A portable all-in-one microfluidic device with real-time colorimetric LAMP for HPV16 and HPV18 DNA point-of-care testing

Hao Bai; Yuqing Liu; Linbo Gao; Tao Wang; Xiaoli Zhang; Jie Hu; Lisha Ding; Yueting Zhang; Qian Wang; Lei Wang; Jianlong Li; Zhifeng Zhang; Yang Wang; Chenlan Shen; Binwu Ying; Xiaoyu Niu; Wenchuang Hu

doi:10.1016/j.bios.2023.115968

A portable all-in-one microfluidic device with real-time colorimetric LAMP for HPV16 and HPV18 DNA point-of-care testing

Biosens Bioelectron. 2024 Mar 15:248:115968. doi: 10.1016/j.bios.2023.115968. Epub 2023 Dec 22.

Authors

Hao Bai¹, Yuqing Liu², Linbo Gao³, Tao Wang², Xiaoli Zhang², Jie Hu⁴, Lisha Ding², Yueting Zhang⁵, Qian Wang², Lei Wang⁶, Jianlong Li⁶, Zhifeng Zhang⁶, Yang Wang⁷, Chenlan Shen⁸, Binwu Ying⁹, Xiaoyu Niu¹⁰, Wenchuang Hu¹¹

Affiliations

¹ Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, 610041, China; Med+X Center for Manufacturing, West China Hospital, Sichuan University, Chengdu, 610041, China.
² Department of Obstetrics and Gynecology, West China Second University Hospital, Sichuan University, Chengdu, 610041, China; Center for Translational Medicine, Key Laboratory of Birth Defects and Related Diseases of Women and Children, West China Second University Hospital, Sichuan University, Chengdu, 610041, China; Laboratory of Metabolomics and Gynecological Disease Research, West China Second University Hospital, Sichuan University, Chengdu, 610041, China.
³ Center for Translational Medicine, Key Laboratory of Birth Defects and Related Diseases of Women and Children, West China Second University Hospital, Sichuan University, Chengdu, 610041, China.
⁴ Med+X Center for Manufacturing, West China Hospital, Sichuan University, Chengdu, 610041, China; Precision Medicine Center, West China Hospital, Sichuan University, Chengdu, 610041, China.
⁵ Department of Obstetrics and Gynecology, West China Second University Hospital, Sichuan University, Chengdu, 610041, China; Center for Translational Medicine, Key Laboratory of Birth Defects and Related Diseases of Women and Children, West China Second University Hospital, Sichuan University, Chengdu, 610041, China.
⁶ One-Chip Biotechnology Co. Ltd, Chengdu, 610041, China.
⁷ Beijing Advanced Innovation Center for Biomedical Engineering, Key Laboratory for Biomechanics and Mechanobiology, School of Engineering Medicine, Beihang University, Beijing, 100083, China.
⁸ Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, 610041, China.
⁹ Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, 610041, China; Med+X Center for Manufacturing, West China Hospital, Sichuan University, Chengdu, 610041, China. Electronic address: yingbinwu@scu.edu.cn.
¹⁰ Department of Obstetrics and Gynecology, West China Second University Hospital, Sichuan University, Chengdu, 610041, China; Center for Translational Medicine, Key Laboratory of Birth Defects and Related Diseases of Women and Children, West China Second University Hospital, Sichuan University, Chengdu, 610041, China; Laboratory of Metabolomics and Gynecological Disease Research, West China Second University Hospital, Sichuan University, Chengdu, 610041, China. Electronic address: niuxy@scu.edu.cn.
¹¹ Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, 610041, China; Med+X Center for Manufacturing, West China Hospital, Sichuan University, Chengdu, 610041, China; Precision Medicine Center, West China Hospital, Sichuan University, Chengdu, 610041, China. Electronic address: huwenchuang@wchscu.cn.

PMID: 38150799
DOI: 10.1016/j.bios.2023.115968

Abstract

Screening for high-risk human papillomavirus (HPV) infection is one of the most important preventative measures for cervical cancer. However, fast, convenient, and low-cost HPV detection remains challenging, especially in resource-limited settings. Here, we report a portable all-in-one device (PAD) for point-of-care testing (POCT) for HPV16 and HPV18 DNA in cervical swabs. The PAD was engineered to integrate modules for extraction-free sample lysis, loop-mediated isothermal amplification (LAMP) with lyophilized reagent beads, and real-time colorimetric signal sensing into a single miniaturized device, considerably shortening the sample-to-result time to 15 min. The precision liquid handling in the completely sealed microfluidic chip is achieved by a uniquely designed pressure-balanced automatic liquid flow mechanism, thereby eliminating the need for manual manipulation of liquids and thus the risk of biohazards. The PAD employs an improved real-time colorimetric LAMP (rcLAMP) assay with a limit of detection (LOD) of 1 copy/μL, enabled by enhanced assay chemistry to maximize the reaction kinetics. To validate this device for clinical application, we tested 206 clinical cervical swab samples and obtained a sensitivity of 92.1% and a specificity of 99.0%. This custom PAD enabled by microfluidic and electronic engineering techniques can be configured for the simultaneous detection of HPV16 and HPV18 or other pathogens in point-of-care applications.

Keywords: Diagnostic device; Human papillomavirus; Microfluidics; Point-of-care testing; Real-time colorimetric LAMP.

MeSH terms

Biosensing Techniques*
Colorimetry / methods
DNA, Viral / genetics
Female
Human papillomavirus 16 / genetics
Human papillomavirus 18 / genetics
Humans
Lab-On-A-Chip Devices
Microfluidics
Nucleic Acid Amplification Techniques / methods
Papillomavirus Infections* / diagnosis
Point-of-Care Testing
Sensitivity and Specificity

Substances

DNA, Viral