This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Al-Samarrai, T. H.
Right arrow Articles by Schmid, J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Al-Samarrai, T. H.
Right arrow Articles by Schmid, J.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, December 2000, p. 4445-4452, Vol. 38, No. 12
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Simple and Inexpensive but Highly Discriminating Method for Computer-Assisted DNA Fingerprinting of Pseudomonas aeruginosa

Taha H. Al-Samarrai,1 Ningxin Zhang,1 Iain L. Lamont,2 Lois Martin,2 John Kolbe,3 Margaret Wilsher,3 Arthur J. Morris,4 and Jan Schmid1,*

Institute of Molecular BioSciences, Massey University, Palmerston North,1 Department of Biochemistry, University of Otago, Dunedin,2 and Departments of Respiratory Services3 and Clinical Microbiology,4 Green Lane Hospital, Auckland, New Zealand

Received 16 May 2000/Returned for modification 18 August 2000/Accepted 18 September 2000

We describe here a method for computer-assisted fingerprinting of Pseudomonas aeruginosa. In this method, DNA is digested with SalI, and bands with molecular sizes of >= 9.7 kb are visually scored after electrophoresis on agarose gels. Pattern scores are entered into a Microsoft Excel database. In scoring, the number of bands within each of a set of molecular size ranges is scored, rather than the absolute molecular size of each band, substantially enhancing the speed and reproducibility of the method, while eliminating the need for using expensive gel scanning equipment and software. Pattern scores are used to generate matrices of genetic distance values, which can be visualized in neighbor-joining trees. The method reliably distinguishes two epidemiologically unrelated isolates in 99.3% of all comparisons. The genetic relationships between isolates observed with the method were consistent with those obtained by analysis of two P. aeruginosa genes, indicating that it provides valid estimates of genetic divergence between isolates. Using the method, respiratory tract isolates from cystic fibrosis patients in Green Lane Hospital in Auckland, New Zealand, were shown to be genetically less diverse than epidemiologically unrelated isolates from other patients. This finding was not due to the existence of clusters of related strains specialized toward colonization of the respiratory tract and thus was indicative of transmission between patients. Analysis of multiple isolates from individual cystic fibrosis patients suggested that up to five separate clusters of genetically related strains may simultaneously be present in a patient. The method described should significantly enhance our ability to investigate the epidemiology of P. aeruginosa.

* Corresponding author. Mailing address: Institute of Molecular BioSciences, Massey University, Private Bag 11222, Palmerston North, New Zealand. Phone: 64-6-350-4018. Fax: 64-6-350-5688. E-mail: J.Schmid{at}massey.ac.nz.

Journal of Clinical Microbiology, December 2000, p. 4445-4452, Vol. 38, No. 12
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Schmid, J., Ling, L. J., Leung, J. L. S., Zhang, N., Kolbe, J., Wesley, A. W., Mills, G. D., Brown, P. J., Jones, D. T., Laing, R. T. R., Pattemore, P. K., Taylor, D. R., Grimwood, K. (2008). Pseudomonas aeruginosa transmission is infrequent in New Zealand cystic fibrosis clinics. Eur Respir J 32: 1583-1590 [Abstract] [Full Text]  
  • Jones, W. T., Al-Samarrai, T., Reeves, J. M., Ryan, G. B., Kirk, C. A., Vincze, E., Harvey, D., McCambridge, M., Greenwood, D., Reynolds, P. H. S. (2004). The trans-Acting Protein Interacting with the DNA Motif Proximal to the Transcriptional Start Site of Plant L-Asparaginase Is Bacterial Sarcosine Oxidase. J. Bacteriol. 186: 811-817 [Abstract] [Full Text]  
  • de Chial, M., Ghysels, B., Beatson, S. A., Geoffroy, V., Meyer, J. M., Pattery, T., Baysse, C., Chablain, P., Parsons, Y. N., Winstanley, C., Cordwell, S. J., Cornelis, P. (2003). Identification of type II and type III pyoverdine receptors from Pseudomonas aeruginosa. Microbiology 149: 821-831 [Abstract] [Full Text]  
  • Norman, R. S., Frontera-Suau, R., Morris, P. J. (2002). Variability in Pseudomonas aeruginosa Lipopolysaccharide Expression during Crude Oil Degradation. Appl. Environ. Microbiol. 68: 5096-5103 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»