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首页> 外文期刊>The American Journal of Tropical Medicine and Hygiene >A Direct from Blood/Plasma Reverse Transcription-Polymerase Chain Reaction for Dengue Virus Detection in Point-of-Care Settings
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A Direct from Blood/Plasma Reverse Transcription-Polymerase Chain Reaction for Dengue Virus Detection in Point-of-Care Settings

机译：一种直接来自血液/等离子体逆转录 - 聚合酶链反应，用于登记病毒检测在护理点设置

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摘要

Infection with dengue virus (DENV) is widespread across tropical regions and can result in severe disease. Early diagnosis is important both for patient management and to differentiate infections that present with similar symptoms, such as malaria, chikungunya, and Zika. Rapid diagnostic tests that are used presently for point-of-care detection of DENV antigens lack the sensitivity of molecular diagnostics that detect viral RNA. However, no molecular diagnostic test for DENV is available for use in field settings. In this study, we developed and validated a reverse transcription-polymerase chain reaction (RT-PCR) for the detection of DENV adapted for use in field settings. Reverse transcription-polymerase chain reaction was performed directly from plasma samples without RNAextraction. The assay detected all four serotypes of DENV spiked into blood or plasma. Our RT-PCR does not cross-react with pathogens that cause symptoms that overlap with dengue infection. The test performed equally well in a conventional laboratory qPCR instrument and a small, low-cost portable instrument that can be used in a field setting. The lower limit of detection for the assay was 1 x 10(4) genome copy equivalents/mL in blood. Finally, we validated our test using 126 archived patient samples. The sensitivity of our RT-PCR was 76.7% (95% CI: 65.8-87.9%) on the conventional instrument, and 78.3% (95% CI: 65.8-87.9%) on the field instrument, when compared with the RealStar Dengue RT-PCR Kit 2.0. The molecular test described here is user-friendly, low-cost, and can be used in regions with limited laboratory capabilities.
机译：用登革热病毒（DENV）感染在热带地区普遍存在，可能导致严重疾病。早期诊断对于患者管理是重要的，并且对患有类似症状的感染，例如疟疾，Chikungunya和Zika。目前用于DENV抗原的护理点检测使用的快速诊断测试缺乏检测病毒RNA的分子诊断的敏感性。但是，DENV没有分子诊断测试可用于现场设置。在该研究中，我们开发并验证了逆转录聚合酶链反应（RT-PCR），用于检测适用于现场设置的DENV。逆转录 - 聚合酶链反应直接来自血浆样品，无需RNAExtraction。该测定检测到DENV的所有四种血清型掺入血液或血浆中。我们的RT-PCR不会与病原体交叉反应，导致与登革热感染重叠的症状。该测试在传统的实验室QPCR仪器和可用于场设置中使用的小型低成本便携式仪器进行。测定的检测下限为1×10（4）个基因组复制当量/ ml血液中。最后，我们使用126份患者样本验证了我们的测试。与Realstar Dengue RT相比，我们的RT-PCR的敏感性为常规仪器上的76.7％（95％CI：65.8-87.9％），域内仪器78.3％（95％CI：65.8-87.9％） -PCR套件2.0。这里描述的分子测试是用户友好的，低成本，并且可以用于具有有限的实验室能力的区域。

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