Frontiers in Cellular and Infection Microbiology (Jan 2023)
Development of a novel loop-mediated isothermal amplification assay for ß-lactamase gene identification using clinical isolates of Gram-negative bacteria
- Eun Jin Kim,
- Eun Jin Kim,
- Eun Jin Kim,
- Jiwon Lee,
- Jiwon Lee,
- Jiwon Lee,
- Youngbae Yoon,
- Youngbae Yoon,
- Youngbae Yoon,
- Donghyun Lee,
- Donghyun Lee,
- Donghyun Lee,
- Yeongjun Baek,
- Yeongjun Baek,
- Yeongjun Baek,
- Chika Takano,
- Jun Sakai,
- Takahiro Iijima,
- Dai Kanamori,
- Humphrey Gardner,
- Robert E. McLaughlin,
- Paul E. Kilgore,
- Akihiro Nakamura,
- Takashi Ogihara,
- Satoshi Hayakawa,
- Tomonori Hoshino,
- Dong Wook Kim,
- Dong Wook Kim,
- Mitsuko Seki,
- Mitsuko Seki
Affiliations
- Eun Jin Kim
- Division of Pediatric Dentistry, Department of Human Development and Fostering, Meikai University School of Dentistry, Saitama, Japan
- Eun Jin Kim
- Department of Pharmacy, College of Pharmacy, Hanyang University, Ansan, Republic of Korea
- Eun Jin Kim
- Institute of Pharmacological Research, Hanyang University, Ansan, Republic of Korea
- Jiwon Lee
- Division of Pediatric Dentistry, Department of Human Development and Fostering, Meikai University School of Dentistry, Saitama, Japan
- Jiwon Lee
- Department of Pharmacy, College of Pharmacy, Hanyang University, Ansan, Republic of Korea
- Jiwon Lee
- Institute of Pharmacological Research, Hanyang University, Ansan, Republic of Korea
- Youngbae Yoon
- Division of Pediatric Dentistry, Department of Human Development and Fostering, Meikai University School of Dentistry, Saitama, Japan
- Youngbae Yoon
- Department of Pharmacy, College of Pharmacy, Hanyang University, Ansan, Republic of Korea
- Youngbae Yoon
- Institute of Pharmacological Research, Hanyang University, Ansan, Republic of Korea
- Donghyun Lee
- Division of Pediatric Dentistry, Department of Human Development and Fostering, Meikai University School of Dentistry, Saitama, Japan
- Donghyun Lee
- Department of Pharmacy, College of Pharmacy, Hanyang University, Ansan, Republic of Korea
- Donghyun Lee
- Institute of Pharmacological Research, Hanyang University, Ansan, Republic of Korea
- Yeongjun Baek
- Division of Pediatric Dentistry, Department of Human Development and Fostering, Meikai University School of Dentistry, Saitama, Japan
- Yeongjun Baek
- Department of Pharmacy, College of Pharmacy, Hanyang University, Ansan, Republic of Korea
- Yeongjun Baek
- Institute of Pharmacological Research, Hanyang University, Ansan, Republic of Korea
- Chika Takano
- Division of Microbiology, Department of Pathology and Microbiology, Nihon University School of Medicine, Tokyo, Japan
- Jun Sakai
- Department of Infectious Disease and Infection Control, Saitama Medical University, Saitama, Japan
- Takahiro Iijima
- Division of Pediatric Dentistry, Department of Human Development and Fostering, Meikai University School of Dentistry, Saitama, Japan
- Dai Kanamori
- Division of Pediatric Dentistry, Department of Human Development and Fostering, Meikai University School of Dentistry, Saitama, Japan
- Humphrey Gardner
- Harbour Biomed, Natick, MA, United States
- Robert E. McLaughlin
- Institute for Life Science Entrepreneurship, Union, NJ, United States
- Paul E. Kilgore
- Department of Pharmacy Practice, Eugene Applebaum College of Pharmacy & Health Sciences, Wayne State University, Detroit, MI, United States
- Akihiro Nakamura
- Division of Pediatric Dentistry, Department of Human Development and Fostering, Meikai University School of Dentistry, Saitama, Japan
- Takashi Ogihara
- Division of Pediatric Dentistry, Department of Human Development and Fostering, Meikai University School of Dentistry, Saitama, Japan
- Satoshi Hayakawa
- Division of Microbiology, Department of Pathology and Microbiology, Nihon University School of Medicine, Tokyo, Japan
- Tomonori Hoshino
- Division of Pediatric Dentistry, Department of Human Development and Fostering, Meikai University School of Dentistry, Saitama, Japan
- Dong Wook Kim
- Department of Pharmacy, College of Pharmacy, Hanyang University, Ansan, Republic of Korea
- Dong Wook Kim
- Institute of Pharmacological Research, Hanyang University, Ansan, Republic of Korea
- Mitsuko Seki
- Division of Pediatric Dentistry, Department of Human Development and Fostering, Meikai University School of Dentistry, Saitama, Japan
- Mitsuko Seki
- Division of Microbiology, Department of Pathology and Microbiology, Nihon University School of Medicine, Tokyo, Japan
- DOI
- https://doi.org/10.3389/fcimb.2022.1000445
- Journal volume & issue
-
Vol. 12
Abstract
Rapid evaluation of antimicrobial susceptibility is important in the treatment of nosocomial infections by Gram-negative bacteria, which increasingly carry carbapenemases and metallo-β-lactamases. We developed loop-mediated isothermal amplification (LAMP)-based assays for four β-lactamase genes (blaKPC, blaNDM-1, blaIMP-1 group, and blaVIM). The assays were evaluated using eight reference bacterial strains (Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, and Acinetobacter bereziniae) harboring six β-lactamase genes. A total of 55 Gram-negative bacterial strains, including 47 clinical P. aeruginosa isolates, fully characterized by next-generation sequencing (NGS), were used to evaluate the LAMP assays. The results were compared to those of conventional PCR. The LAMP assays were able to detect as few as 10 to 100 copies of a gene, compared to 10 to 104 copies for conventional PCR. The LAMP assay detected four β-lactamase genes with a sensitivity similar to that using purified DNA as the template in DNA-spiked urine, sputum, and blood specimens. By contrast, the sensitivity of PCR was 1- to 100-fold lower with DNA-spiked clinical specimens. Therefore, the LAMP assays were proved to be an appropriate tool for the detection of four β-lactamases.
Keywords
- loop-mediated isothermal amplification
- ß-lactamase gene
- Gram-negative bacteria
- blaKPC
- blaNDM-1
- blaIMP-1 group
