Yin Xiaona, Luo Hao, Zhou Han, Zhang Ziyan, Lan Yinyuan, Feng Zhanqin, Chen Wentao, Zheng Heping
Dermatology Hospital, Southern Medical University, Guangzhou 510091, China.
Guangzhou Key Laboratory for Sexually Transmitted Diseases Control, Guangzhou 510091, China.
iScience. 2023 Nov 27;27(1):108581. doi: 10.1016/j.isci.2023.108581. eCollection 2024 Jan 19.
Prompt diagnosis is essential for managing 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.
及时诊断对于管理1型和2型单纯疱疹病毒（HSV - 1/2）至关重要。现有的诊断方法并不广泛适用，因为这些方法需要昂贵的设备或额外的设备来进行检测和结果读取，这可能会限制它们在资源有限环境中的实用性。我们成功开发了一种基于CRISPR - Cas13a的检测方法，用于检测HSV并进行基因分型。在对194份临床样本进行检测时，与商业定量聚合酶链反应（qPCR）检测方法相比，我们的检测方法对HSV - 1和HSV - 2的敏感性分别高达96.15%和95.15%，特异性为100%。值得注意的是，该检测方法通过增加重组酶聚合酶扩增（RPA）产物的输入量，实现了1拷贝/微升病毒DNA的检测限，并且设计用于移动应用集成和比色解读，从而实现半定量读数。这些发现突出了我们基于CRISPR的诊断方法在检测HSV方面的卓越性能及其在资源有限环境中进行即时检测的潜力。
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