在基孔肯雅病毒、寨卡病毒和登革热病毒流行地区对多重虫媒病毒PCR检测试验的评估：按照临床实验室改进修正案（CLIA）标准对试剂盒性能特征的评估。
Assessment of a multiplex arbovirus PCR Detection Test in an area endemic for Chikungunya, Zika, and Dengue viruses: An evaluation of kit performance characteristics in line with Clinical Laboratory Improvement Amendments (CLIA) Standards.
作者信息
Kingwara Leonard, Odeny Lazarus, Onwonga Vera, Madada Rukia, Mbeneka Marygorett, Okonji Jully, Mwende Victoria, Rombo Charles, Akasa Shalyn, Ndia Millicent, Ouma Fredrick, Ayagah Isabella, Roba Abdi, Kimenye Kamene
机构信息
Kenya National Public Health Institute, Kenya.
Amref Health Africa, Africa.
出版信息
PLoS One. 2025 Jun 24;20(6):e0309626. doi: 10.1371/journal.pone.0309626. eCollection 2025.
INTRODUCTION
Several multiplex and singleplex PCR systems exist for the detection of Dengue(DENV), Zika(ZIKV), and Chikungunya(CHIKV) viruses; however, their performance characteristics in East Africa remain unassessed. In this study, we investigated the TaqMan® Arbovirus Triplex Kit, which allows for the simultaneous identification of CHIKV, DENV, and ZIKV viruses in serum samples and singleplex assays, including the RealStar® Chikungunya RT-PCR Kit 2.0, RealStar® Dengue RT-PCR Kit 3.0, and RealStar® Zika Virus RT-PCR Kit 1.0.
METHODS
We used Clinical Laboratory Improvement validation methods and residual specimens sourced from DENV, CHIKV, and ZIKV outbreaks, from Kenya, Brazil, and the Democratic Republic of Congo(DRC) in March 2024. Quality control was ensured through Quality Control for Medical diagnostics (QCMD) commercial positive panels. Gold standard results were established through consensus from multiple kits, including the TaqMan® Arbovirus Triplex Kit, Illumina viral surveillance panel (Illumina VSP), and Altona singleplex kit. Data were analyzed using R to determine specificity, sensitivity, negative predictive value (NPV), positive predictive value (PPV), and diagnostic odds ratio, with probit analysis employed to evaluate the limit of detection.
RESULTS
The TaqMan® Arbovirus Kit showed sensitivity exceeding 95% for all three virus targets: 97.98% for DENV, and 100% for both CHIKV and ZIKV. Simillarly, the RealStar® RT-PCR Kits exhibited sensitivities: 93.94% for CHIKV, 90.91% for DENV, and 100% for ZIKV. All kits demonstrated specificity above 99%, with a positive predictive value (PPV) over 99% and a negative predictive value (NPV) exceeding 97%. Probit analysis revealed that the TaqMan® Arbovirus Kit also had a lower detection limit compared to the RealStar kit for all tested viruses.
CONCLUSION
The TaqMan® Arbovirus Kit provides multiplex capabilities and performance similar to the RealStar® CHIKV, DENV, and ZIKV RT-PCR Kits. Consequently, these kits can be used interchangeably for detecting DENV, CHIKV, and ZIKV viruses in outbreak situations.
引言
现已有多种用于检测登革热病毒（DENV）、寨卡病毒（ZIKV）和基孔肯雅病毒（CHIKV）的多重和单重PCR系统；然而，它们在东非的性能特征仍未得到评估。在本研究中，我们调查了TaqMan®虫媒病毒三重检测试剂盒，该试剂盒可同时鉴定血清样本中的CHIKV、DENV和ZIKV病毒以及单重检测试剂盒，包括RealStar®基孔肯雅病毒RT-PCR试剂盒2.0、RealStar®登革热病毒RT-PCR试剂盒3.0和RealStar®寨卡病毒RT-PCR试剂盒1.0。
方法
我们采用了临床实验室改进验证方法，并使用了2024年3月从肯尼亚、巴西和刚果民主共和国（DRC）的DENV、CHIKV和ZIKV疫情中获取的剩余样本。通过医学诊断质量控制（QCMD）商业阳性对照确保质量控制。通过包括TaqMan®虫媒病毒三重检测试剂盒、Illumina病毒监测面板（Illumina VSP）和Altona单重检测试剂盒在内的多种试剂盒达成共识来确定金标准结果。使用R软件分析数据以确定特异性、敏感性、阴性预测值（NPV）、阳性预测值（PPV）和诊断比值比，并采用概率分析来评估检测限。
结果
TaqMan®虫媒病毒试剂盒对所有三种病毒靶点的敏感性均超过95%：对DENV为97.98%，对CHIKV和ZIKV均为100%。同样，RealStar® RT-PCR试剂盒的敏感性分别为：CHIKV为93.94%，DENV为90.91%，ZIKV为100%。所有试剂盒的特异性均高于99%，阳性预测值（PPV）超过99%，阴性预测值（NPV）超过97%。概率分析表明，对于所有测试病毒，TaqMan®虫媒病毒试剂盒的检测限也低于RealStar试剂盒。
结论
TaqMan®虫媒病毒试剂盒提供了与RealStar® CHIKV、DENV和ZIKV RT-PCR试剂盒相似的多重检测能力和性能。因此，在疫情情况下，这些试剂盒可互换使用以检测DENV、CHIKV和ZIKV病毒。
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