利用重组酶聚合酶扩增分析法对急性期尿液样本中的寨卡病毒进行快速分子检测。
Rapid Molecular Detection of Zika Virus in Acute-Phase Urine Samples Using the Recombinase Polymerase Amplification Assay.
作者信息
Abd El Wahed Ahmed, Sanabani Sabri Saeed, Faye Oumar, Pessôa Rodrigo, Patriota João Veras, Giorgi Ricardo Rodrigues, Patel Pranav, Böhlken-Fascher Susanne, Landt Olfert, Niedrig Matthias, Zanotto Paolo Marinho de Andrade, Czerny Claus Peter, Sall Amadou A, Weidmann Manfred
机构信息
Division of Microbiology and Animal Hygiene, Institute of Veterinary Medicine, Department of Animal Sciences, Georg-August-University, Goettingen, Lower Saxony, Germany.
Hospital das Clínicas, School of Medicine, University of São Paulo, São Paulo, Brazil.
出版信息
PLoS Curr. 2017 Jan 25;9:ecurrents.outbreaks.a7f1db2c7d66c3fc0ea0a774305d319e. doi: 10.1371/currents.outbreaks.a7f1db2c7d66c3fc0ea0a774305d319e.
BACKGROUND
Currently the detection of Zika virus (ZIKV) in patient samples is done by real-time RT-PCR. Samples collected from rural area are sent to highly equipped laboratories for screening. A rapid point-of-care test is needed to detect the virus, especially at low resource settings.
METHODOLOGY/PRINCIPAL FINDINGS: In this report, we describe the development of a reverse transcription isothermal recombinase polymerase amplification (RT-RPA) assay for the identification of ZIKV. RT-RPA assay was portable, sensitive (21 RNA molecules), and rapid (3-15 minutes). No cross-reactivity was detected to other flaviviruses, alphaviruses and arboviruses. Compared to real-time RT-PCR, the diagnostic sensitivity was 92%, while the specificity was 100%.
CONCLUSIONS/SIGNIFICANCE: The developed assay is a promising platform for rapid point of need detection of ZIKV in low resource settings and elsewhere (e.g. during mass gathering).
背景
目前，患者样本中寨卡病毒（ZIKV）的检测通过实时逆转录聚合酶链反应（RT-PCR）进行。从农村地区采集的样本被送往设备精良的实验室进行筛查。需要一种快速的即时检测方法来检测该病毒，尤其是在资源匮乏的地区。
方法/主要发现：在本报告中，我们描述了一种用于鉴定寨卡病毒的逆转录等温重组酶聚合酶扩增（RT-RPA）检测方法的开发。RT-RPA检测方法便于携带、灵敏（可检测21个RNA分子）且快速（3 - 15分钟）。未检测到与其他黄病毒、甲病毒和虫媒病毒的交叉反应。与实时RT-PCR相比，诊断敏感性为92%，特异性为100%。
结论/意义：所开发的检测方法是在资源匮乏地区及其他地方（如在大规模集会期间）对寨卡病毒进行快速即时检测的一个有前景的平台。
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