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首页> 外文期刊>Journal of Clinical Microbiology >Evaluation of a Nested Reverse Transcription-PCR Assay Based on the Nucleoprotein Gene for Diagnosis of Spontaneous and Experimental Bovine Respiratory Syncytial Virus Infections
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Evaluation of a Nested Reverse Transcription-PCR Assay Based on the Nucleoprotein Gene for Diagnosis of Spontaneous and Experimental Bovine Respiratory Syncytial Virus Infections

机译：基于核蛋白基因的巢式逆转录-PCR分析的评估，用于诊断自发性和实验性牛呼吸道合胞病毒感染

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摘要

The first nested reverse transcription (RT)-PCR based on the nucleoprotein gene (n RT-PCR-N) of the bovine respiratory syncytial virus (BRSV) has been developed and optimized for the detection of BRSV in bronchoalveolar lavage fluid cells of calves. This test is characterized by a low threshold of detection (0.17 PFU/ml), which is 506 times lower than that obtained by an enzyme immunosorbent assay (EIA) test (RSV TESTPACK ABBOTT). During an experimental infection of 17 immunocompetent calves less than 3 months old, BRSV RNA could be detected up to 13 days after the onset of symptoms whereas isolation in cell culture was possible only up to 5 days. Compiling results obtained by conventional techniques (serology, antigen detection, and culture isolation) for 132 field samples collected from calves with acute respiratory signs revealed that n RT-PCR-N showed the highest diagnostic sensitivity and very good specificity. This n RT-PCR-N with its long period of detection during BRSV infection thus provides a valuable tool for diagnostic and epidemiological purposes.
机译：基于牛呼吸道合胞病毒（BRSV）的核蛋白基因（n RT-PCR-N）的第一个嵌套逆转录（RT）-PCR已经开发并优化，用于犊牛支气管肺泡灌洗液细胞中的BRSV检测。此测试的特点是检测阈值低（0.17 PFU / ml），比酶免疫吸附测定（EIA）测试（RSV TESTPACK ABBOTT）的检测阈值低506倍。在实验性感染不足3个月大的17只免疫活性小牛的过程中，BRSV RNA可以在症状发作后的13天之内被检测到，而在细胞培养中分离到5天才可以。通过常规技术（血清学，抗原检测和培养物分离）对从具有急性呼吸道症状的小牛收集的132个田间样品收集的结果表明，n RT-PCR-N显示出最高的诊断灵敏度和非常好的特异性。因此，这种nRT-PCR-N在BRSV感染期间具有很长的检测时间，因此为诊断和流行病学目的提供了有价值的工具。

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