Abstract
Zika virus (ZIKV) has gained global attention as an etiologic agent of fetal microcephaly and Guillain-Barré syndrome. Existing immuno-based rapid tests often fail to distinguish between Zika and related flaviviruses that are common in affected regions of Central and South Americas and the Caribbean. The US CDC and qualified state health department laboratories can perform the reverse transcription polymerase chain reaction (RT-PCR) ZIKV test using highly sophisticated instruments with long turnaround times. The preliminary results of a portable and low-cost molecular diagnostics system for ZIKV infection are reported here. In less than 15 minutes, this low-cost platform can automatically perform high quality RNA extraction from up to 12 ZIKV-spiked urine samples simultaneously. It can also perform reverse transcription recombinase polymerase amplification reaction (RT-RPA) in ≤15 minutes. The fluorescent signal produced from probe-based RT-RPA or RT-PCR assays can be monitored using LEDs and a smartphone camera. In addition, the RT-RPA and RT-PCR assays do not cross-react with dengue and chikungunya viral RNA. This low-cost system lacks complicated, sensitive and high cost components, making it suitable for resource-limited settings. It has the potential to offer simple sample-to-answer molecular diagnostics and can inform healthcare workers of patients’ diagnosis promptly.
Highlights
IntroductionZika virus (ZIKV) is an emerging mosquito-borne pathogen (family Flaviviridae, genus Flavivirus) that was first isolated in 1947 from a rhesus monkey in Uganda’s Zika forest[1]
Zika virus (ZIKV) is an emerging mosquito-borne pathogen that was first isolated in 1947 from a rhesus monkey in Uganda’s Zika forest[1]
The molecular detection of viral RNA in serum using a reverse transcription polymerase chain reaction (RT-PCR) protocol described by Lanciotti et al during investigations of the ZIKV outbreak on Yap Island has been deemed suitable for acute phase diagnosis of ZIKV10,14–17
Summary
Zika virus (ZIKV) is an emerging mosquito-borne pathogen (family Flaviviridae, genus Flavivirus) that was first isolated in 1947 from a rhesus monkey in Uganda’s Zika forest[1]. These recent studies are in agreement with previous reports that other flavivirus genomes, such as those of dengue[21], West Nile[22] and most recently ZIKV RNA23 can be detected in urine longer than in serum These reports suggest that urine samples are a superior choice for diagnosis of ZIKV infection due to the higher RNA load and longer duration compared to serum. For these reasons, urine was chosen in this study as the sample specimen to demonstrate the diagnostic utility of the novel platform for detection of ZIKV RNA using isothermal amplification techniques such as real-time reverse transcription recombinase polymerase amplification (RT-RPA) and the more traditional real-time RT-PCR. For the detection of RNA, reverse-transcriptase is added so “one-step” RT-RPA can be used[30,31,32,33,34,35]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
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 call
