A novel loop-mediated isothermal amplification-lateral flow dipstick method for Helicobacter pylori detection

Wenwen Liu; Gang Lu; Yu Wang; Zhenghong Chen; Yunyun Gao; Zhipeng Yin; Yi Wu; Xiaoqian Lv; Pengbo Guo; Yinghui Zhao

doi:10.3389/fmicb.2023.1094600

A novel loop-mediated isothermal amplification-lateral flow dipstick method for Helicobacter pylori detection

Front Microbiol. 2023 Mar 23:14:1094600. doi: 10.3389/fmicb.2023.1094600. eCollection 2023.

Authors

Wenwen Liu¹, Gang Lu^{2

3}, Yu Wang^{2

3}, Zhenghong Chen⁴, Yunyun Gao², Zhipeng Yin², Yi Wu², Xiaoqian Lv², Pengbo Guo^{2

3}, Yinghui Zhao^{2

3}

Affiliations

¹ Department of Clinical Laboratory Medicine, The First Affiliated Hospital of Shandong First Medical University and Shandong Provincial Qianfoshan Hospital, Shandong Medicine and Health Key Laboratory of Laboratory Medicine, Jinan, Shandong, China.
² Department of Pathogen Biology, School of Clinical and Basic Medical Sciences, Shandong First Medical University and Shandong Academy of Medical Sciences, Jinan, Shandong, China.
³ Department of Clinical Microbiology Laboratory, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, China.
⁴ Key Laboratory of Medical Microbiology and Parasitology, Department of Microbiology, Guizhou Medical University, Guiyang, China.

Abstract

Introduction: To eradicate Helicobacter pylori (H. pylori) and reduce the risk of gastric cancer, a sensitive, specific, convenient, and simple detection method is needed. This study aimed to establish a novel loop-mediated isothermal amplification-lateral flow dipstick (LAMP-LFD) method for H. pylori detection.

Methods: LAMP primer design software was used to design primers for the conserved sites of the H. pylori ureB gene. UreB-FIP-labeled biotin was used for LAMP amplification, and FAM-labeled probes were specifically hybridized with LAMP amplification products, which were then detected by LFD. In addition, a clinical study was conducted to assess LAMP-LFD in 20 fecal samples.

Results: The results of the optimization indicated that H. pylori could be specifically detected by LFD without cross-reaction with other non-H. pylori bacteria when the LAMP was performed at 65°C for 60 min. The lower limit of the detection method was 10² copies/μL, which was 100 times the sensitivity of polymerase chain reaction (PCR). H. pylori-positive fecal samples were detected by LAMP-LFD in 13/20 patients.

Discussion: In conclusion, a new LAMP-LFD assay has been fully established and confirmed for H. pylori detection. The entire process can be completed in approximately 1.5 h, with the advantages of strong specificity, high sensitivity, and simple operation. This study provides a novel potential method for the detection of H. pylori in the clinical settings of primary hospitals and low-resource countries.

Keywords: Helicobacter pylori; lateral flow dipstick; loop-mediated isothermal amplification; novel detection method; ureB gene.

Grants and funding

This study was supported by the National Foundation of Natural Science of China (Grant No. 81602455), the Science Foundation of Shandong Province (Grant No. ZR2013HM033), and the Taiwan Science and Technology Innovation Development Project (Grant Nos. 2020NS136 and 2021NS406).