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干棉拭子或植绒呼吸道拭子作为一种使用实时核酸序列扩增技术进行呼吸道病毒分子检测的简单采集技术。

Dry cotton or flocked respiratory swabs as a simple collection technique for the molecular detection of respiratory viruses using real-time NASBA.

作者信息

Moore Catherine, Corden Sally, Sinha Jaisi, Jones Rachel

机构信息

Wales Specialist Virology Centre, University Hospital of Wales, Heath Park, Cardiff, CF14 4XW, United Kingdom.

出版信息

J Virol Methods. 2008 Nov;153(2):84-9. doi: 10.1016/j.jviromet.2008.08.001. Epub 2008 Sep 11.

Abstract

This paper describes the molecular detection of influenza A, influenza B, respiratory syncytial virus and human metapneumovirus using real-time nucleic acid sequence based amplification (NASBA) from respiratory samples collected on simple dry cotton swabs, non-invasively and in the absence of transport medium. Viral RNA was detectable on dry cotton and flocked swabs for at least 2 weeks at room temperature and was readily extracted using magnetic silica extraction methods. Dry cotton respiratory swabs were matched with traditionally collected respiratory samples from the same patient, and results of traditional laboratory techniques and real-time NASBA were compared for all four viral targets. The results not only showed a significant increase in the detection rate of the viral targets over traditional laboratory methods of 46%, but also that dry swabs did not compromise their recovery. Over two subsequent winter seasons, 736 dry cotton respiratory swabs were collected from symptomatic patients and tested using real-time NASBA giving an overall detection rate for these respiratory virus targets of 38%. The simplicity of the method together with the increased detection rate observed in the study proves that transporting a dry respiratory swab to the laboratory for respiratory virus diagnosis using molecular methods is a suitable and robust alternative to traditional sample types.

摘要

本文描述了使用基于实时核酸序列扩增(NASBA)技术,对收集于简单干燥棉拭子上的呼吸道样本进行甲型流感病毒、乙型流感病毒、呼吸道合胞病毒和人偏肺病毒的分子检测。该检测无需运输培养基,非侵入性操作。室温下,干燥棉拭子和植绒拭子上的病毒RNA至少两周内均可检测到,且使用磁性硅胶提取方法可轻松提取病毒RNA。将干燥棉拭子与同一患者传统采集的呼吸道样本进行匹配,并比较了针对所有四种病毒靶点的传统实验室技术和实时NASBA的检测结果。结果不仅显示病毒靶点的检测率比传统实验室方法显著提高了46%,还表明干燥拭子不会影响病毒的回收率。在随后的两个冬季,从有症状的患者中收集了736份干燥棉拭子呼吸道样本,并使用实时NASBA进行检测,这些呼吸道病毒靶点的总体检测率为38%。该方法的简便性以及研究中观察到的检测率提高,证明了将干燥呼吸道拭子运送到实验室,使用分子方法进行呼吸道病毒诊断,是传统样本类型的一种合适且可靠的替代方法。

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