Rapid, point-of-care antigen and molecular tests base on CRISPR for diagnosis of HIV-1 infection

Background: Human immunodeficiency virus type one (HIV-1) is the leading cause of acquired immunodeficiency syndrome (AIDS). AIDS remains a global public health concern but can be effectively suppressed by life-long administration of combination antiretroviral therapy. Early detection and diagnosis are two key strategies for the prevention and control of HIV/AIDS. Rapid and accurate point-of-care testing (POCT) provides critical tools for managing HIV-1 epidemic in high-risk areas and populations. Methods: In this study, a POCT for HIV-1 RNA was developed by CRISPR/Cas13a lateral flow strip combined with reverse transcriptase recombinase-aided amplification (RT-RAA) technology, the results can be directly observed by naked eyes. Results: Moreover, with the degenerate base-binding CRISPR/Cas13a system was introduced into the RT-RAA primer designing, the technology developed in this study can be used to test majority of HIV-1 circulating strains with sensitivity of 1 copy/μL, while no obvious cross-reaction with other pathogens. We evaluated the sensitivity of this method for detecting HIV -1 RNA of clinical samples, the results showed that the sensitivity was 91.81% (101/110) and the specificity was 100% (48/48), the agreement rate between groups was 94.3%, the limit of detection (LOD) was 112 copies/mL. Conclusion: Above all, this method provides a point-of-care detection of HIV-1 RNA, which is stable, simple and with good sensitivity and specificity. This method has potential to be developed for promoting early diagnosis and treatment effect monitoring of HIV patients in clinical.