Laboratory of Clinical Virology, Department of Medical Microbiology, Academic Medical Centre, University of Amsterdam, PO-Box 22600, 1100 DD, Amsterdam, The Netherlands.
J Clin Virol. 2011 Jul;51(3):179-85. doi: 10.1016/j.jcv.2011.04.010. Epub 2011 May 14.
Multiplex real time PCR is increasingly used to diagnose respiratory viruses and has shown to be superior to traditional methods, like culture and antigen detection. However, comprehensive data on sensitivity, specificity and performance of the multiplex PCR compared to the single target PCR's is limited for most published respiratory multiplex real time PCR assays.
Development and extensive analysis of an internally controlled multiplex real time rt-PCR for detection of respiratory viruses.
The assay was validated in comparison to single-target PCR's using plasmid targets and prospectively collected nasopharyngeal aspirates.
Using plasmid targets the multiplex format was found to be as least as sensitive and specific as the single-target PCR and no competition was observed when different targets were present at different amounts in one tube. Clinical validation showed high concordance for all viruses tested except for samples with low levels of enterovirus.
This multiplex showed excellent specificities for all 14 respiratory viruses and sensitivity was high except for clinical samples with low levels of enterovirus.
多重实时 PCR 越来越多地用于诊断呼吸道病毒，并且已经被证明优于传统方法，如培养和抗原检测。然而，对于大多数已发表的呼吸道多重实时 PCR 检测，与单靶标 PCR 相比，关于多重 PCR 的灵敏度、特异性和性能的综合数据是有限的。
开发和广泛分析一种内部对照的多重实时逆转录 PCR 检测呼吸道病毒的方法。
该检测方法通过质粒靶标和前瞻性收集的鼻咽抽吸物与单靶标 PCR 进行了比较。
使用质粒靶标，发现多重格式与单靶标 PCR 一样敏感和特异，并且当不同靶标以不同数量存在于一个管中时，没有观察到竞争。临床验证显示，除了低水平肠道病毒的样本外，所有测试的病毒都有很高的一致性。
这种多重方法对所有 14 种呼吸道病毒显示出极好的特异性，除了低水平肠道病毒的临床样本外，灵敏度也很高。
