CAJ | 학술논문

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。
본연구대1례구유극렬소양증상，단무명학전파도경적병견적조직진행위광견병독（Pseudorabies virus，PRV）특이성PCR검측，종뇌간、삼차신경、교감신경화경단척수중확증출예기대소적DNA편단。장PCR위양성적뇌간조직병료접충Vero세포，배양72 h후세포발생원축、탈락등병변。대출현은정병변적세포배양물진행PRV PCR검측획득예기대소적DNA편단，용PRV mAb진행면역조화염색，부분세포중출현대량종갈색양성과립，종이판정감염세포중존재PRV，장차분리주명명위BJ/RD주。근거GenBank등록적PRV（JF797219）서렬설계인물，확증gE기인，병대확증산물진행서렬측정여분석，결과현시， BJ/RD주여차전분리적견원독주BJ/YT급2012년하북저군분리주HB/HD적gE기인서렬완전상동（핵산동원성위100%），여2012년하북지구분리주HB/HS、HB/BD、HB/LF핵감산동원성99.9%，이여국내외기타저원PRV분리주gE기인서렬유일정차이。
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.