Diagnostics-in-a-Suitcase: Development of a portable and rapid assay for the detection of the emerging avian influenza A (H7N9) virus
Diagnostics-in-a-Suitcase: Development of a portable and rapid assay for the detection of the emerging avian influenza A (H7N9) virus
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DOI：
10.1016/j.jcv.2015.05.004
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发表时间：
2015-08-01
影响因子：
8.8
通讯作者：
Hufert, Frank T.
中科院分区：
文献类型：
--
作者：
Abd El Wahed, Ahmed;Weidmann, Manfred;Hufert, Frank T.
背景资料：在发展中国家，只有少数几个中心实验室才有诊断所需的设备，这些实验室不太容易获得，而且检测大量输入样本的能力有限。此外，样本的运输条件不充分，因此导致结果不可靠。目的：开发一种快速、廉价和简单的检测方法，可以移动的检测病毒。研究设计：手提箱实验室“手提箱诊断”（56 cm x 45.5 cm x 26.5 cm），包含进行逆转录重组酶聚合酶扩增（RT-PCR）所需的所有试剂和装置。RPA）测定。结果：H7和N9两种RT-RPA检测方法的灵敏度分别为10和100个RNA分子。在单一温度（42 ° C）下进行测定。H7N9 RT-RPA检测没有显示出对影响人类和/或鸟类的任何其他呼吸道病毒或人类或鸡基因组的交叉检测。所有试剂均在环境温度下使用、储存和运输，即不依赖冷链。此外，诊断在一个手提箱是由一个太阳能电池。结论：开发的检测协议和移动的设置进行良好。此外，它可以很容易地实现在机场，检疫站或农场进行诊断，以快速进行现场病毒核酸检测。(C)2015作者由爱思唯尔公司出版
Background: In developing countries, equipment necessary for diagnosis is only available in few central laboratories, which are less accessible and of limited capacity to test large numbers of incoming samples. Moreover, the transport conditions of samples are inadequate, therefore leading to unreliable results.Objectives: The development of a rapid, inexpensive, and simple test would allow mobile detection of viruses.Study design: A suitcase laboratory "Diagnostics-in-a-Suitcase" (56cm x 45.5 cm x 26.5 cm) containing all reagents and devices necessary for performing a reverse transcription recombinase polymerase amplification (RT-RPA) assay was developed. As an example, two RT-RPA assays were established for the detection of hemagglutinin (H) and neuraminidase ( N) genes of the novel avian influenza (H7N9) virus.Results: The sensitivities of the H7 and the N9 RT-RPA assays were 10 and 100 RNA molecules, respectively. The assays were performed at a single temperature (42 degrees C). The results were obtained within 2-7 min. The H7N9 RT-RPA assays did not show a cross-detection either of any other respiratory viruses affecting humans and/or birds or of the human or chicken genomes. All reagents were used, stored, and transported at ambient temperature, that is, cold chain independent. In addition, the Diagnostics-in-a-Suitcase was operated by a solar-powered battery.Conclusions: The developed assay protocol and mobile setup performed well. Moreover, it can be easily implemented to perform diagnoses at airports, quarantine stations, or farms for rapid on-site viral nucleic acid detection. (C) 2015 The Authors. Published by Elsevier B.V.