荧光抗体技术诊断犬瘟热的研究

Application of IFA Technique in Diagnosis of Canine Distemper

本研究建立了用于检测犬瘟热病毒的间接荧光抗体染色法。试验确定了1∶80稀释的抗犬瘟热单克隆抗体和1∶32倍稀释的荧光素标记羊抗鼠IgG为适宜的工作浓度。采用该方法检测21只犬瘟热自然感染犬的血液样品 ,CDV检出率为76%。对3只犬细小病毒自然感染犬 ,6只健康犬进行检测 ,各有一例为CDV抗原阳性。此外 ,用该方法检测了此21只犬瘟热病犬的眼分泌物、鼻汁、粪便和血涂片 ,其检出率分别为21%、14 %、15%和76 %。该方法与临床诊断的符合率为77%。另外 ,对稀释荧光素标记抗体冷冻保存 ,测定其保存期至少为7个月 ,染色后荧光稳定性为2周。该方法是一种简便、快速、特异的检测犬瘟热的方法 ,适于推广使用。

The indirect fluorescence antibody used to diagnose canine distemper(CD)was established.The suitable work concentration of monoclonal antibody against canine distemper(CD)was1:80and that of sheep-anti-mouse IgG-FITC was1:32.The white blood cells from21dogs with natural infection were detected by the IFA,yielding a76%positive rate and6healthy dogs without infection showing2positive ones.Among21dogs with CDV infecˉtion,21%of eye discharges,14%of nose discharges,14%of faecal samples and76%of blood smears showed positive results.Storage of diluted monoclonal antibody against canine distemper and smears dyed with sheep-anti-mouse IgG-FITC for7and2months respectively resulted in no marked change in fluorescence under fluorescent microscope.It is suggested that the established IFA is a rapid?simple and specific method for diagnosis of canine distemper.