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World Journal of Microbiology and Biotechnology

, Volume 27, Issue 1, pp 181–184 | Cite as

Development and application of a loop-mediated isothermal amplification method on rapid detection of Pseudomonas aeruginosa strains

  • Xihong Zhao
  • Li Wang
  • Yanmei Li
  • Zhenbo XuEmail author
  • Lin Li
  • Xiaowei HeEmail author
  • Yao Liu
  • Jihua Wang
  • Liansheng Yang
Short Communication

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.

Keywords

Loop-mediated isothermal amplification (LAMP) P. aeruginosa Rapid detection 

Notes

Acknowledgments

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).

References

  1. 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 9:244CrossRefGoogle Scholar
  2. Jarvis WR, Martone WJ (1992) Predominant pathogens in hospital infections. J Antimicrob Chemother 29(Suppl A):19–24Google Scholar
  3. 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 12:761–768CrossRefGoogle Scholar
  4. 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 57:158–167CrossRefGoogle Scholar
  5. Spencer RC (1996) Predominant pathogens found in the European prevalence of infection in intensive care study. Eur J Clin Microbiol Infect Dis 15:281–285CrossRefGoogle Scholar
  6. 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 13:980–984CrossRefGoogle Scholar
  7. Xu Z, Li L, Alam MJ, Zhang L, Yamasaki S, Shi L (2008a) First confirmation of integron-bearing methicillin-resistant Staphylococcus aureus. Curr Microbiol 57:264–268CrossRefGoogle Scholar
  8. 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 278:223–230CrossRefGoogle Scholar
  9. 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 47:230–234CrossRefGoogle Scholar
  10. 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

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Xihong Zhao
    • 1
  • Li Wang
    • 2
  • Yanmei Li
    • 3
  • Zhenbo Xu
    • 1
    • 4
    Email author
  • Lin Li
    • 1
  • Xiaowei He
    • 1
    Email author
  • Yao Liu
    • 5
  • Jihua Wang
    • 6
  • Liansheng Yang
    • 1
  1. 1.College of Light Industry and Food SciencesSouth China University of TechnologyGuangzhouChina
  2. 2.Food Safety Key Laboratory of Guangdong Province, College of Food ScienceSouth China Agricultural UniversityGuangzhouChina
  3. 3.Guangzhou Medical CollegeGuangzhouChina
  4. 4.Department of Microbial Pathogenesis, Dental SchoolUniversity of Maryland-BaltimoreBaltimore, MarylandUSA
  5. 5.Zhongshan Supervision Testing Institute of Quality & MetrologyZhongshanChina
  6. 6.Guangzhou Wondfo Biotechnology CompanyGuangzhouChina

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