猪传染性胃肠炎免疫胶体金诊断试剂条的研制

Development and Preliminary Application of Rapid Test Strip for Porcine Transmissible Gastroenteritis Virus

采用柠檬酸钠还原法制备胶体金颗粒,用TGEV单克隆抗体标记,并包被在玻璃纤维膜上。另外将纯化的TGEV单克隆抗体和羊抗小鼠抗体包被在硝酸纤维膜上,组装成TGEV快速检测条,对其灵敏性、特异性、稳定性及保存期检测。结果表明,研制的TGEV快速检测条检出TGEV最低量为150μg/mL,只能检测出TGEV,而与猪瘟病毒培养液、猪繁殖障碍与呼吸综合症(PRRS)、鸡减蛋综合症(EDS-76)病毒、鸽新城疫(PPMV-1-PL)病毒、大肠杆菌及沙门氏菌无交叉反应;研制的三批试验条性能稳定,保存期试验证实该条在常温下可保存1年。试验证实所研制的试剂条具有灵敏特异稳定无交叉反应,可用于生产实际。

A kind of method for determining porcine transmissible gastroenteritis virus （TGEV） by using colloidal gold im- munochromatography assay （GICA） was developed. Using sodium citrate reduction method preparing colloidal gold particles, and purified by TGEV monoclonal antibody labeling, and in glass fiber membrane, and the purified TGEV monoclonal antibody and Goat anti mouse antibody coated on nitrocellulose membrane, assembly of TGEV rapid detection, the sensitivity, specifici- ty, stability and retention period detection, and compared the test. The results showed that the development of rapid detection, TGEV detection of TGEV minimum quantity is 150 tzg/mL; only TGEV can be detected, and the culture liquid, porcine repro- ductive and respiratory syndrome and classical swine fever virus （PRRS）, egg drop syndrome virus （EDS--76）, pigeon New- castle disease virus （PPMV--1--PL）, Escherichia coli and Salmonella no cross reaction∽ developed the three batch of the test strip and stable performance; preservation test confirmed the stored at room temperature for 1 years; With South Korea made in article reagent contrast tests of different samples, the results are consistent. Which confirmed the developed reagent sensitive specific stability without cross reaction, can be used in actual production.