猫疱疹病毒Ⅰ型(FHV-1)抗体ELISA检测方法的建立与初步应用

目的建立猫疱疹病毒Ⅰ型(FHV-1)抗体ELISA检测方法,应用于临床样品中FHV-1抗体的检测。方法培养猫肾传代细胞(CRFK),接种FHV-1病毒,制备CRFK正常抗原和FHV-1特异抗原,滴定山羊抗猫IgG-HRP和抗原最佳工作浓度,并进行特异性、敏感性、稳定性及精密性实验。结果正常抗原、特异抗原和酶结合物最佳工作浓度分别为0.05μg/mL、10μg/mL和1∶5 000;正常抗原、特异抗原批内变异系数分别为8.7%和5.8%,批间平均变异系数分别为11.9%和6.6%;检测灵敏度为1∶6 400;与猫细小病毒(FPV)、猫杯状病毒(FCV)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,可用于猫FHV-1抗体的检测。

Establishment and Preliminary Application of ELISA for Detecting Antibody to Felid Herpesvirus 1 in Cat

Objective To establish a ELISA method for detection of Felid herpesvirus 1 （ FHV-1 ） antibody in cat. Method Raises the CRFK cell, vaccinate the FHV-1 virus, prepare the normal CRFK antigen and special FHV- 1 antigen, titrate the best working density of enzyme union and the normal or special antigen and carry out the experiments of accuracy, sensitivity, stability and specificity. Result The best working density of normal, special antigen and the enzyme union is O. 05 ~g/mL, 10 Ixg/mL and 1：5 000 respectively; The inter-assay coefficient of variation of normal antigen and special antigen is 8.7% and 5.8% respectively, the intra-assay average coefficient of variation is 11.9% and 6.6% respectively; The detection sensitivity is 1：6 400. There is no cross-reactivity with Feline panleukopenia virus （FPV） and Feline calicivirus virus （FCV）. 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 can be used in detecting the Cat FHV-1 immune body.