2017YFD0500300）
国家自然科学基金(No.31602031) （No.31602031）
江苏科技计划项目(No.BE2015343,No.BK20160466,No.BK20171285,No.16KJB230003) （No.BE2015343,No.BK20160466,No.BK20171285,No.16KJB230003）
江苏‘青蓝工程’‘六大人才高峰’(No.SWYY-083)和优势学科建设工程项目联合资助 Funded by the National Key Research and Development Program of China (No.2017YFD0500300),the National Natural Science Foundation of China (No.31602031),the Science and Technology Program of Jiangsu (Nos.BE2015343,BK20160466,BK20171285 and 16KJB230003),the Qinglan Project,Six Talent Peaks Project (No.SWYY-083),and the Priority Academic Development Program of Jiangsu Higher Education Institutions (PADP). （No.SWYY-083）
