Establishment of loop‑mediated isothermal amplification for rapid detection of Pseudomonas aeruginosa

Li,Chao; Shi,Yaoqiang; Yang,Guangying; Xia,Xue‑Shan; Mao,Xiaoqin; Fang,Yue; Zhang,A‑Mei; Song,Yuzhu

doi:10.3892/etm.2018.6910

January-2019 Volume 17 Issue 1

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January-2019 Volume 17 Issue 1

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Article Open Access

Establishment of loop‑mediated isothermal amplification for rapid detection of Pseudomonas aeruginosa

Authors:
- Chao Li
- Yaoqiang Shi
- Guangying Yang
- Xue‑Shan Xia
- Xiaoqin Mao
- Yue Fang
- A‑Mei Zhang
- Yuzhu Song
View Affiliations / Copyright

Affiliations: Molecular Diagnosis Laboratory, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, Yunnan 650500, P.R. China, Molecular Medicine Center of Yunnan Province, Kunming, Yunnan 650500, P.R. China

Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
Pages: 131-136
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Published online on: October 31, 2018

https://doi.org/10.3892/etm.2018.6910
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Abstract

Pseudomonas aeruginosa is one of the three most pathogenic bacteria that frequently cause life‑threatening opportunistic human infections, pneumonia, and lower respiratory tract infections in immunocompromised hosts, particularly in the burns ward. The present study aimed to establish a loop‑mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of P. aeruginosa‑specific gene hypothetical protein (GenBank ID: 882161). The gene was obtained through local and online BLAST, and specific primers were designed for this gene. Reaction conditions were optimized at 65˚C for 30 min and 80˚C for 2 min, whereas the reaction system contained 5.2 mM Mg2+, 8 U Bst 2.0 DNA polymerase, 1.4 mM deoxyribonucleotide and 0.2 and 1.6 µM of the outer and inner primers, respectively. The LAMP method was evaluated using 150 P. aeruginosa and 170 non‑P. aeruginosa strains. Positive reactions were observed on 150 P. aeruginosa strains, whereas all non‑P. aeruginosa strains exhibited negative results. Plasmids with the specific gene and mouse blood with P. aeruginosa were used for sensitivity assay. The detection limit of LAMP was 1 bacterium/reaction. Results indicated that the LAMP method targeted to hypothetical protein is a fast, specific, sensitive, inexpensive and suitable method for detection of P. aeruginosa.

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Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Establishment of loop‑mediated isothermal amplification for rapid detection of Pseudomonas aeruginosa

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