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)