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Multiplex single-cell droplet PCR with machine learning for detection of high-risk human papillomaviruses.

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Title: Multiplex single-cell droplet PCR with machine learning for detection of high-risk human papillomaviruses.
Authors: Huang, Yizheng1,2 (AUTHOR), Sun, Linjun1,2,3 (AUTHOR), Liu, Wenwen1 (AUTHOR), Yang, Ling1 (AUTHOR), Song, Zhigang2,4 (AUTHOR), Ning, Xin2,3 (AUTHOR), Li, Weijun2,3 (AUTHOR), Tan, Manqing1,2 (AUTHOR), Yu, Yude1,2,5 (AUTHOR), Li, Zhao1,2,5 (AUTHOR) zhaoli@semi.ac.cn
Source: Analytica Chimica Acta. Apr2023, Vol. 1252, pN.PAG-N.PAG. 1p.
Subjects: Microfluidics, Human papillomavirus, Machine learning, Polymerase chain reaction, HeLa cells
Abstract: High-risk human papillomavirus (HPV) testing can significantly decline the incidence and mortality of cervical cancer. Microfluidic technology provides an effective method for accurate detection of high-risk HPV by utilizing multiplex single-cell droplet polymerase chain reaction (PCR). However, current strategies are limited by low-integration microfluidic chip, complex reagent system, expensive detection equipment and time-consuming droplet identification. Here, we developed a novel multiplex droplet PCR method that directly detected high-risk HPV sequences in single cells. A multiplex microfluidic chip integrating four flow-focusing structures was designed for one-step and parallel droplet preparation. Using single-cell droplet PCR, multi-target sequences were detected simultaneously based on a monochromatic fluorescence signal. We applied machine learning to automatically identify the large populations of single-cell droplets with 97% accuracy. HPV16, 18 and 45 sequences were sensitively detected without cross-contamination in mixed CaSki and Hela cells. The approach enables rapid and reliable detection of multi-target sequences in single cells, making it powerful for investigating cellular heterogeneity related to cancer diagnosis and treatment. [Display omitted] • A novel multiplex microfluidic chip developed for one-step droplet preparation. • High-risk HPV detection using multiplex single-cell droplet PCR. • Machine learning applied for automatic droplet classification and quantification. [ABSTRACT FROM AUTHOR]
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Database: Engineering Source
Description
Abstract:High-risk human papillomavirus (HPV) testing can significantly decline the incidence and mortality of cervical cancer. Microfluidic technology provides an effective method for accurate detection of high-risk HPV by utilizing multiplex single-cell droplet polymerase chain reaction (PCR). However, current strategies are limited by low-integration microfluidic chip, complex reagent system, expensive detection equipment and time-consuming droplet identification. Here, we developed a novel multiplex droplet PCR method that directly detected high-risk HPV sequences in single cells. A multiplex microfluidic chip integrating four flow-focusing structures was designed for one-step and parallel droplet preparation. Using single-cell droplet PCR, multi-target sequences were detected simultaneously based on a monochromatic fluorescence signal. We applied machine learning to automatically identify the large populations of single-cell droplets with 97% accuracy. HPV16, 18 and 45 sequences were sensitively detected without cross-contamination in mixed CaSki and Hela cells. The approach enables rapid and reliable detection of multi-target sequences in single cells, making it powerful for investigating cellular heterogeneity related to cancer diagnosis and treatment. [Display omitted] • A novel multiplex microfluidic chip developed for one-step droplet preparation. • High-risk HPV detection using multiplex single-cell droplet PCR. • Machine learning applied for automatic droplet classification and quantification. [ABSTRACT FROM AUTHOR]
ISSN:00032670
DOI:10.1016/j.aca.2023.341050