地方品种HR土鸡不同亚型禽白血病病毒的共感染

Co-infection status of different subgroups of avian leukosis virus in HR local layer chicken

为研究HR土鸡中存在的不同亚型禽白血病病毒(ALV)共感染的情况,本实验分别采集46只HR土公鸡的泄殖腔棉拭子和455枚鸡蛋卵白样品,采用ELISA试剂盒检测p27抗原;并采用相应的ELISA试剂盒分别检测卵黄中J亚型ALV(ALV-J)和AB亚型ALV(ALV-AB)抗体。结果表明:HR土公鸡泄殖腔棉拭子样品中p27检出阳性率为87%(40/46),卵白检出率为74.7%(340/455);而卵黄中ALV-J和ALV-AB抗体阳性率分别为0(0/30)和80%(24/30)。无菌采集初步筛选p27抗原检测为阳性的5只HR土公鸡的抗凝血接种CEF,采用抗ALV-J和ALV-A的单克隆抗体进行IFA检测,结果显示5份样品中ALV-A和ALV-J的阳性率均为100%(5/5)。同时选取HR土鸡分离株HR332进行PCR扩增鉴定,结果表明分离株HR332存在ALV-J(HR332J)和ALV-A(HR332A)。其中,HR332J与11株ALV-J国内外参考株的同源性为92.4%～97.9%;HR332A与ALV-A参考株RSA-A、MQNCSU的同源性分别为90.1%和89.7%,与国内分离株SDAU09E2的同源性为99.0%。本研究显示,地方品种HR土鸡存在不同亚型ALV共感染,同时ALV-A和ALV-J共感染同一个鸡的现象已经存在。

Abstract： In the present study, the co-infection of avian leukosis vires （ALV） was investigated in HR local layer chicken flock. Fourty-six cloacal swabs and 455 egg whites were collected for p27 antigen detection, while 30 egg yolk were collected for detection of ALV-AB and ALV-J antibody by ELISA. The results showed that the p27 positive rate of cloacal swabs and egg whites were 87% （40/46） and 74.7% （340/455）; the antibody against ALV-AB and ALV-J in egg yolk were 0 （0/30） and 80% （24/30）, respectively. ALV-A and ALV-J were isolated in cell culture and identified by immunofluorescence assay from the 5 random selected p27 positive HR local layer chickens. Of which the 924 bp （ALV-J） and 1,127 bp （ALV-A） were amplified from virus isolates of HR332J and HR332A with 2 specific pairs of primers of ALV-J and ALV-A by PCR, respectively. The sequence analysis of ALV-J gp85 gene （924 bp） indicated that the homology between isolate HR332J and 11 reference strains in GenBankwas 92.4% to 97.9%; the isolate HR332A and reference strains of RSA-A, MQNCSU were 89.7% and 90.1% respectively, but up to 99.0% homology with domestic strain SDAU09E2. The study showed that co-infection of different subgroups of ALVs existed in HR local layer chickens.