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Journal of Clinical Microbiology, April 2004, p. 1773-1776, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1773-1776.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Detection of the Escherichia coli Group 2 Polysaccharide Capsule Synthesis Gene kpsM by a Rapid and Specific PCR-Based Assay

James R. Johnson1,2* and Timothy T. O'Bryan1,2

Mucosal and Vaccine Research Center, Minneapolis VA Medical Center,1 Department of Medicine, University of Minnesota, Minneapolis, Minnesota2

Received 26 September 2003/ Returned for modification 15 December 2003/ Accepted 18 December 2003

A rapid and simple PCR-based assay for detection of the group 2 capsule synthesis gene kpsM of Escherichia coli was designed and validated. When combined with the published group 2 primers (kpsIIf, 5'-GCGCATTTGCTGATACTGTTG-3'; kpsIIr, 5'-CATCCAGACGATAAGCATGAGCA-3'), the new primers (the kpsIIf primer and a new reverse primer K2r, 5'-AGGTAGTTCAGACTCACACCT-3') allowed specific identification by exclusion of the heretofore elusive K2 kpsM variant. The primers yielded the predicted amplicon when multiplexed with other primers and used under varied assay conditions, including a range of concentrations of individual reaction mixture ingredients and of annealing temperatures (from 54 to 64°C).

* Corresponding author. Mailing address: Infectious Diseases (111F), Minneapolis VA Medical Center, 1 Veterans Dr., Minneapolis, MN 55417. Phone: (612) 467-4185. Fax: (612) 727-5995. E-mail: johns007{at}umn.edu.

Journal of Clinical Microbiology, April 2004, p. 1773-1776, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1773-1776.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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