犬瘟热的微生物学诊断研究

Microbiological Diagnosis of Canine Distemper

对临床诊断为犬瘟热的１４例病犬，取其脑组织，运用细胞培养、合胞体检查、包涵体检查、免疫荧光试验、电镜观察等５种检测方法，就犬瘟热的微生物学诊断进行了系统的研究。结果表明，犬脑组织块与猫胚（ＦＥ）细胞或非洲绿猴肾（Ｖｅｒｏ）细胞共同培养，盲传的培养物出现不稳定的细胞病变，通过对不同代次的培养物检查包涵体，并作超薄切片或负染，电镜观察犬瘟热病毒（ＣＤＶ）粒子，证实分离到ＣＤＶ野毒。病犬脑组织切片经ＨＥ染色检查包涵体及用ＣＤＶ荧光抗体直接法检测脑抹片及接种犬脑的细胞培养物，均获得一定的阳性结果。建立的改良离子捕获电镜法，用于检测犬脑匀浆和犬脑接种细胞培养物，与离子捕获电镜法、免疫电镜法及直接电镜法相比，效果更为理想。１４例临床诊断为犬瘟热的病犬，综合运用上述微生物学手段检测，结果为１２例阳性，２例疑似。

Fourteen dogs clinically suspected of canine distemper(CD)in Nanjing were examined microbiologically.Cytopathic effects(CPE)were observed after co cultivation of the brain tissue cul tures with Vero cells or FE cells.ACDV field strain has been isolated by blind passage in FE and Vero cells.This isolate was demonstrated by electron microscopy(EM)both in thin sections and neg ative stained specimens from cell cultures.Paraffin sections of the brains and lungs were made for ex amination of inclusion bodies.CDV direct immunofluorecent technique(IF)was also applied to detect the brain smears and the cell cultures.7of12specimens are positive for inclusion bodies and in6of7specimens CDV are demonstrated by IF.Amodified ion capture,which combined with ion capture EM,immuno electron microscopy(IEM)and adding protease inhibitor PMSF to the specimens,was established for investigation of CDV.Conclusively,with different microbiological methods,12of14dogs were diagnosed as CDV positive,2of14dogs were suspective for CD.