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通过血清学、病理学以及病毒学检测,首次在国内从感染ALV-J的海蓝白商品蛋鸡群中分离到B亚群禽白血病病毒(ALV-B)。ELISA检测该鸡场血清样品(92份)显示,ALV-A/B型抗体阳性率为16.3%,ALV-J抗体阳性率为13%,其中有1只鸡呈现抗体双阳性。取3只表现临床症状病鸡做病理组织学观察,在肝脏同一位置发现淋巴细胞样瘤细胞和髓细胞样瘤细胞增生灶,其它组织器官(除脑、神经和法氏囊)也有不同程度的两种瘤细胞增生。根据文献设计合成ALV-A、ALV-B、ALV-J、MDV和REV的特异性引物。取该3只病鸡肝脏病料研磨接种DF-1细胞,培养7~10天后提取培养细胞DNA,PCR扩增获得1 100bp的ALV-B特异性片段和924bp的ALV-J特异性片段,其中有1只鸡为ALV-B和ALV-J双阳性,而ALV-A、MDV、REV的PCR结果为阴性,因此获得ALV-B和ALV-J各两株分离株。对ALV-B、ALV-J扩增序列进行遗传进化树分析表明,两ALV-B分离株与其代表株的同源性分别为92.8%和94.7%;两ALV-J分离株与英国原型株同源性分别为96.9%和91.5%,与美国原型株同源性为85.9%和81.5%,与国内分离毒株同源性在87.0%~98.7%之间。本研究在国内分离到ALV-B毒株,并证实了ALV-B、J在同一只鸡体内的混合感染,应引起养禽业及相关研究部门的高度重视。
Abstract：Two strains of Avian leukosis virus subgroup B(ALV-B) were isolated for the first time in China Hy-line White on the cultured DF-1 cells which were inoculated tissue samples from by an ELISA assay,a histopathology examination and a PCR-based diagnosis.The results from the ELISA assay indicated that the positive rate of serum antibodies to ALV-B and ALV-J virus were 16.3%(15/92) and 13%(12/92),respectively.The histopathological examination indicated that two types of tumor cells existed at same focus in liver and spleen,which mainly were myelocytoma cells and lymphosarcoma cells.The PCR-based diagnosis were performed as follows: the cellular DNA was extracted from the inoculated DF-1 cells;the specific fragments of 1100bp and 924bp were obtained by a PCR system with the diagnostic primers of ALV-B and ALV-J;and the PCR results for ALV-A,MDV and REV were all negative.Then,the amplified fragments of the two ALV-B stains were partially sequenced and shown an identity of 92.8%,94.7% with the prototype strain of ALV-B(RSV Schmidt-ruppin B).The identities of two ALV-J strains with the prototype strain HPRS-103 at 96.9%,91.5%;The identities of two ALV-J strains with the American prototype strain at 85.9%,81.5%.Our study had shown that ALV-B was isolated for the first time from the ALV-J infected commercial layer flocks in China.It also indicated that the chance of genetic recombination among various subgroups of ALV was increased.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.002038
中图分类号:S852.659.3
引用信息:
[1]刘功振,张洪海,刘青,等.感染J亚群禽白血病病毒的蛋鸡群中检测出B亚群禽白血病病毒[J].病毒学报,2009,25(06):445-451.DOI:10.13242/j.cnki.bingduxuebao.002038 .
基金信息:
国家自然科学基金(30600444); 山东省优秀中青年科学家奖励基金(2005BSB01156)
2009-11-15
2009-11-15