Department of Health Sciences, University of Genoa, 16132 Genoa, Italy.
Hygiene Unit, San Martino Policlinico Hospital, IRCCS for Oncology and Neurosciences, 16132 Genoa, Italy.
Medicina (Kaunas). 2024 Sep 23;60(9):1557. doi: 10.3390/medicina60091557.
: The steady spread of dengue virus (DENV) poses a profound public health threat worldwide. Reverse transcription real-time polymerase chain reaction (RT2-PCR) has been increasingly recognized as a reference method for the diagnosis of acute dengue infection. The goal of this study was to assess the diagnostic accuracy of five different RT2-PCR kits for the detection of DENV in a historically processed set of sera samples. : In this retrospective study, 25 sera samples from routinely processed unique adult patients with a known DENV status (previously tested in both molecular and serological assays) were tested in parallel using four conventional (RealStar Dengue PCR Kit 3.0, Clonit'ngo Zika, Dengue & Chikungunya, BioPerfectus Zika Virus/Dengue Virus/Chikungunya Virus Real Time PCR Kit and Novaplex Tropical fever virus) and one sample-to-result (STANDARD M10 Arbovirus Panel) RT2-PCR assays. Additionally, an end-point dilution analysis was conducted in quintuplicate on six serial dilutions of an RNA preparation obtained from a culture-grown DENV serotype 1 strain for a total of 150 tests. : The overall accuracy of the evaluated tests ranged from 84% to 100%. In particular, the sensitivity of three conventional RT2-PCR assays (RealStar, Clonit'ngo and Novaplex) was 100% (95% CI: 79.6-100%), while it was lower (73.3%; 95% CI: 48.1-89.1%) for the BioPerfectus kit. The sample-to-result STANDARD M10 panel performed comparatively well, showing a sensitivity of 92.9% (95% CI: 68.5-98.7%). No false positive results were registered in any assay. The end-point dilution analysis suggested that the RealStar kit had the lowest limit of detection. : Available RT2-PCR kits for the detection of DENV are highly specific and generally sensitive and, therefore, their implementation in diagnostic pathways is advisable.
登革热病毒(DENV)的稳定传播对全球公共卫生构成了严重威胁。逆转录实时聚合酶链反应(RT2-PCR)已越来越被认为是诊断急性登革热感染的参考方法。本研究旨在评估五种不同 RT2-PCR 试剂盒在一组历史处理血清样本中检测 DENV 的诊断准确性。
在这项回顾性研究中,使用四种常规 RT2-PCR 试剂盒(RealStar Dengue PCR Kit 3.0、Clonit'ngo Zika、Dengue & Chikungunya、BioPerfectus Zika Virus/Dengue Virus/Chikungunya Virus Real Time PCR Kit)和一种样本到结果(STANDARD M10 Arbovirus Panel)对 25 份来自常规处理的具有已知 DENV 状态的独特成年患者的血清样本进行了平行检测。此前已在分子和血清学检测中进行了检测)。此外,还对从培养的 DENV 血清型 1 株获得的 RNA 制剂的六个连续稀释度进行了终点稀释分析,共进行了 150 次测试。
评估测试的整体准确性范围为 84%至 100%。特别是,三种常规 RT2-PCR 检测(RealStar、Clonit'ngo 和 Novaplex)的灵敏度为 100%(95%CI:79.6-100%),而 BioPerfectus 试剂盒的灵敏度较低(73.3%;95%CI:48.1-89.1%)。样本到结果的 STANDARD M10 面板表现相对较好,灵敏度为 92.9%(95%CI:68.5-98.7%)。任何检测均未出现假阳性结果。终点稀释分析表明,RealStar 试剂盒的检测下限最低。
用于检测 DENV 的可用 RT2-PCR 试剂盒具有高度特异性,通常具有较高的灵敏度,因此建议将其纳入诊断途径。