Detection of the Escherichia coli group 2 polysaccharide capsule synthesis Gene kpsM by a rapid and specific PCR-based assay.
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 degrees C).
Johnson
James R
JR
Mucosal and Vaccine Research Center, Minneapolis VA Medical Center, Minnesota. johns007@umn.edu
O'Bryan
Timothy T
TT
D016667:Bacterial Capsules / Q000378:metabolism; D017931:DNA Primers; D004926:Escherichia coli / Q000145:classification* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics*; D005838:Genotype; D006801:Humans; D026901:Membrane Transport Proteins / Q000235:genetics*; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity; D013997:Time Factors
D017931:DNA Primers; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; C068104:kpsM protein, E coli