Abstract
.Recent expansions of vector-borne diseases highlight the need for improved surveillance, especially in resource-poor settings. Dengue virus (DENV), chikungunya virus (CHIKV), and Zika virus (ZIKV) share the same vectors as well as similar clinical presentations, suggesting that combined surveillance would be useful. We hypothesized that blood spotted on dengue rapid diagnostic tests (RDTs) could be harnessed for sample collection in remote areas for subsequent detection of DENV, CHIKV, and ZIKV by reverse transcription real-time polymerase chain reaction (RT-qPCR). CHIKV and ZIKV dilutions were spotted on dengue RDTs (SD BIOLINE Dengue DUO, Standard Diagnostics, Gyeonggi-do, Republic of Korea), dried, and extracted. As reference, aliquots of each viral dilution were directly extracted. Using specific RT-qPCR tests, both viruses were successfully detected from RDT extracts. However, the limit of detection was slightly lower in comparison to direct extracts, two logfold for CHIKV and one logfold for ZIKV. For analysis of temperature stability, DENV dilutions were spotted on RDTs and stored for up to 2 months at −80°C, 4°C, or 35°C before testing. Storage of RDTs for 2 months at 35°C did not compromise detection of RNA by RT-qPCR; only minimal degradation was observed. This proof-of-principle study demonstrates the potential of using dengue RDTs for DENV/CHIKV/ZIKV combined surveillance in areas without access to laboratory facilities. Further investigations are needed for evaluation of tri-viral surveillance under field conditions using patient samples. Large-scale implementation of surveillance for these viruses is of crucial public health importance for the early detection of epidemics. This method also has important implications for improving understanding of the molecular epidemiology of the three viruses.
Highlights
Dengue virus (DENV) infection is hyperendemic in tropical and subtropical areas, with an estimated 390 million patients per year.[1,2] There is recent evidence to suggest alarming global expansion of two other mosquito-borne viruses, chikungunya (CHIKV) and Zika (ZIKV) viruses.[3,4,5] In Southeast Asia (SEA), urban outbreaks of chikungunya virus (CHIKV) have been well documented in the past decade, but epidemiology in rural areas is not well characterized.[6]
DENV RT-qPCR results from the rapid diagnostic tests (RDTs) loaded with the three DENV1 dilutions and stored at different temperatures are displayed in Table 2 and supplemental data (Supplemental Table 1)
For the “low-titer” dilution, DENV RNA could not be amplified for one of the three RDTs stored at 35°C for 1 week, and for the three RDTs stored at 35°C for 2 months
Summary
Dengue virus (DENV) infection is hyperendemic in tropical and subtropical areas, with an estimated 390 million patients per year.[1,2] There is recent evidence to suggest alarming global expansion of two other mosquito-borne viruses, chikungunya (CHIKV) and Zika (ZIKV) viruses.[3,4,5] In Southeast Asia (SEA), urban outbreaks of CHIKV have been well documented in the past decade, but epidemiology in rural areas is not well characterized.[6]. With only limited data available, ZIKV circulation is probably underestimated.[7] a few countries reported increasing number of patients in 2016–2017, 686 in Thailand, 493 in Singapore, 232 in Vietnam, and 57 patients in the Philippines.[8] The three arboviruses are transmitted by the same mosquito vectors, the Aedes species, and have overlapping geographical distributions.[9] They share remarkably similar clinical presentations, with common symptoms reported as fever, headache, vomiting, rash, myalgia, and severe arthralgia. Strategic public health programs advocate for combined surveillance and control efforts.[9]
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