Abstract
We developed and evaluated the specificity and sensitivity of a simple loop-mediated isothermal amplification (LAMP) method for rapid detection of P. aeruginosa strains. The optimal reaction condition was found to be 65°C for 45 min, with the detection limit as 100 fg DNA/tube and 10 CFU/reaction. Application of LAMP assays were performed 426 clinical samples (including 252 P. aeruginosa and 174 non- P. aeruginosa isolates) using a rapid procedure and easy result confirmation. Sensitivity of LAMP and PCR assays was found to be 97.6% (246/252) and 90.5% (228/252), respectively; with a 100% specificity for both assays.
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References
Deschaght P, de Baere T, van Simaey L, van Daele S, de Baets F, de Vos D, Pirnay JP, Vaneechoutte M (2009) Comparison of the sensitivity of culture, PCR and quantitative real-time PCR for the detection of Pseudomonas aeruginosa in sputum of cystic fibrosis patients. BMC Microbiol 9:244
Jarvis WR, Martone WJ (1992) Predominant pathogens in hospital infections. J Antimicrob Chemother 29(Suppl A):19–24
Lodeng AGG, Ahlen C, Lysvand H, Mandal LH, Iversen OJ (2006) Development and evaluation of a new PCR assay for detection of Pseudomonas aeruginosa D genotype. Clin Microbiol Infect 12:761–768
Schwartz T, Volkmann H, Kirchen S, Kohnen W, Schon-Holz K, Jansen B, Obst U (2006) Real-time PCR detection of Pseudomonas aeruginosa in clinical and municipal wastewater and genotyping of the ciprofloxacin-resistant isolates. FEMS Microbiol Ecol 57:158–167
Spencer RC (1996) Predominant pathogens found in the European prevalence of infection in intensive care study. Eur J Clin Microbiol Infect Dis 15:281–285
Xu Z, Shi L, Zhang C, Zhang L, Li X, Cao Y, Li L, Yamasaki S (2007) Nosocomial infection caused by class 1 integron-carrying Staphylococcus aureus in a hospital in South China. Clin Microbiol Infect 13:980–984
Xu Z, Li L, Alam MJ, Zhang L, Yamasaki S, Shi L (2008a) First confirmation of integron-bearing methicillin-resistant Staphylococcus aureus. Curr Microbiol 57:264–268
Xu Z, Shi L, Alam MJ, Li L, Yamasaki S (2008b) Integron-bearing methicillin-resistant coagulase-negative staphylococci in South China, 2001–2004. FEMS Microbiol Lett 278:223–230
Xu Z, Li L, Shirtliff ME, Alam MJ, Yamasaki S, Shi L (2009) Occurrence and characteristics of class 1 and 2 integrons in Pseudomonas aeruginosa isolates from patients in Southern China. J Clin Microbiol 47:230–234
Zhao X, Li Y, Wang L, You L, Xu Z, Li L, He X, Liu Y, Wang J, Yang L (2009) Development and application of a loop-mediated isothermal amplification method on rapid detection Escherichia coli O157 strains from food samples. Mol Biol Rep. doi: 10.1007/s11033-009-9700-6
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This work was supported by Science Foundation of Ministry of Education of China (706046), National Natural Science Foundation of China (20436020) and State Scholarship Fund of China Scholarship Council (2008615044).
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Xihong Zhao and Li Wang contributed equally to this work.
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Zhao, X., Wang, L., Li, Y. et al. Development and application of a loop-mediated isothermal amplification method on rapid detection of Pseudomonas aeruginosa strains. World J Microbiol Biotechnol 27, 181–184 (2011). https://doi.org/10.1007/s11274-010-0429-0
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DOI: https://doi.org/10.1007/s11274-010-0429-0