Rapid and multi-target genotyping of Helicobacter pylori with digital microfluidics.

Autor: Liu J; Department of Laboratory Medicine, Affiliated Hospital of Zunyi Medical University, Zunyi, 563006, China; College of Laboratory Medicine, Zunyi Medical University, Zunyi, 563000, China., Fu R; School of Medical Technology, Beijing Institute of Technology, Beijing, 100081, China; Engineering Research Center of Integrated Acousto-opto-electronic Microsystems, Ministry of Education of China, Beijing, 100081, China., Zhang S; School of Integrated Circuits and Electronic, Beijing Institute of Technology, Beijing, 100081, China; Engineering Research Center of Integrated Acousto-opto-electronic Microsystems, Ministry of Education of China, Beijing, 100081, China. Electronic address: shuailong.zhang@bit.edu.cn., Hou J; School of Medical Technology, Beijing Institute of Technology, Beijing, 100081, China., Ma H; CAS Key Laboratory of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, 215163, China., Hu S; CAS Key Laboratory of Bio-Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, 215163, China., Li H; School of Medical Technology, Beijing Institute of Technology, Beijing, 100081, China; Engineering Research Center of Integrated Acousto-opto-electronic Microsystems, Ministry of Education of China, Beijing, 100081, China., Zhang Y; Department of Gastroenterology, China-Japan Friendship Hospital, Beijing, 100029, China., Wang W; Department of Gastroenterology, The Third Medical Center of People's Liberation Army (PLA) General Hospital, Beijing, 100039, China., Qiao B; Beijing Anzhen Hospital, Capital Medical University, Beijing, 100029, China; Beijing Institute of Heart, Lung and Vascular Diseases, Key Laboratory of Remodeling-Related Cardiovascular Diseases, Beijing, 100029, China., Zang B; Zhejiang Anji GeneDetective Medical Technology Co. Ltd., Anji, 313300, China., Min X; Department of Laboratory Medicine, Affiliated Hospital of Zunyi Medical University, Zunyi, 563006, China; College of Laboratory Medicine, Zunyi Medical University, Zunyi, 563000, China., Zhang F; Department of Laboratory Medicine, Affiliated Hospital of Zunyi Medical University, Zunyi, 563006, China; College of Laboratory Medicine, Zunyi Medical University, Zunyi, 563000, China., Du J; Beijing Anzhen Hospital, Capital Medical University, Beijing, 100029, China; Beijing Institute of Heart, Lung and Vascular Diseases, Key Laboratory of Remodeling-Related Cardiovascular Diseases, Beijing, 100029, China. Electronic address: jiedu@ccmu.edu.cn., Yan S; Department of Laboratory Medicine, Affiliated Hospital of Zunyi Medical University, Zunyi, 563006, China; College of Laboratory Medicine, Zunyi Medical University, Zunyi, 563000, China. Electronic address: yanshengkai286@zmu.edu.cn.
Jazyk: angličtina
Zdroj: Biosensors & bioelectronics [Biosens Bioelectron] 2024 Jul 15; Vol. 256, pp. 116282. Date of Electronic Publication: 2024 Apr 11.
DOI: 10.1016/j.bios.2024.116282
Abstrakt: Helicobacter pylori (H. pylori) infection correlates closely with gastric diseases such as gastritis, ulcers, and cancer, influencing more than half of the world's population. Establishing a rapid, precise, and automated platform for H. pylori diagnosis is an urgent clinical need and would significantly benefit therapeutic intervention. Recombinase polymerase amplification (RPA)-CRISPR recently emerged as a promising molecular diagnostic assay due to its rapid detection capability, high specificity, and mild reaction conditions. In this work, we adapted the RPA-CRISPR assay on a digital microfluidics (DMF) system for automated H. pylori detection and genotyping. The system can achieve multi-target parallel detection of H. pylori nucleotide conservative genes (ureB) and virulence genes (cagA and vacA) across different samples within 30 min, exhibiting a detection limit of 10 copies/rxn and no false positives. We further conducted tests on 80 clinical saliva samples and compared the results with those derived from real-time quantitative polymerase chain reaction, demonstrating 100% diagnostic sensitivity and specificity for the RPA-CRISPR/DMF method. By automating the assay process on a single chip, the DMF system can significantly reduce the usage of reagents and samples, minimize the cross-contamination effect, and shorten the reaction time, with the additional benefit of losing the chance of experiment failure/inconsistency due to manual operations. The DMF system together with the RPA-CRISPR assay can be used for early detection and genotyping of H. pylori with high sensitivity and specificity, and has the potential to become a universal molecular diagnostic platform.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024. Published by Elsevier B.V.)
Databáze: MEDLINE
Externí odkaz: