Rapid genotyping of Pseudomonas aeruginosa using repetitive element based PCR assays

O'Carroll, M. R., Coulter, C., Bell, S. C., Syrmis, M., Wainwright, C. E., Nissen, M. and Sloots, T. (2003). Rapid genotyping of Pseudomonas aeruginosa using repetitive element based PCR assays. In: Journal of Cystic Fibrosis: Proceedings of the 26th European CF Conference. 26th European CF Conference, Belfast, Ireland, (S38-S38). 5–7 June , 2003. doi:10.1016/S1569-1993(03)00027-4


Author O'Carroll, M. R.
Coulter, C.
Bell, S. C.
Syrmis, M.
Wainwright, C. E.
Nissen, M.
Sloots, T.
Title of paper Rapid genotyping of Pseudomonas aeruginosa using repetitive element based PCR assays
Conference name 26th European CF Conference
Conference location Belfast, Ireland
Conference dates 5–7 June , 2003
Proceedings title Journal of Cystic Fibrosis: Proceedings of the 26th European CF Conference   Check publisher's open access policy
Place of Publication Netherlands
Publisher Elsevier Science
Publication Year 2003
DOI 10.1016/S1569-1993(03)00027-4
ISSN 1569-1993
Volume 2
Issue 1 Supplement
Start page S38
End page S38
Total pages 1
Language eng
Formatted Abstract/Summary
Background:
Transmissible strains of P. aeruginosa have recently been reported in several adult and paediatric cystic fibrosis (CF) clinics. Pulse field gel electrophoresis (PFGE) is currently recognised as the gold standard of molecular typing techniques for P. aeruginosa, however, it is labour intensive and limited by high cost and extended turnaround times. Transmissible strains represent an expanding problem and routine microbiological surveillance may be necessary in the near future. A screening method that is rapid, relatively inexpensive and highly specific is required. We compared two repetitive element based PCR genotyping assays with PFGE.

Methods:
Sputum was collected prospectively from all productive patients attending the CF clinics at the RCH and TPCH and in Brisbane from December 2001. Genomic DNA was extracted from P. aeruginosa isolates using an AquaPure Genomic DNA kit (BioRAD). Two Rep PCR assays (Enterobacterial  epetitive Intergenic Consensus (ERIC) PCR and BOX PCR) were modifications of  reviously published methods. PFGE was performed following digestion with SpeI, results compared with Gelcompar and Tenover criteria were applied to determine relatedness.

Results:

One hundred and sixty three isolates from the first 100 patients (50 from each clinic) were tested using both techniques. Each of the isolates that had two or more band differences by either the BOX or ERIC PCR assay was unrelated by the PFGE method. These results were concordant without exception.

Conclusions:

We have developed a rapid PCR based genotyping method with high pecificity. This could serve as a screening tool for microbiological surveillance inCF clinics. Whilst this method does not eliminate the need for PFGE, it will significantly reduce the number of isolates that need testing by this method, thereby reducing costs and improving efficiency. Supported by The Royal Children’s Hospital Foundation.
Subjects CX
321027 Respiratory Diseases
730110 Respiratory system and diseases (incl. asthma)
Q-Index Code CX
Additional Notes Erratum to “Abstract Book—26th European CF Conference, Belfast, June 5–7, 2003 Journal of Cystic Fibrosis 2S1 (2004) 3–112” Journal of Cystic Fibrosis, Volume 3, Issue 2, June 2004, Page 139

 
Versions
Version Filter Type
Citation counts: Google Scholar Search Google Scholar
Created: Wed, 15 Aug 2007, 01:41:28 EST