AN APPROACH FOR DETECTION OF ESCHERICHIA COLI BY THE PCR TECHNIQUE

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Aim: Escherichia coli (E. coli) is a major cause of extra intestinal infections such as neonatal meningitis, pyelonephritis, cystitis, prostatitis and chorioamnionitis. Culture is the gold standard for detection of infections but it is frequently insufficient in clinical management because it sometimes fails to detect E coli (because of the existence of inhibitory substances, small sample volumes and/or killing of bacteria by antibiotics) and is also time-consuming. In this study, in an attempt to provide an alternative we tested the polymerase chain reaction (PCR) for detection of E. coli. Methods: Two common E. coli strains and twenty E. coli samples obtained from patients in Nagoya City University Hospital at concentrations of 3 X 10^8 cells/mm^3 were investigated using PCR by phylogenetic grouping and also determination of eight virulence factors (fimH, iucC, sfa/foc, papC, neuC, hly, afa and ibeA ). Results: In the ATCC 25922 and 35218 strains of the phylogenic grouping, bands for ChuA, YjaA TspE4.C2 were shown, allowing classification as group B2. With virulence genes, ATCC 25922 showed five bands (fimH, sfa/foc, iucC, papC, and ibeA) and ATCC 35218 showed four (sfa/foc, iucC, papC, and ibeA). Of the twenty E. coli samples, 10 were classified as group B2, 9 as group D and one as group A. Of the virulence factors, iucC was present in 19/20, fimH and saf/foc in 11/20, ibe A in 4/20, papC and hly in 3/20, afa in 2/20 and neuC in 1/20. Conclusion : Both phylogenetic grouping and detection of virulence factors by PCR can be applied for confirmation of the presence of E. coli, compensating for the unreliability of standard culture.
名古屋市立大学の論文

AN APPROACH FOR DETECTION OF ESCHERICHIA COLI BY THE PCR TECHNIQUE

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