Reproductive Medical Center, Department of Obstetrics and Gynecology, Seventh Medical Center of PLA General Hospital, Beijing 100700, China.
State Key Laboratory of Space Medicine, China Astronaut Research and Training Center, Beijing 100094, China.
Langmuir. 2024 Aug 13;40(32):16722-16730. doi: 10.1021/acs.langmuir.4c00793. Epub 2024 Aug 2.
Cervical cancer is among the most common malignant tumors in women. The development of rapid screening techniques plays an important role in early screening for cancer treatment. We have developed an HPV screening method, which effectively combines the high-efficiency nucleic acid enrichment of chitosan-modified filter paper and the rapid visual detectability of colorimetric LAMP, along with the enhancement of the tolerance ability of the pH-sensitive LAMP reagent to acidic original samples, making the detection of HPV 16/18 easy to carry out and reliable, which is helpful for the epidemiological prevention and control strategies of HPV-induced cancer. This technique can simultaneously exhibit the " amplification" capability of chitosan-modified filter paper and the nontemperature cycle dependence of visual LAMP detection. Therefore, DNA extraction and amplification can be performed efficiently and quickly within a single reaction where all DNA is concentrated in the QF paper disc. By embedding amino-modified filter paper into the plastic chip, a simple and reliable disposable chip was prepared for rapid HPV16 and HPV18 detection from clinical endometrial samples, and the results were 100% consistent with clinical diagnosis. More importantly, even after the sample was diluted 100-fold, HPV16/18-infected cells could be accurately identified, showing the advantages of the system in early cancer screening. Moreover, for endometrial samples containing plenty of cells, the filter paper could be used to enrich cells by filtration, preventing the acidic fluid from impacting pH-induced colorimetric LAMP detection and realizing direct amplification for HPV identification without nucleic acid extraction. This easy-to-operate system that can analyze a wide range of samples will be suitable for routine on-site HPV screening, dramatically extending the applications and utility for rapid, near-patient nucleic acid testing.
宫颈癌是女性最常见的恶性肿瘤之一。快速筛查技术的发展在癌症治疗的早期筛查中起着重要作用。我们开发了一种 HPV 筛查方法，该方法有效地将壳聚糖修饰滤纸的高效核酸富集与比色环介导等温扩增（LAMP）的快速可视化检测能力相结合，同时增强了 pH 敏感 LAMP 试剂对酸性原始样本的耐受能力，使 HPV 16/18 的检测变得容易且可靠，这有助于 HPV 诱导癌症的流行病学预防和控制策略。该技术可以同时展现壳聚糖修饰滤纸的“扩增”能力和可视化 LAMP 检测的非温度循环依赖性。因此，所有 DNA 都可以集中在 QF 滤纸盘上，在单个反应中可以高效快速地进行 DNA 提取和扩增。通过将氨基修饰的滤纸嵌入塑料芯片中，我们制备了一种简单可靠的一次性芯片，可用于从临床子宫内膜样本中快速检测 HPV16 和 HPV18，其结果与临床诊断完全一致。更重要的是，即使将样本稀释 100 倍，也可以准确识别 HPV16/18 感染的细胞，显示出该系统在早期癌症筛查中的优势。此外，对于含有大量细胞的子宫内膜样本，滤纸可以用于过滤富集细胞，防止酸性液体影响 pH 诱导的比色 LAMP 检测，并实现 HPV 鉴定的直接扩增，而无需核酸提取。这种易于操作的系统可以分析广泛的样本，将非常适合常规现场 HPV 筛查，极大地扩展了快速、近患者核酸检测的应用和实用性。
