Panmei Kakhangchung, Syed Abdul Hakeem, Okafor Obiageli, Mammen Shoba, Abraham Asha Mary
Dept. of Clinical Virology, Christian Medical College, Vellore, Tamil Nadu, India.
Thermo Fisher Scientific, South San Francisco, USA.
J Virol Methods. 2025 Feb;332:115072. doi: 10.1016/j.jviromet.2024.115072. Epub 2024 Nov 20.
Global outbreaks of mosquito-transmitted arbovirus infections, such as dengue (DENV) and chikungunya (CHIKV), are increasing. Differentiating these infections is challenging due to non-specific symptoms and serology limitations. PCR-based approaches offer higher sensitivity and specificity. This study evaluated the performance of TaqMan™ Arbovirus Triplex Kit (ZIKV/DENV/CHIKV) (TaqMan™ Kit) to detect DENV and CHIKV in clinical samples from patients in south India.
In total, 280 serum samples with 90 DENV-positive, 90 CHIKV-positive, and 100 negative samples were tested with TaqMan™ Kit and CDC Trioplex Real-Time RT-PCR assay. No Zika virus was detected. The sensitivity and specificity of viral RNA detection were determined, and discordant results were resolved using comparator PCRs, dengue NS1 antigen detection, virus-specific antibody results, or previously de-identified in-house PCR results.
TaqMan™ Kit showed 100 % agreement with the comparator for DENV detection in 92 positive samples. Among 188 samples negative for DENV by the comparator, 30 showed positive results with the TaqMan™ kit, and 23 of those were confirmed as true positives. The resulting sensitivity and specificity for DENV detection were 100 % and 95.1 %, respectively. For CHIKV, 77 positive and 195 negative results were concordant. Eight samples showed discordant results, but upon resolution testing, sensitivity and specificity for CHIKV were 93.9 % and 100.0 %, respectively.
TaqMan™ Arbovirus Triplex Kit demonstrated high sensitivity and specificity (>93 %) for detecting circulating DENV and CHIKV strains. Multiplex PCR testing can improve case detection, surveillance, and public health responses while optimizing laboratory resources for outbreak control.
登革热(DENV)和基孔肯雅热(CHIKV)等蚊媒传播虫媒病毒感染的全球暴发正在增加。由于症状不具特异性以及血清学存在局限性,区分这些感染具有挑战性。基于聚合酶链反应(PCR)的方法具有更高的灵敏度和特异性。本研究评估了TaqMan™虫媒病毒三重检测试剂盒(寨卡病毒/登革热病毒/基孔肯雅热病毒)(TaqMan™试剂盒)检测印度南部患者临床样本中登革热病毒和基孔肯雅热病毒的性能。
总共280份血清样本,其中90份登革热病毒阳性、90份基孔肯雅热病毒阳性和100份阴性样本,使用TaqMan™试剂盒和美国疾病控制与预防中心(CDC)三重实时逆转录聚合酶链反应(RT-PCR)检测法进行检测。未检测到寨卡病毒。确定了病毒RNA检测的灵敏度和特异性,并使用比较PCR、登革热非结构蛋白1(NS1)抗原检测、病毒特异性抗体结果或先前未识别的内部PCR结果解决不一致的结果。
在92份登革热病毒阳性样本中,TaqMan™试剂盒与比较检测法对登革热病毒检测的一致性为100%。在比较检测法检测为登革热病毒阴性的188份样本中,30份用TaqMan™试剂盒检测呈阳性,其中23份被确认为真阳性。登革热病毒检测的灵敏度和特异性分别为100%和95.1%。对于基孔肯雅热病毒,77份阳性和195份阴性结果一致。8份样本结果不一致,但经解决检测后,基孔肯雅热病毒检测的灵敏度和特异性分别为93.9%和100.0%。
TaqMan™虫媒病毒三重检测试剂盒在检测循环中的登革热病毒和基孔肯雅热病毒毒株方面表现出高灵敏度和特异性(>93%)。多重PCR检测可以改善病例检测、监测和公共卫生应对,同时优化用于疫情控制的实验室资源。