Bio-manufacturing Engineering Laboratory, Tsinghua Shenzhen International Graduate School, Tsinghua University, Shenzhen, China.
Analyst. 2021 Aug 21;146(16):5102-5114. doi: 10.1039/d1an00623a. Epub 2021 Jul 15.
Human papillomavirus (HPV) causes the prevalent sexually transmitted infection that accounts for the majority of cervical cancer incidences. Therefore, the development of a rapid, accurate, automatic and affordable nucleic acid detection strategy is urgently required for HPV tests, among which microfluidic chip is a promising diagnostic method. In this work, we developed a microfluidic detection system consisting of a microfluidic chip and the corresponding detection equipment to diagnose high-risk HPV. The proposed method integrates nucleic acid purification, isothermal amplification and real-time fluorescence detection into one device. Moreover, it demonstrates good detection performance such as high specificity of primer sets (100%) and exceptional stability (coefficient of variation <6%) among five HPV genotypes. Besides, the microfluidic loop-mediated isothermal amplification (LAMP) assay is accurate (specificity of 91.7% and sensitivity of 100%) and fast (average time threshold = 10.56 minutes) when considering the conventional qPCR assay as the gold standard. The integrated microfluidic detection system offers automated and rapid diagnosis within 40 minutes and shows broad potential to deliver point-of-care detection in resource-limited circumstances owing to its simplicity and affordability.
人乳头瘤病毒（HPV）导致了常见的性传播感染，占大多数宫颈癌发病率的原因。因此，迫切需要开发一种快速、准确、自动和负担得起的核酸检测策略，用于 HPV 测试，其中微流控芯片是一种很有前途的诊断方法。在这项工作中，我们开发了一种由微流控芯片和相应的检测设备组成的微流控检测系统，用于诊断高危型 HPV。该方法将核酸纯化、等温扩增和实时荧光检测集成到一个装置中。此外，它表现出了良好的检测性能，如引物组的高特异性（100%）和五个 HPV 基因型之间的出色稳定性（变异系数<6%）。此外，微流控环介导等温扩增（LAMP）检测具有较高的准确性（特异性为 91.7%，灵敏度为 100%），与传统 qPCR 检测相比，速度较快（平均时间阈值=10.56 分钟）。作为金标准。集成的微流控检测系统可在 40 分钟内实现自动化和快速诊断，由于其简单性和可负担性，有望在资源有限的情况下实现即时检测。
