Takiuchi E, Médici K C, Alfieri A F, Alfieri A A
Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, Centro de Ciências Agrárias, Universidade Estadual de Londrina (UEL), Londrina 86051-990, Paraná, Brazil.
Res Vet Sci. 2005 Aug;79(1):85-8. doi: 10.1016/j.rvsc.2004.11.005. Epub 2004 Dec 21.
Bovine herpesvirus type 1 (BHV-1) was investigated by a semi-nested polymerase chain reaction (SN-PCR) and by MDBK cell culture virus isolation in organ fragments from 55 aborted fetuses collected from beef and dairy cattle herds with history of reproductive problems in the North of Paraná State, Brazil. A 425 bp amplicon of the BHV-1 glycoprotein D gene was detected in 14 (25.4%) aborted fetuses. BHV-1 was isolated in MDBK cells from the tissue of 5 (9.1%) fetuses. The specificity of positive results was evaluated by Restriction Fragment Length Polymorphism (RFLP) with Bgl I restriction of DNA amplified by SN-PCR, and by virus-neutralization and immunofluorescence with rabbit anti-BHV-1 polyclonal antibodies for virus isolated in cell culture. The results of this work demonstrate the importance of using other diagnostic techniques, like SN-PCR, for BHV-1 detection in organ fragments from aborted fetuses and the high frequency of this virus in reproductive failures in Brazilian cattle herds.
采用半巢式聚合酶链反应（SN-PCR）以及MDBK细胞培养病毒分离法，对从巴西巴拉那州北部有繁殖问题病史的肉牛和奶牛群中采集的55份流产胎儿的器官碎片中的牛疱疹病毒1型（BHV-1）进行了调查。在14份（25.4%）流产胎儿中检测到了BHV-1糖蛋白D基因的425 bp扩增子。从5份（9.1%）胎儿的组织中在MDBK细胞中分离出了BHV-1。通过对SN-PCR扩增的DNA进行Bgl I酶切的限制性片段长度多态性（RFLP）分析，以及对细胞培养中分离出的病毒用兔抗BHV-1多克隆抗体进行病毒中和及免疫荧光分析，评估了阳性结果的特异性。这项工作的结果表明，使用其他诊断技术如SN-PCR来检测流产胎儿器官碎片中的BHV-1具有重要意义，以及该病毒在巴西牛群繁殖失败中出现的频率很高。
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