CAJ | 학술논문

采用RT-PCR方法直接从芦花鸡肿瘤中进行禽白血病病毒（ Avian leucosis virus， ALV）的核酸扩增，经测序证实为ALV。将上述病料接种DF1细胞（ C/E）进行盲传，并对其感染细胞上清进行ALV P27抗原检测，结果为阳性，证实获得了一株病毒，命名为SDJN2012。用PCR 方法扩增其囊膜蛋白gp85基因并测序，与GenBank中的ALV参考毒株进行核苷酸或氨基酸同源性比较。结果表明：SDJN2012与山东较早发现的J亚群ALV代表株SD0001的核苷酸（氨基酸）同源性最高，达93.4％（92.6％），与J亚群不同年代的代表株同源性为88.6％～93.3％（88.2％～90.6％），而与 A， C，D， E 亚群 ALV 毒株的同源性仅为19.7％～32.2％（10.2％～32.2％），进一步确证分离到的病毒为J亚群ALV。对种蛋的跟踪监测证实：ALV蛋清P27抗原阳性率为37.6％，而血清学监测则呈现多样性的变化：鸡群发病前J亚群ALV抗体检测为阴性，发病后阳性率仅为9.35％，而A/B亚群阳性率则由发病前的9.8％，攀升到发病后的75.4％，彰显出ALV病原学、血清学和分子流行病学的复杂性。
채용RT-PCR방법직접종호화계종류중진행금백혈병병독（ Avian leucosis virus， ALV）적핵산확증，경측서증실위ALV。장상술병료접충DF1세포（ C/E）진행맹전，병대기감염세포상청진행ALV P27항원검측，결과위양성，증실획득료일주병독，명명위SDJN2012。용PCR 방법확증기낭막단백gp85기인병측서，여GenBank중적ALV삼고독주진행핵감산혹안기산동원성비교。결과표명：SDJN2012여산동교조발현적J아군ALV대표주SD0001적핵감산（안기산）동원성최고，체93.4％（92.6％），여J아군불동년대적대표주동원성위88.6％～93.3％（88.2％～90.6％），이여 A， C，D， E 아군 ALV 독주적동원성부위19.7％～32.2％（10.2％～32.2％），진일보학증분리도적병독위J아군ALV。대충단적근종감측증실：ALV단청P27항원양성솔위37.6％，이혈청학감측칙정현다양성적변화：계군발병전J아군ALV항체검측위음성，발병후양성솔부위9.35％，이A/B아군양성솔칙유발병전적9.8％，반승도발병후적75.4％，창현출ALV병원학、혈청학화분자류행병학적복잡성。
Avian leukosis virus (ALV) was isolated from the tumors of Chinese native breed “Luhua” chicken, and was confirmed by RT-PCR and DNA sequencing .Then, a virus strain, named SDJN 2012, was confirmed by detec-tion of P27 antigen from the virus supernatant via inoculation of DF 1 cells ( C/E) .The gene of envelope protein gp 85 were cloned and sequenced by designing specific primers of pg85 motif from ALV reference strains in GenBank , and the nucleotide and amino acid homologies were compared .The results showed that the SDJN 2012 had the highest nu-cleotide ( amino acid) homology93.4%(92.6%) with SD0001 strain, the first viruses of the subtype J strains dis -covered in Shandong .Besides, SDJN2012 shared 88.6%~93.3% ( 88.2%~90.6%) nucleotide ( amino acid ) homologies with different subtypes of J strains , and 19.7%~32.2%(10.2%~32.2%) homologies of A , C, D, E subtypes of ALV homolog , which further confirmed that the isolated virus was J subtype ALV .The positive rate of P27 antigen was 37.6%.The antibodies of ALV J subtype were negative before onset , and were only 9.35%positive after onset.While, the positive rate of A/B subtype from climbed to 75.4%after onset from 9.8%, which indicated the complexity of ALV clinical etiology , serology and molecular epidemiology .