A rapid isothermal CRISPR‒Cas13a diagnostic test for genital herpes simplex virus infection: associated data

Early diagnosis plays a pivotal role in the effective management of herpes simplex virus type 1 and 2 (HSV-1/2) infection. However, 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. Here, we successfully developed a CRISPR-based dual-target diagnostic system for the detection and genotyping of HSV-1 and HSV-2. Our assay demonstrated a high sensitivity of 96.15% and 94.85% for HSV-1 and HSV-2, respectively, with a specificity of 100% compared to a commercial qPCR assay when tested on 188 clinical samples. Furthermore, we have adapted our assay to be used with a mobile app or naked eyes-based colorimetric readout, which showed a limit of detection of 1 copy/μL for both HSV-1 and HSV-2 DNA when increased input of the RPA product was used. 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.

早期诊断在1型和2型单纯疱疹病毒（Herpes Simplex Virus Type 1 and 2, HSV-1/2）感染的有效管理中发挥关键作用。然而，现有诊断方法需配备昂贵或额外设备以开展检测与结果判读，尚未得到广泛应用，这限制了其在资源匮乏地区的实用性。本研究成功构建了一套基于CRISPR（Clustered Regularly Interspaced Short Palindromic Repeats）的双靶点诊断系统，用于HSV-1与HSV-2的检测及基因分型。在188份临床样本中开展验证时，相较于商用qPCR（定量聚合酶链式反应，quantitative Polymerase Chain Reaction）检测体系，本检测方法对HSV-1和HSV-2的检测灵敏度分别达96.15%与94.85%，特异性则为100%。此外，本研究将该检测体系适配为可通过移动应用或肉眼比色法进行结果判读；在提升RPA（重组酶聚合酶扩增，Recombinase Polymerase Amplification）产物投入量的条件下，该体系对HSV-1和HSV-2 DNA的检测限均达到1拷贝/微升。上述研究结果凸显了本CRISPR基诊断系统在HSV检测中的优异性能，以及其在资源匮乏地区开展即时检测（Point-of-Care Testing, POCT）的应用潜力。