长爪沙鼠仙台病毒( SeV) 抗体ELISA 检测方法的建立与应用

摘要: 目的建立长爪沙鼠仙台病毒( SeV) 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SeV，制备鸡胚尿囊液正常抗原和SeV 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. 1 μg /mL、2 μg /mL 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SeV 抗体的检测。

Establishment and Preliminary Application of ELISA in Detecting Sendai Virus Antibody in Mongolian Gerbil

Abstract: 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 μg /mL，2 μg /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 TheELISA method is good in specificity，sensitivity，duplication，and stability，as a result of which ELISA can be used in detecting the Sendai virus antibody．