【摘要】 本研究对1例具有剧烈搔痒症状,但无明确传播途径的病犬的组织进行伪狂犬病毒(Pseudorabies virus,PRV)特异性PCR检测,从脑干、三叉神经、交感神经和颈段脊髓中扩增出预期大小的DNA片段。将PCR为阳性的脑干组织病料接种Vero细胞,培养72 h后细胞发生圆缩、脱落等病变。对出现稳定病变的细胞培养物进行PRV PCR检测获得预期大小的DNA片段,用PRV mAb进行免疫组化染色,部分细胞中出现大量棕褐色阳性颗粒,从而判定感染细胞中存在PRV,将此分离株命名为BJ/RD株。根据GenBank登录的PRV(JF797219)序列设计引物,扩增gE基因,并对扩增产物进行序列测定与分析,结果显示,BJ/RD株与此前分离的犬源毒株BJ/YT及2012年河北猪群分离株HB/HD的gE基因序列完全相同(核酸同源性为100%),与2012年河北地区分离株HB/HS、HB/BD、HB/LF核苷酸同源性99.9%,而与国内外其他猪源PRV分离株gE基因序列有一定差异。更多还原

【Abstract】 Typical pseudorabies-like itching sign was observed in a dog that had no clear route of transmission.Samples of sympathetic trunk,cervical spinal cord,trigeminal nerve and brainstem were tested using PRV-specific PCR.The PRV DNA fragments were detected in these tissues.Then brainstem was homogenized and inoculated into Vero cell cultures.At 72 h post inoculation,PRV specific CPE was showed in Vero cells,and the results of immunohistochemical test and PCR were positive.The results indicated that the isolated virus was PRV and designated as BJ/RD strain.The gE gene of the BJ/RD strain was sequenced and compared with the known sequences in GenBank.The BJ/RD strain shared 100% nucleotide homology with canine BJ/YT strain from Beijing and porcine HB/HD strain from Hebei province,and 99.9% nucleotide homology with HB/HS,HB/BD and HB/LF strains.更多还原