Abstract
BackgroundThe recent emergence of Zika virus (ZIKV) in Brazil and its precipitous expansion throughout the Americas has highlighted the urgent need for a rapid and reliable on-site diagnostic assay suitable for viral detection. Such point-of-need (PON), low-cost diagnostics are essential for ZIKV control in vulnerable areas with limited resources.MethodsWe developed and evaluated a ZIKV-specific field-deployable RT-iiPCR reagent set targeting the E gene for rapid detection of ZIKV in ZIKV-spiked human and mosquito specimens, and compared its performance to the Center for Disease Control and Prevention (CDC) and Pan American Health Organization (PAHO) RT-qPCR assays targeting the E and NS2B genes, respectively.ResultsThese assays demonstrated exclusive specificity for ZIKV (African and Asian lineages), had limits of detection ranging from 10 to 100 in vitro transcribed RNA copies/μl and detection endpoints at 10 plaque forming units/ml of infectious tissue culture fluid. Analysis of human whole blood, plasma, serum, semen, urine, and mosquito pool samples spiked with ZIKV showed an agreement of 90% (k = 0.80), 92% (k = 0.82), 95% (k = 0.86), 92% (k = 0.81), 90% (k = 0.79), and 100% (k = 1), respectively, between the RT-iiPCR assay and composite results from the reference RT-qPCR assays. Overall, the concurrence between the ZIKV RT-iiPCR and the reference RT-qPCR assays was 92% (k = 0.83).ConclusionsThe ZIKV RT-iiPCR has a performance comparable to the reference CDC and PAHO RT-qPCR assays but provides much faster results (~1.5 h) with a field-deployable system that can be utilized as a PON diagnostic with the potential to significantly improve the quality of the health care system in vulnerable areas.
Highlights
The recent emergence of Zika virus (ZIKV) in Brazil and its precipitous expansion throughout the Americas has highlighted the urgent need for a rapid and reliable on-site diagnostic assay suitable for viral detection
ZIKV belongs to the family Flaviviridae, genus Flavivirus, and it is closely related to other mosquito-borne flaviviruses such as dengue (DENV), West Nile (WNV), and Japanese encephalitis viruses (JEV) [14, 15]
The analytical sensitivity of the PON ZIKV RT-insulated isothermal PCR (iiPCR) was determined using a (a) ten-fold dilution series of ZIKV in vitro transcribed (IVT) RNA (107 to 0.1 IVT RNA copies/μl) containing the target sequence, and (b) ten-fold serial dilutions (100–10−13) of nucleic acid extracted from Tissue culture fluid (TCF) derived from ZIKV PRVABC59infected Vero cells containing a viral titer of 107 PFU/ml
Summary
The recent emergence of Zika virus (ZIKV) in Brazil and its precipitous expansion throughout the Americas has highlighted the urgent need for a rapid and reliable on-site diagnostic assay suitable for viral detection. Such point-ofneed (PON), low-cost diagnostics are essential for ZIKV control in vulnerable areas with limited resources. ZIKV is primarily transmitted by Aedes species of mosquitoes, including the urban and suburban mosquito species A. aegypti and A. albopictus, implicated in the transmission of DENV and alphaviruses such as Chikungunya virus (CHIKV) [18, 21,22,23,24,25]. ZIKV can be potentially transmitted by blood transfusions [32,33,34,35,36], and transmission through transfusion of a platelet concentrate has been recently reported in Brazil [36]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
