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

  • Published online on: October 31, 2018     https://doi.org/10.3892/etm.2018.6910
  • Pages: 131-136
  • Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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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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January-2019
Volume 17 Issue 1

Print ISSN: 1792-0981
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Spandidos Publications style
Li C, Shi Y, Yang G, Xia XS, Mao X, Fang Y, Zhang AM and Song Y: Establishment of loop‑mediated isothermal amplification for rapid detection of Pseudomonas aeruginosa. Exp Ther Med 17: 131-136, 2019
APA
Li, C., Shi, Y., Yang, G., Xia, X., Mao, X., Fang, Y. ... Song, Y. (2019). Establishment of loop‑mediated isothermal amplification for rapid detection of Pseudomonas aeruginosa. Experimental and Therapeutic Medicine, 17, 131-136. https://doi.org/10.3892/etm.2018.6910
MLA
Li, C., Shi, Y., Yang, G., Xia, X., Mao, X., Fang, Y., Zhang, A., Song, Y."Establishment of loop‑mediated isothermal amplification for rapid detection of Pseudomonas aeruginosa". Experimental and Therapeutic Medicine 17.1 (2019): 131-136.
Chicago
Li, C., Shi, Y., Yang, G., Xia, X., Mao, X., Fang, Y., Zhang, A., Song, Y."Establishment of loop‑mediated isothermal amplification for rapid detection of Pseudomonas aeruginosa". Experimental and Therapeutic Medicine 17, no. 1 (2019): 131-136. https://doi.org/10.3892/etm.2018.6910