Performance evaluation of TaqMan™ Arbovirus Triplex Kit (ZIKV/DENV/CHIKV) for detection and differentiation of dengue and chikungunya viral RNA in serum samples of symptomatic patients

Abstract

Introduction: 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.

Methods: 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.

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.

Conclusion: 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.

Keywords: CDC DENV-1–4 rRT-PCR: CDC DENV-1–4 Real-Time RT-PCR assay; CDC Trioplex: Trioplex Real-time RT-PCR assay; CHIKV: Chikungunya virus; DENV: Dengue virus; TaqMan™ Arbovirus kit: TaqMan™ Arbovirus Triplex kit (ZIKV/DENV/CHIKV); ZIKV: Zika virus; rtRT-PCR: Real-time reverse transcriptase polymerase chain reaction.