摘要
目的建立长爪沙鼠仙台病毒(Se V)抗体ELISA检测方法,用于长爪沙鼠体内仙台病毒抗体的检测。方法孵育鸡胚,接种Se V,制备鸡胚尿囊液正常抗原和Se V特异抗原,滴定酶结合物和抗原最佳工作浓度,并进行特异性、敏感性、精密性、稳定性实验。结果正常抗原、特异抗原和酶结合物最佳工作浓度分别为0.1μg/m L、2μg/m L和1∶5 000;正常抗原、特异抗原批内变异系数分别为9.6%和8.4%,批间平均变异系数分别为9.0%和5.9%;检测灵敏度为1∶10 240;与沙鼠小鼠肝炎病毒(MHV)、小鼠肺炎(PVM)、呼肠孤病毒3型(Reo3)均无交叉反应。稳定性试验相对偏差小于25%。结论建立的ELISA方法特异性、敏感性强,重复性、稳定性好,检测结果,准确、可靠。可用于长爪沙鼠体内Se V抗体的检测。
Objective To develop a ELISA method for determination of Sendai virus（SeV） antibody in Mongolian gerbil. Methods Hatchs chicken embryos, vaccinate the Sendai Virus, prepare the normal chicken embryo allantoic fluid antigen and special SeV antigen, titrate the best working density of enzyme union and the normal or special antigen and carry out the experiments of specificity, sensitivity, accuracy and stability. Result The best working density of normal, special antigen and the enzyme union is 0. 1 I^g/mL, 2 I^g/mL and 1 ： 5 000 respectively; The inter-assay coefficient of variation of normal antigen and special antigen is 9.6% and 8.4% respectively, the intra-assay average coefficient of variation is 9.0% and 5.9% respectively; The detection sensitivity is 1：10 240; There is no cross-reactivity with mouse hepatitis virus （MHV） , Pneumonia virus of mice （PVM）, and Reovirus type 3. The stability test shows the relative deviation is below 25%. Conclusion The ELISA method is good in specificity, sensitivity, duplication, and stability, as a result of which ELISA can be used in detecting the Sendai virus antibody.
出处
《实验动物科学》
2015年第6期39-43,共5页
Laboratory Animal Science
基金
国家科技支撑计划(No:2015BAI09B00)
