A microchip electrophoresis method for rapid nucleic acid extraction and bacteria detection

Yi Qiao; Lu Ding; Chao Huang; Yin Chen; Tian Wen; Jing Tu

doi:10.1016/j.talanta.2025.128493

A microchip electrophoresis method for rapid nucleic acid extraction and bacteria detection

Talanta. 2025 Jun 20:296:128493. doi: 10.1016/j.talanta.2025.128493. Online ahead of print.

Authors

Yi Qiao¹, Lu Ding¹, Chao Huang², Yin Chen², Tian Wen³, Jing Tu⁴

Affiliations

¹ State Key Laboratory of Digital Medical Engineering, School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096, China.
² NHC Key Laboratory of Enteric Pathogenic Microbiology, Jiangsu Provincial Medical Key Laboratory of Pathogenic Microbiology in Emerging Major Infectious Diseases, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, 210009, China.
³ NHC Key Laboratory of Enteric Pathogenic Microbiology, Jiangsu Provincial Medical Key Laboratory of Pathogenic Microbiology in Emerging Major Infectious Diseases, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, 210009, China. Electronic address: wentian@jscdc.cn.
⁴ State Key Laboratory of Digital Medical Engineering, School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096, China. Electronic address: jtu@seu.edu.cn.

PMID: 40570436
DOI: 10.1016/j.talanta.2025.128493

Abstract

The human respiratory system is highly susceptible to bacterial infections. Nucleic acid testing (NAT) strategies are of vital importance in identifying the presence of specific bacteria in samples. The process of nucleic acid hybridization ensures a high degree of specificity. The release and purification of DNA form the basis for NAT. Currently, magnetic beads and other absorption-based methods are the prevalent options for DNA extraction. Nevertheless, their low integration and high equipment demands restrict their application. In this study, we present a microchip electrophoresis detection (MED) platform for integrated bacterial DNA extraction based on gradient-gel electrophoresis. Bacteria are lysed directly on the chip and the released DNA is subsequently purified through electrophoresis. Each step of the extraction process was systematically evaluated and the entire system was put into practical use. When combined with downstream quantitative polymerase chain reaction (qPCR), this system exhibits high sensitivity in detection. The results are concordant with those of the conventional method at high target concentrations and the detection limit of the entire system prevails. Additionally, the chip is also compatible with on-chip loop-mediated isothermal amplification (LAMP), highlighting its substantial potential for future applications in rapid and efficient bacterial detection.

Keywords: Bacteria detection; DNA extraction; Microchip electrophoresis; Point-of-care testing.