Highly sensitive and rapid point-of-care testing for HIV-1 infection based on CRISPR-Cas13a system

Abstract

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 RNA with limit of detection (LOD) 1 copy/μL, while no obvious cross-reaction with other pathogens. We evaluated this method for detecting HIV-1 RNA of clinical samples, the results showed that the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy were 91.81% (85.03- 96.19%), 100% (92.60–100%), 100% (96.41–100%), 39.14% (25.59–54.60%) and 92.22% (86.89–95.88%), respectively. The lowest viral load detectable by this method was 112copies/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.

Electro‐Optical Multiclassification Platform for Minimizing Occasional Inaccuracy in Point‐of‐Care Biomarker Detection

“…Thus, our technology exhibited remarkable diagnostic accuracy and accessibility in point-of-care settings, which are greater than those of existing laboratory tests and commercial kits (Figure 4d). [6,13,15,20,36,39]…”

“…[7,9,[36][37][38] At 100% statistical power (>150 clinical samples), the accuracy will be traded-off with a value below 93% for minutelevel POCT diagnosis, especially in on-site scenes with multivariable interference. [5,10,28,31,[39][40][41] Therefore, it is highly desirable to develop a comprehensive platform that minimizes occasional inaccuracies in complex on-site settings within a few minutes. [17,[41][42][43] In this context, we describe a dual-mode multiclassification diagnostic platform (termed DMDP, Figure 1) based on a microfluidic chip that combines electrochemiluminescence (ECL) and field-effect transistor (FET) sensing units.…”