Létant Sonia E, Ortiz Josue I, Bentley Tammero Lance F, Birch James M, Derlet Robert W, Cohen Stuart, Manning Dannelle, McBride Mary T
Lawrence Livermore National Laboratory, L-231, Livermore, CA 94550, USA.
J Clin Microbiol. 2007 Nov;45(11):3498-505. doi: 10.1128/JCM.01712-07. Epub 2007 Sep 12.
We have developed a nucleic acid-based assay that is rapid, sensitive, and specific and can be used for the simultaneous detection of five common human respiratory pathogens, including influenza virus A, influenza virus B, parainfluenza virus types 1 and 3, respiratory syncytial virus (RSV), and adenovirus groups B, C, and E. Typically, diagnosis on an unextracted clinical sample can be provided in less than 3 h, including sample collection, preparation, and processing, as well as data analysis. Such a multiplexed panel would enable rapid broad-spectrum pathogen testing on nasal swabs and therefore allow implementation of infection control measures and the timely administration of antiviral therapies. We present here a summary of the assay performance in terms of sensitivity and specificity. The limits of detection are provided for each targeted respiratory pathogen, and result comparisons were performed on clinical samples, our goal being to compare the sensitivity and specificity of the multiplexed assay to the combination of immunofluorescence and shell vial culture currently implemented at the University of California-Davis Medical Center hospital. Overall, the use of the multiplexed reverse transcription-PCR assay reduced the rate of false-negative results by 4% and reduced the rate of false-positive results by up to 10%. The assay correctly identified 99.3% of the clinical negatives and 97% of the adenovirus, 95% of the RSV, 92% of the influenza virus B, and 77% of the influenza virus A samples without any extraction performed on the clinical samples. The data also showed that extraction will be needed for parainfluenza virus, which was only identified correctly 24% of the time on unextracted samples.
我们开发了一种基于核酸的检测方法，该方法快速、灵敏且特异，可用于同时检测五种常见的人类呼吸道病原体，包括甲型流感病毒、乙型流感病毒、1型和3型副流感病毒、呼吸道合胞病毒（RSV）以及B、C和E组腺病毒。通常情况下，在不到3小时内就能对未提取的临床样本提供诊断结果，包括样本采集、制备、处理以及数据分析。这样一个多重检测面板能够对鼻拭子进行快速的广谱病原体检测，从而得以实施感染控制措施并及时给予抗病毒治疗。我们在此呈现该检测方法在灵敏度和特异性方面的性能总结。给出了每种目标呼吸道病原体的检测限，并对临床样本进行了结果比较，我们的目的是将多重检测方法的灵敏度和特异性与加利福尼亚大学戴维斯分校医学中心医院目前采用的免疫荧光和空斑培养法相结合的方法进行比较。总体而言，使用多重逆转录 - PCR检测方法可将假阴性结果率降低4%，并将假阳性结果率降低多达10%。在未对临床样本进行任何提取的情况下，该检测方法正确识别了99.3%的临床阴性样本、97%的腺病毒样本、95%的呼吸道合胞病毒样本、92%的乙型流感病毒样本以及77%的甲型流感病毒样本。数据还表明，对于副流感病毒，需要进行提取，因为在未提取的样本上，其仅在24%的时间内被正确识别。
