A rapid isothermal CRISPR-Cas13a diagnostic test for genital herpes simplex virus infection

Xiaona Yin; Hao Luo; Han Zhou; Ziyan Zhang; Yinyuan Lan; Zhanqin Feng; Wentao Chen; Heping Zheng

doi:10.1016/j.isci.2023.108581

A rapid isothermal CRISPR-Cas13a diagnostic test for genital herpes simplex virus infection

iScience. 2023 Nov 27;27(1):108581. doi: 10.1016/j.isci.2023.108581. eCollection 2024 Jan 19.

Authors

Xiaona Yin^{1

2}, Hao Luo^{1

2}, Han Zhou^{1

2}, Ziyan Zhang^{1

2}, Yinyuan Lan^{1

2}, Zhanqin Feng^{1

2}, Wentao Chen^{1

2

3}, Heping Zheng^{1

2}

Affiliations

¹ Dermatology Hospital, Southern Medical University, Guangzhou 510091, China.
² Guangzhou Key Laboratory for Sexually Transmitted Diseases Control, Guangzhou 510091, China.
³ Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou 510515, China.

Abstract

Prompt diagnosis is essential for managing herpes simplex virus types 1 and 2 (HSV-1/2). Existing diagnostic methods are not widely available that required expensive or additional equipment for conducting examinations and result readouts, which can limit their utility in resource-constrained settings. We successfully developed a CRISPR-Cas13a-based assay for the detection and genotyping of HSV. Our assay demonstrated a high sensitivity of 96.15% and 95.15% for HSV-1 and HSV-2, respectively, with a specificity of 100% compared to a commercial qPCR assay when tested on 194 clinical samples. Remarkably, the assay enables a limit of detection of 1 copy/μL of viral DNA, facilitated by an enhanced input of RPA product and is designed for both mobile app integration and colorimetric interpretation, allowing for semiquantitative readings. These findings highlight the excellent performance of our CRISPR-based diagnostic in detecting HSV and its potential for point-of-care testing in resource-constrained settings.

Keywords: Diagnostics; Genetic engineering; Public health.