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Searching for the best real-time RT-PCRs to detect Zika virus infections: the importance of comparing several protocols

机译:寻找检测寨卡病毒感染的最佳实时RT-PCR:比较几种方案的重要性

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摘要

Clinical manifestations of Zika, dengue, and chikungunya virus infections are very similar, making it difficult to reach a diagnosis based only on clinical grounds. In addition, there is an intense cross-reactivity between antibodies directed to Zika virus and other flaviviruses, and an accurate Zika diagnosis is best achieved by real-time RT-PCR. However, some real-time RT-PCR show better performance than others. To reach the best possible Zika diagnosis, the analytic sensitivity of some probe-based real-time RT-PCR amplifying Zika virus RNA was evaluated in spiked and clinical samples. We evaluated primers and probes to detect Zika virus, which had been published before, and tested sensitivity using serum spiked and patient samples by real-time RT-PCR. When tested against spiked samples, the previously described primers showed different sensitivity, with very similar results when samples from patients (serum and urine) were analyzed. Real-time RT-PCR designed to amplify Zika virus NS1 showed the best analytical sensitivity for all samples.
机译:寨卡病毒、登革热病毒和基孔肯雅病毒感染的临床表现非常相似,因此很难仅根据临床依据做出诊断。此外,针对寨卡病毒和其他黄病毒的抗体之间存在强烈的交叉反应性,实时RT-PCR最好实现准确的寨卡诊断。然而,一些实时荧光定量RT-PCR显示出比其他的更好的性能。为了达到最佳的寨卡诊断,在加标样本和临床样本中评估了一些基于探针的实时RT-PCR扩增寨卡病毒RNA的分析灵敏度。我们评估了之前发表的用于检测寨卡病毒的引物和探针,并通过实时RT-PCR测试了血清加标样本和患者样本的灵敏度。当对加标样品进行测试时,先前描述的引物显示出不同的灵敏度,当分析来自患者的样品(血清和尿液)时,结果非常相似。旨在扩增寨卡病毒NS1的实时RT-PCR对所有样品都显示出最佳的分析灵敏度。

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