Abstract: Objective Zika virus (ZIKV) is an emerging arthropod-borne flavivirus transmitted by Aedes mosquitoes. Recent commentaries regarding ZIKV routes of transmission describe a potential transmission by transfusion. The current approaches in use are low in diagnostic efficiency and coverage area of ZIKV infection. Hence, this study intended to drive the ZIKV infection detection to effectively adaptable for any small to medium-sized laboratory or field survey. Methods We established, optimized, and evaluated a colloidal gold test strip for detection of ZIKV RNA based on CRISPR/Cas13a combined with reverse-transcription recombinase-aided amplification assay (RT-RAA) technology. Results The strip detection of ZIKV RNA was established based on RT-RAA-CRISPR-Cas13a technology with a sensitivity of 4 copies/μL and a specificity of 100%. ZIKV RNA gradient concentration standards and clinical samples were effectively identified by this approach. The positive coincidence rate for ZIKV was 100%, while the negative coincidence rate was 100%. The sensitivity, specificity, positive predictive agreement (PPA) and negative predictive agreement (NPA) values of Zika virus liquid plasma detection were 100%, 100%, 100%, and 100%, respectively. Conclusion We have developed rapid and portable RT-RAA-CRISPR/Cas13a-based strip of ZIKV RNA detection for patients. This study provides a visual and faster alternative to current PCR-based diagnosis for ZIKA infection.

Key words: CRISPR/Cas13a, RT-RAA, ZIKV, point of care test