Validation of a Rapid Rabies Diagnostic Tool for Field Surveillance in Developing Countries
Validation of a Rapid Rabies Diagnostic Tool for Field Surveillance in Developing Countries
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DOI：
10.1371/journal.pntd.0005010
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发表时间：
2016-10-01
影响因子：
3.8
通讯作者：
Dacheux, Laurent
中科院分区：
文献类型：
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作者：
Lechenne, Monique;Naissengar, Kemdongarti;Dacheux, Laurent
狂犬病被忽视的一个根本原因是低收入国家缺乏足够的诊断测试。一种快速、性能友好和低成本的方法来检测脑样本中的狂犬病病毒（RABV），将有助于监测，从而有助于准确的数据报告，方法学/主要发现我们评价了快速免疫诊断试验（RIDT）与标准荧光抗体试验（FAT）的比较。并通过真实的时间逆转录聚合酶链反应（RT-qPCR）证实了病毒RNA的检测。我们的分析是一个多中心的方法，以验证RIDT的性能在现场实验室（恩贾梅纳，乍得）和国际参考实验室（巴斯德研究所，巴黎，法国）。在现场实验室中，对48份犬样本进行了检测，在参考实验室环境中，共检测了73份样本，代表了RABV在检测动物种属（13个不同种属）、分离株的地理来源（特别强调非洲）和不同系统发育分支方面的广泛多样性。在参考实验室条件下，与金标准FAT试验相比，特异性为93.3%，灵敏度为95.3%。在现场实验室条件下，RIDT产生了更高的可靠性比FAT测试，特别是对新鲜和分解的样品。随后直接从测试滤纸中提取病毒RNA，并进一步成功地用于测序和genotyping.Conclusion/SignificanceThe RIDT在用户友好性和结果可靠性方面均表现出优异的性能。此外，检测卡可用作将病毒RNA运送至参考实验室的载体，以进一步进行诊断的实验室确认和使用核苷酸测序进行流行病学调查。因此，在偏远地区令人满意的使用潜力非常高，以提高全球对狂犬病流行病学的认识。然而，我们建议在推广和更广泛使用之前对该方案进行一些修改，并进行仔细的进一步验证。
BackgroundOne root cause of the neglect of rabies is the lack of adequate diagnostic tests in the context of low income countries. A rapid, performance friendly and low cost method to detect rabies virus (RABV) in brain samples will contribute positively to surveillance and consequently to accurate data reporting, which is presently missing in the majority of rabies endemic countries.Methodology/Principal findingsWe evaluated a rapid immunodiagnostic test (RIDT) in comparison with the standard fluorescent antibody test (FAT) and confirmed the detection of the viral RNA by real time reverse transcription polymerase chain reaction (RT-qPCR). Our analysis is a multicentre approach to validate the performance of the RIDT in both a field laboratory (N'Djamena, Chad) and an international reference laboratory (Institut Pasteur, Paris, France). In the field laboratory, 48 samples from dogs were tested and in the reference laboratory setting, a total of 73 samples was tested, representing a wide diversity of RABV in terms of animal species tested (13 different species), geographical origin of isolates with special emphasis on Africa, and different phylogenetic clades. Under reference laboratory conditions, specificity was 93.3% and sensitivity was 95.3% compared to the gold standard FAT test. Under field laboratory conditions, the RIDT yielded a higher reliability than the FAT test particularly on fresh and decomposed samples. Viral RNA was later extracted directly from the test filter paper and further used successfully for sequencing and genotyping.Conclusion/SignificanceThe RIDT shows excellent performance qualities both in regard to user friendliness and reliability of the result. In addition, the test cassettes can be used as a vehicle to ship viral RNA to reference laboratories for further laboratory confirmation of the diagnosis and for epidemiological investigations using nucleotide sequencing. The potential for satisfactory use in remote locations is therefore very high to improve the global knowledge of rabies epidemiology. However, we suggest some changes to the protocol, as well as careful further validation, before promotion and wider use.