Peer-Reviewed Journal Details
Mandatory Fields
Minogue, E,Reddington, K,Dorai-Raj, S,Tuite, N,Clancy, E,Barry, T
2013
September
Journal Of Industrial Microbiology & Biotechnology
Diagnostics method for the rapid quantitative detection and identification of low-level contamination of high-purity water with pathogenic bacteria
Published
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Optional Fields
Burkholderia genus Pseudomonas aeruginosa Spiked high-purity water samples REAL-TIME PCR BURKHOLDERIA-CEPACIA COMPLEX TO-PERSON TRANSMISSION PSEUDOMONAS-AERUGINOSA MICROBIAL-CONTAMINATION LISTERIA-MONOCYTOGENES RNASE-P NOSOCOMIAL INFECTIONS PURIFICATION SYSTEM ENDOSCOPE WASHER
40
1005
1013
High-purity water (HPW) can be contaminated with pathogenic microorganisms, which may result in human infection. Current culture-based techniques for the detection of microorganisms from HPW can be slow and laborious. The aim of this study was to develop a rapid method for the quantitative detection and identification of pathogenic bacteria causing low-level contamination of HPW. A novel internally controlled multiplex real-time PCR diagnostics assay was designed and optimized to specifically detect and identify Pseudomonas aeruginosa and the Burkholderia genus. Sterile HPW, spiked with a bacterial load ranging from 10 to 10(3) cfu/100 ml, was filtered and the bacterial cells were removed from the filters by sonication. Total genomic DNA was then purified from these bacteria and subjected to testing with the developed novel multiplex real-time PCR diagnostics assay. The specific P. aeruginosa and Burkholderia genus assays have an analytical sensitivity of 3.5 genome equivalents (GE) and 3.7 GE, respectively. This analysis demonstrated that it was possible to detect a spiked bacterial load of 1.06 x 10(2) cfu/100 ml for P. aeruginosa and 2.66 x 10(2) cfu/100 ml for B. cepacia from a 200-ml filtered HPW sample. The rapid diagnostics method described can reliably detect, identify, and quantify low-level contamination of HPW with P. aeruginosa and the Burkholderia genus in < 4 h. We propose that this rapid diagnostics method could be applied to the pharmaceutical and clinical sectors to assure the safety and quality of HPW, medical devices, and patient-care equipment.
DOI 10.1007/s10295-013-1295-1
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