Xi Xuxiang, Cao Wen-Ling, Yao Xuebing, Chen Jie, Huang Defa, Yang Tong, Liu Zhaoxia, Xie Wei, Xia Yu, Zhong Tianyu
Department of Laboratory Medicine, First Affiliated Hospital of Gannan Medical University, Ganzhou, Jiangxi Province, China.
Department of Clinical Laboratory, Guangzhou Institute of Dermatology, Guangzhou, Guangdong Province, China.
Mol Cell Probes. 2022 Feb;61:101787. doi: 10.1016/j.mcp.2021.101787. Epub 2021 Dec 4.
Current human papillomavirus (HPV) detection methods require complex instruments, skilled staff and have a high cost. Therefore, novel testing approaches are needed which are easy to implement, highly sensitive, and low cost. Loop-mediated isothermal amplification (LAMP) is an isothermal amplification technique. In this study, according to the conditions in China, a novel LAMP method for detecting seven high-risk HPV subtypes (16, 18, 33, 39, 45, 52, and 58) was designed and evaluated. The DNA from plasmid and cervical specimens was extracted using Chelex 100 and measured by qPCR and LAMP assay. LAMP products were observed under ultraviolet light. HPV sequences were successfully amplified and a plateau time of 19-75 min was maintained. The concentration of positive reactions ranged between 20 copies/μL and 200000 copies/μL. Additionally, there was no cross-reactivity between HPV16, 18, 33, 39, 45, 52, 58, 31, 35, 45, 51, 56, 59, 66, or 68. For clinical samples, the LAMP assay had high sensitivity and specificity for HPV16, 18, 33, 39, 45, 52, and 58. However, 5% (72/1447) of the samples tested yielded false-positive results. In conclusion, the novel LAMP assay for HPV16, 18, 33, 39, 45, 52, and 58 has high sensitivity and specificity, a low cost, and is simple and rapid to perform. The LAMP assay can improve HPV detection in resource-limited settings, especially in primary care hospitals and rural areas.
目前的人乳头瘤病毒（HPV）检测方法需要复杂的仪器、技术熟练的工作人员，且成本高昂。因此，需要易于实施、高度灵敏且低成本的新型检测方法。环介导等温扩增技术（LAMP）是一种等温扩增技术。在本研究中，根据中国的实际情况，设计并评估了一种用于检测七种高危HPV亚型（16、18、33、39、45、52和58）的新型LAMP方法。使用Chelex 100从质粒和宫颈标本中提取DNA，并通过定量聚合酶链反应（qPCR）和LAMP检测进行测量。在紫外线下观察LAMP产物。HPV序列成功扩增，平台期维持在19至75分钟。阳性反应的浓度范围为20拷贝/微升至200000拷贝/微升。此外，HPV16、18、33、39、45、52、58与31、35、45、51、56、59、66或68之间无交叉反应。对于临床样本，LAMP检测对HPV16、18、33、39、45、52和58具有高灵敏度和特异性。然而，5%（72/1447）的检测样本产生了假阳性结果。总之，用于检测HPV16、18、33、39、45、52和58的新型LAMP检测具有高灵敏度和特异性、低成本，且操作简单快速。LAMP检测可改善资源有限环境中的HPV检测，特别是在基层医院和农村地区。
