Comparison of five real-time PCR assays for detecting virulence genes in isolates of Escherichia coli from septicaemic neonatal foals

S. Mapes, D. M. Rhodes, William D Wilson, C. M. Leutenegger, Nicola Pusterla

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Fifty-five isolates of Escherichia coli from septicaemic neonatal foals were used to validate five real-time PCR assays targeting different known virulence factor genes: curli fibre (csgD), ferric hydroxamate uptake (fhuA), type 1A pilin (fimA), aerobactin (lutA) and yersiniabactin (fyuA). A PCR assay targeting a universal sequence of the bacterial 16S rRNA gene served as quality control. The PCR assays showed good analytical specificity and sensitivity on the basis of sequencing the PCR products, their lack of cross-reactivity with non-E coli organisms, high amplification efficiency and a limit of detection as low as 25 E coli colony-forming units. There were differences between the detection rates and amplification efficiencies for the five virulence genes. The PCR assays targeting genes csgD, fhuA and fyuA were able to detect all 55 E coli isolates, with gene csgD having the best amplification efficiency. The lowest detection rate and amplification efficiency of the E coli isolates was found for the lutA gene.

Original languageEnglish (US)
Pages (from-to)716-718
Number of pages3
JournalVeterinary Record
Volume161
Issue number21
StatePublished - Nov 24 2007

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foals
Virulence
Real-Time Polymerase Chain Reaction
quantitative polymerase chain reaction
virulence
Escherichia coli
Polymerase Chain Reaction
assays
Genes
genes
Fimbriae Proteins
detection limit
analytical specificity
Gene Targeting
Virulence Factors
uptake mechanisms
rRNA Genes
Quality Control
Limit of Detection
gene targeting

ASJC Scopus subject areas

  • veterinary(all)

Cite this

Comparison of five real-time PCR assays for detecting virulence genes in isolates of Escherichia coli from septicaemic neonatal foals. / Mapes, S.; Rhodes, D. M.; Wilson, William D; Leutenegger, C. M.; Pusterla, Nicola.

In: Veterinary Record, Vol. 161, No. 21, 24.11.2007, p. 716-718.

Research output: Contribution to journalArticle

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