Rapid and simultaneous visual typing of high-risk HPV-16/18 with use of integrated lateral flow strip platform,Microchimica Acta

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Rapid and simultaneous visual typing of high-risk HPV-16/18 with use of integrated lateral flow strip platform
Microchimica Acta ( IF 5.3 ) Pub Date : 2022-08-25 , DOI: 10.1007/s00604-022-05449-5
Qi Chen ₁ , Li Yao ₁ , Qian Wu ₁ , Jianguo Xu ₁ , Chao Yan ₁ , Chuanxiang Guo ₂ , Chao Zhang ₂ , Tao Xu ₃ , Panzhu Qin _{1,

3} , Wei Chen _{1,

4}

Affiliation

A biosensor for rapid and simultaneous visual identification of high-risk human papillomavirus (HPV) genotypes 16 and 18 in clinical samples based on polymerase chain reaction (PCR) integrated lateral flow strip platform was developed. Using an one-step protocol to extract nucleic acid rapidly and the functionalized primer sets specific to HPV-16 and 18 were designed for the simultaneous amplification. In the presence of target HPV genotypes, the corresponding functionalized primer sets will participate in the PCR process and produce numerous duplex functionalized dsDNA amplicons. With the bridge effect of duplex functionalized dsDNA amplicons between gold nanoparticles-fluorescein isothiocyanate antibody conjugates (AuNP-FITC antibody conjugates) and other two antibodies on corresponding test line (T₁ or T₂), visualized color signals on test lines could be obtained directly visible with a naked eye. Combining the high amplification efficiency of PCR and the visualized sensing of LFS, as low as 700 copies of HPV-16 and 18 DNA were detected simultaneously within 75 min, which can promote application in the resource limited settings.

Graphical abstract

High-risk genotypes of HPV-16 and HPV-18 were easily and simultaneously screened with the amplification-assisted molecular lateral flow strip by on-site observation in the resource-limited settings.

中文翻译：

使用集成侧向流动条平台对高危 HPV-16/18 进行快速、同步的视觉分型

开发了一种基于聚合酶链反应 (PCR) 集成侧向流动条带平台的生物传感器，用于快速同时视觉识别临床样本中高危人乳头瘤病毒 (HPV) 基因型 16 和 18。使用一步法快速提取核酸，并设计了对 HPV-16 和 18 特异的功能化引物组用于同时扩增。在存在目标 HPV 基因型的情况下，相应的功能化引物组将参与 PCR 过程并产生大量双链功能化 dsDNA 扩增子。在金纳米粒子-异硫氰酸荧光素抗体偶联物（AuNP-FITC抗体偶联物）和相应检测线（T ₁或T₂ )，可以用肉眼直接看到测试线上的可视化颜色信号。结合 PCR 的高扩增效率和 LFS 的可视化传感，在 75 分钟内同时检测到低至 700 拷贝的 HPV-16 和 18 DNA，可促进在资源有限的环境中的应用。