Title : Rapid, isothermal detection of Zika virus: a potential alternative to RT-PCR
Abstract:
Zika virus (ZIKV) is a re-emerging flavivirus that poses a significant public health threat. ZIKV exhibits a wide array of non-vector borne human transmission routes, such as sexual transmission, transplacental transmission and blood transfusion. Detection and surveillance of ZIKV is considered paramount in prevention of major outbreaks. With the majority of cases reported in low-resource locations, simple, low-cost detection methods are considered highly desirable. Here we have developed a sensitive and specific ZIKV diagnostic using reverse transcription recombinase-aided amplification (RT-RAA) coupled with lateral flow strip detection (LFD) targeting the ZIKV NS1 gene. This diagnostic provides high throughput performance and speed without compromising sensitivity and specificity, compared to the gold-standard RT-PCR.
We show our ZIKV RT-RAA-LFD can detect 500 copies of ZIKV RNA/µL in under 30 minutes. Specificity testing confirmed that our assay does not detect any co-circulating flaviviruses (Dengue, West Nile, Japanese encephalitis and Yellow Fever viruses) or Chikungunya virus. Furthermore, the sample processing employed here results in complete inactivation of ZIKV (MR766 strain) in 5 minutes at RT. In comparison to conventional RT-PCR our ZIKV RT-RAA-LFD does not require expensive machinery, specialised laboratory settings or extensively trained personnel. Current research endeavours are focused on implementation of a one-step sample processing method coupled with our ZIKV RT-RAA-LFD of clinical samples (serum and urine) to assess the rapid, POC usability of our diagnostic in resource limited settings.
Collectively, our data suggests that our ZIKV RT-RAA-LFD diagnostic has the potential to become a clinically sensitive and specific alternative to RT-PCR, particularly within resource-limited settings, where ZIKV and several other arboviruses are endemic.
Keywords:
Recombinase Aided Amplification (RAA), Zika Virus (ZIKV), Lateral Flow Strip Detection (LFD), Point-of-care (POC).
Audience Take Away:
- Viral infection diagnostic technique that is different to conventional methods.
- Point-of-Care detection, diagnostics and surveillance of viruses and viral infections.
- Emerging arboviruses and viruses of concern.
