摘要
目的建立长爪沙鼠小鼠肝炎病毒（MHV）抗体ELISA检测方法。方法培养DBT细胞，接种MHV-V1、MHV-V3、MHV-JHM病毒，制备DBT正常抗原和MHV-V1、MHV-V3、MHV-JHM三价特异抗原，滴定酶结合物和抗原最佳工作浓度，并进行精密性、敏感性、稳定性、特异性实验。结果正常抗原、特异抗原和酶结合物最佳工作浓度分别为0．4gg／ml、5gg／ml和1：5000；正常抗原、特异抗原批内变异系数分别为7．9％和6．8％，批间平均变异系数分别为12．7％和11．2％；检测灵敏度为1：1280；与小鼠仙台病毒（sV）、小鼠淋巴细胞脉络丛脑膜炎病毒（LCMV）均无交叉反应。稳定性试验相对偏差小于25％。结论建立的ELISA方法重复性、稳定性好，特异性、敏感性强。可用于沙鼠MHV抗体的检测。
Objective To develop a ELISA method for determination of Mouse Hepatitis Virus antibody in Mongolian gerbils. Methods The cultured DBT cell were vaccinated with MHV-V1, MHV- V3, MHV-JHM virus for preparation the normal DBT antigen and special MHV antigen, the best working density of enzyme union and the normal or special antigen were titrated and the experi- ments of accuracy, sensitivity, stability and specificity were done. Result The best working density of normal, special antigen and the enzyme union is 0.4 μg/ml, 5μg/ml and 1 ： 5000 respectively; The inter- assay coefficient of variation of normal antigen and special antigen is 7.9% and 6.8% respectively, the intra-assay average coefficient of variation is 12.7% and 9.7% respectively; The detection sensitivity is 1：1280; There is no cross-reactivity with Sendal virus （SV） and Lymphocytic Choriomeningitis virus （LCMV）. The stability test shows the relative deviation is below 25%. Conclusion The ELISA method is good in duplication, stability, specificity, and sensitivity, as a result of which ELISA may be used in detecting the antibody to MHV in Mongolian gerbil.
出处
《实验动物与比较医学》
CAS
2013年第3期204-209,共6页
Laboratory Animal and Comparative Medicine
基金
实验动物新品种的种群建立与质量标准化研究（国家科技支撑计划项目：2011BA115801）
