Starick E
Federal Research Centre for Virus Diseases of Animals, Friedrich-Loeffler-Institute, Insel Riems, Germany.
Acta Virol. 1998 Nov;42(5):333-9.
A reverse transcription-polymerase chain reaction (RT-PCR) assay using four different primer pairs for the detection of equine arteritis virus (EAV) RNA in semen and tissue samples was evaluated. A fragment encoding the leader sequence of the EAV genome was most successfully amplified. The specificity and sensitivity of RT-PCR was assessed by virus isolation in cell culture, restriction analysis, dot blot hybridisation and nested PCR. To this end, 23 semen samples from seropositive stallions and 11 tissue samples from 4 aborted foals were tested. Compared to the virus isolation test in cell culture, the sensitivity of the molecular methods proved to be higher. In the RT-PCR, dot blot hybridisation and nested PCR tests, semen samples from 11 stallions and tissue samples from all 4 foals were found positive, while the virus could be isolated in cell culture from only 4 semen samples and tissue samples from 1 aborted foal. The sensitivity of the dot blot hybridisation test was superior to that of the RT-PCR test. The nested PCR test proved to be the most sensitive one, because 3 semen samples were recognised as positive by this method only. Considering the sensitivity, and rapidity and reliability, RT-PCR followed by dot blot hybridisation or nested PCR represents the best method for diagnosis of EAV and should be included in the official diagnostic regimes.
评估了一种逆转录-聚合酶链反应（RT-PCR）检测方法，该方法使用四对不同引物对检测精液和组织样本中的马动脉炎病毒（EAV）RNA。编码EAV基因组前导序列的片段扩增最为成功。通过细胞培养中的病毒分离、限制性分析、斑点杂交和巢式PCR评估RT-PCR的特异性和敏感性。为此，检测了23份来自血清学阳性种马的精液样本和11份来自4匹流产驹的组织样本。与细胞培养中的病毒分离试验相比，分子方法的敏感性更高。在RT-PCR、斑点杂交和巢式PCR试验中，发现11匹种马的精液样本和所有4匹驹的组织样本呈阳性，而在细胞培养中仅能从4份精液样本和1匹流产驹的组织样本中分离出病毒。斑点杂交试验的敏感性优于RT-PCR试验。巢式PCR试验被证明是最敏感的，因为只有该方法识别出3份精液样本呈阳性。考虑到敏感性、快速性和可靠性，RT-PCR随后进行斑点杂交或巢式PCR是诊断EAV的最佳方法，应纳入官方诊断方案。
