Chemical Engineering Research Center, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300350, P. R. China.
Tianjin Key Laboratory of Membrane Science and Desalination Technology, Tianjin University, Tianjin 300072, P. R. China.
Anal Methods. 2024 Nov 14;16(44):7557-7566. doi: 10.1039/d4ay01625d.
Human papillomavirus 16 (HPV16) infection is the leading cause of cervical cancer. The current mainstream method for detecting HPV16 is quantitative real-time PCR (qPCR). However, due to its time-consuming nature and reliance on expensive qPCR instruments, there is growing interest in more convenient, rapid and private approaches such as recombinase polymerase amplification (RPA). In this study, we innovated an effective method for HPV16 detection by integrating a RPA system with lateral flow strip (LFS) detection. Primers with optimal efficiency and specificity were designed, and false positive signals caused by dimers were eliminated by reducing the probe concentration. The RPA-LFS method demonstrates high sensitivity and specificity, capable of detecting 230 copies per reaction of HPV16 within 25 min without cross-reactivity to other subtypes. It exhibits good tolerance, remaining unaffected by 1.0% miconazole, 0.5% tioconazole and 1.0% hemoglobin. The results of clinical samples detected by this method were consistent with those of qPCR. The method provides a practical reference for HPV16 diagnosis and can be valuable in home and resource-limited settings, contributing to the reduction of cervical cancer incidence.
人乳头瘤病毒 16（HPV16）感染是宫颈癌的主要病因。目前检测 HPV16 的主流方法是定量实时 PCR（qPCR）。然而，由于其耗时且依赖昂贵的 qPCR 仪器，因此人们越来越感兴趣于更方便、快速和私密的方法，如重组酶聚合酶扩增（RPA）。在这项研究中，我们通过将 RPA 系统与横向流动条（LFS）检测相结合，创新了一种有效的 HPV16 检测方法。设计了具有最佳效率和特异性的引物，并通过降低探针浓度消除了由二聚体引起的假阳性信号。RPA-LFS 方法具有高灵敏度和特异性，能够在 25 分钟内检测到每个反应 230 个拷贝的 HPV16，并且与其他亚型无交叉反应。它具有良好的耐受性，不受 1.0%咪康唑、0.5%噻康唑和 1.0%血红蛋白的影响。该方法检测临床样本的结果与 qPCR 一致。该方法为 HPV16 诊断提供了实用参考，可在家庭和资源有限的环境中具有价值，有助于降低宫颈癌的发病率。
