Real-time identification of Pseudomonas aeruginosa direct from clinical samples using a rapid extraction method and polymerase chain reaction (PCR)

Journal of Clinical Laboratory Analysis
R I JaffeC W Bates

Abstract

Pseudomonas aeruginosa has emerged as one of the most problematic Gram-negative nosocomial pathogens. Bacteremia caused by P. aeruginosa is clinically indistinguishable from other Gram-negative infections although the mortality rate is higher. This microorganism is also inherently resistant to common antibiotics. Standard bacterial identification and susceptibility testing is normally a 48-hour process and difficulty sometimes exists in rapidly and accurately identifying antimicrobial resistance. The Polymerase Chain Reaction (PCR) is a rapid and simple process for the amplification of target DNA sequences. However, many sample preparation methods are unsuitable for the clinical laboratory because they are not cost effective, take too long to perform, or do not provide a good template for PCR. Our goal was to provide same-day results to facilitate rapid diagnosis. In this report, we have utilized our rapid DNA extraction method to generate bacterial DNA direct from clinical samples for PCR. The lower detection level for P. aeruginosa was estimated to be 10 CFU/ml. In addition, we wanted to compare the results of a new rapid-cycle DNA thermocycler that uses continuous fluorescence monitoring with the results of standard thermocy...Continue Reading

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Citations

Mar 19, 2013·Applied Microbiology and Biotechnology·Hyeon Jin ChoiDong Suk Park
Jan 22, 2003·Environmental Microbiology·Jean-Paul PirnayPierre Cornelis
Aug 6, 2010·Annals of Clinical Microbiology and Antimicrobials·Vincent CattoirPatrick Legrand
May 27, 2008·Journal of Medical Case Reports·Sophie BaylePhilippe Brouqui
Feb 15, 2007·Environmental Microbiology·Sylvie ChevalierNicole Orange
Jan 20, 2010·Wound Repair and Regeneration : Official Publication of the Wound Healing Society [and] the European Tissue Repair Society·Trine R ThomsenPer H Nielsen
Jan 4, 2012·FEMS Immunology and Medical Microbiology·Vibeke B RudkjøbingThomas Bjarnsholt
Nov 30, 2004·The Lancet Infectious Diseases·Remco P H PetersChristina M J E Vandenbroucke-Grauls
Nov 7, 2017·Experimental and Therapeutic Medicine·Warren ChandaMintao Zhong

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