CRISPR-Cas12a based HSV DNA detection method using quantum dot-labeled immunochromatographic strips

Abstract

Herpes simplex virus (HSV) infection poses a significant public health challenge, particularly affecting vulnerable populations such as pregnant women, newborns, and immunocompromised individuals. Traditional diagnostic methods for HSV are often time-consuming, requiring specialized expertise and infrastructure, resulting in delays and limitations in resource-limited regions. In this research, we established a novel detection method that integrates CRISPR-Cas12a with Recombinase Assisted Amplification (RAA), enabling the sensitive and specific detection of HSV DNA. Compared to TaqMan PCR, our CRISPR-Cas12a-based lateral flow assay (CRI-LFA) using universal primers demonstrated a sensitivity of 90.38 %, specificity of 98.08 %, positive predictive value (PPV) of 100 %, negative predictive value (NPV) of 91.22 %, kappa value of 0.904, and an area under the receiver operating characteristic curve (AUC) of 0.973. The subsequent typing tests on our platform for HSV-1 and HSV-2 showed a sensitivity of 100 % (95 % CI: 74.12 %–100 %) and 95.83 % (95 % CI: 86.02 %–99.26 %), respectively. Specificity was 97.85 % (95 % CI: 92.49 %–99.62 %) for HSV-1 and 96.64 % (95 % CI: 85.39 %–98.54 %) for HSV-2. The kappa values were 0.906 for HSV-1 and 0.883 for HSV-2, with PPV of 84.6 % and 97.7 %, and NPV of 100 % and 91.7 %, respectively. Our portable paper strip assay provides a convenient and user-friendly testing option suitable for decentralized settings. This approach has the potential to facilitate early detection and enhance public health efforts in controlling HSV transmission.