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首页> 外文期刊>Journal of Clinical Microbiology >Direct Detection of Respiratory Syncytial Virus, Parainfluenza Virus, and Adenovirus in Clinical Respiratory Specimens by a Multiplex Reverse Transcription-PCR Assay
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Direct Detection of Respiratory Syncytial Virus, Parainfluenza Virus, and Adenovirus in Clinical Respiratory Specimens by a Multiplex Reverse Transcription-PCR Assay

机译：通过多重逆转录-PCR法直接检测临床呼吸道标本中的呼吸道合胞病毒，副流感病毒和腺病毒

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摘要

Diagnosis of respiratory virus infections currently involves detection by isolation or antigen detection, which usually identifies only a single suspected agent. To permit identification of more than one respiratory virus in clinical specimens, a rapid detection method involving a single-step, multiplex reverse transcription-PCR (RT-PCR) assay was developed. The assay included five primer sets that amplified the RNA of respiratory syncytial virus subtypes A and B, parainfluenza virus types 1, 2, and 3, and adenovirus types 1 to 7. Initially the assay was tested on tissue culture-grown virus and was found to be specific for all 12 prototype viruses tested, with no interassay cross amplification or amplification of other respiratory viruses. Assay sensitivity allowed a detection range of 0.2 50% tissue culture infectious dose (TCID50) for adenovirus to 250 TCID50 for parainfluenza virus type 1. The multiplex RT-PCR assay was also able to directly detect viruses in respiratory specimens, with virus being detected in 41 of 112 samples as compared to 34 of 112 samples detected by direct immunofluorescence or antigen detection following specimen culture. This suggests that the multiplex RT-PCR assay can be used as a rapid and sensitive diagnostic method for major respiratory viruses.
机译：呼吸道病毒感染的诊断目前涉及通过分离或抗原检测进行检测，通常仅可识别一种可疑病原体。为了能够在临床标本中鉴定出一种以上的呼吸道病毒，开发了一种快速检测方法，该方法涉及一步，多重逆转录PCR（RT-PCR）分析。该测定包括五种引物组，可扩增呼吸道合胞病毒A和B亚型，副流感病毒1、2和3型以及1至7型腺病毒的RNA。最初，对组织培养生长的病毒进行了测试，结果发现对所有测试的12种原型病毒都具有特异性，没有测定间交叉扩增或其他呼吸道病毒的扩增。检测灵敏度使腺病毒的检测范围为0.2 50％组织培养感染剂量（TCID 50 ）到1型副流感病毒的250 TCID 50 。多重RT-PCR检测该技术还能够直接检测呼吸道标本中的病毒，与在标本培养后通过直接免疫荧光或抗原检测法检测到的112个样品中的34个相比，在112个样品中的41个检测到病毒。这表明，多重RT-PCR检测可用作主要呼吸道病毒的快速灵敏诊断方法。

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