A Cas12a-based fluorescent microfluidic system for rapid on-site human papillomavirus diagnostics

文献类型： 外文期刊

第一作者： Huang, Baicheng

作者： Huang, Baicheng;Zeng, Zihan;Kan, Xingchi;Wu, Yue;Guo, Ling;Huang, Xingxu;Lou, Yufeng;Lou, Yufeng;Lou, Yufeng;Shi, Xinping;Wang, Muzhen;Tian, Xuemei;Wang, Xinjie

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关键词： Human papillomavirus; Diagnostic; Recombinase polymerase amplification; Cas12a; Microfluidics

期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：5.0； 五年影响因子：5.2 ）

ISSN： 0175-7598

年卷期： 2023 年 107 卷 20 期

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收录情况： SCI

摘要： Persistent infection with human papillomavirus (HPV) is the leading cause of cervical cancer, and early diagnosis is crucial for clinical management. However, the easy and rapid on-site diagnostic for HPV genotyping remains challenging. Here, we develop a Cas12a-based fluorescent microfluidic detection system for diagnosing six HPV subtypes (HPV6, HPV11, HPV16, HPV18, HPV31, and HPV33). A panel of crRNAs and recombinase polymerase amplification ( RPA) primers targeting the HPV L1 gene was screened for sensitive and specific detection. Furthermore, a one-pot RPA reaction was developed to amplify the six HPV subtypes without cross-reactivity. For on-site detection, we integrated the RPA-Cas12a detection into a microfluidic device, enabling the detection of processed clinical samples within 35 minutes. The assay was validated using 112 clinical swab samples and obtained consistent results with the qPCR assay, with a concordance rate of 99.1%. Overall, our diagnostic method offers a rapid, sensitive, and easy-to-use on-site assay for detecting HPV genotypes and holds promise for improving cervical cancer screening and prevention.

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