一株水貂犬瘟热病毒的分离与鉴定

Isolation and Identification of Ferret Canine Distemper Virus

为寻找免疫失败的原因,有效防治犬瘟热的流行,对临床一水貂犬瘟热疑似病例,取其肝、脾、脑等组织研磨后接种Vero和BHK细胞分离病毒,并对分离毒株进行一系列鉴定。通过电镜观察,发现了大小约150 nm的副黏病毒样粒子。结果表明,分离株对鹅、鸡、小鼠、家兔、山羊、猪红细胞均无凝集性,该分离株的毒价TCID50为10-4.87;分离株病毒对乙醚、氯仿敏感,病毒的核酸型为RNA。经间接免疫荧光试验,接种病毒的BHK细胞出现特异性的亮绿色荧光。对分离株和对照毒株分别提取RNA进行RT-PCR扩增,最后得到与预期扩增片段(294 bp)相符的核酸电泳带,充分证明分离病毒为犬瘟热病毒。

Tissues of liver, spleen and brain from a ferret clinically suspected of canine distemper were used for isolating virus on Vero and BHK cell lines. Typical morbillivirus particles with a diameter of 150 nm were observed under direct electron microscope. The isolate can＇ t agglutinate erythrocytes such as goose,, chicken, rat, rabbit, goat, swine and it＇s TCID50 was 10^4.87. The virus was composed of RNA and sensitive to hydroxybenzene and chloroform. The inoculated BHK cell lines showed specific green fluorescence by immuofluresecence assay. The nucleotide fragment of 294 bp was amplified as expected by RT-PCR using RNA extracted from the isolate and CDV strains as templates. The results also showed that this strain was ferret canine distemper virus.