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Comparison of five real-time PCR assays for detecting virulence genes in isolates of Escherichia coli from septicaemic neonatal foals.
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.
Publication Date: 2007-11-27 PubMed ID: 18037693DOI: 10.1136/vr.161.21.716Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This research investigates the efficiency of five different real-time PCR assays in detecting virulent genes in Escherichia coli samples collected from sick newborn foals. The results indicate varying efficiency levels and detection capabilities among the assays, with the csgD gene displaying the best performance.
Research Methodology
- The study involved 55 Escherichia coli isolates obtained from septic newborn foals.
- Five distinct real-time PCR assays were employed, each targeting a different known virulence factor gene. These included csgD (curli fibre), fhuA (ferric hydroxamate uptake), fimA (type 1A pilin), lutA (aerobactin), and fyuA (yersiniabactin).
- A PCR assay specifically targeting the universal bacterial 16S rrna gene was employed as a quality control measure.
Results and Findings
- The five PCR assays demonstrated good analytical specificity and sensitivity. This conclusion was based on several factors, such as the sequencing of PCR products, the absence of cross-reactivity with non-E coli organisms, high amplification efficiency, and a detection limit as low as 25 E coli colony-forming units.
- There were disparities in terms of detection rates and amplification efficiencies across the five virulence genes. The PCR assays targeting the csgD, fhuA, and fyuA genes successfully detected all 55 E coli isolates, with the csgD gene delivering the best amplification efficiency.
- The lutA gene was, unfortunately, associated with the lowest detection rate and amplification efficiency among the E coli isolates.
Conclusion
- The findings indicate different efficiencies and detection capabilities of the various real-time PCR assays under study. This highlights the need to carefully choose the right PCR assay for detecting specific virulence genes.
- The csgD gene performed best among all in terms of amplification efficiency. This means that it can be reliably used for detecting the presence of E coli in samples.
- On the other hand, the lutA gene showed the lowest detection rate and amplification efficiency, suggesting it might not be as reliable for E coli detection. Additional research or techniques might be required to accurately detect this particular gene.
Cite This Article
APA
Mapes S, Rhodes DM, Wilson WD, Leutenegger CM, Pusterla N.
(2007).
Comparison of five real-time PCR assays for detecting virulence genes in isolates of Escherichia coli from septicaemic neonatal foals.
Vet Rec, 161(21), 716-718.
https://doi.org/10.1136/vr.161.21.716 Publication
Researcher Affiliations
- Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, 1 Shields Avenue, Davis, CA 95616, USA.
MeSH Terms
- Animals
- Animals, Newborn
- Escherichia coli / genetics
- Escherichia coli / pathogenicity
- Escherichia coli Infections / microbiology
- Escherichia coli Infections / veterinary
- Horse Diseases / microbiology
- Horses
- Polymerase Chain Reaction / methods
- Polymerase Chain Reaction / veterinary
- Sensitivity and Specificity
- Sepsis / microbiology
- Sepsis / veterinary
- Virulence / genetics
Citations
This article has been cited 13 times.- Stummer M, Frisch V, Glitz F, Hinney B, Spergser J, Krücken J, Diekmann I, Dimmel K, Riedel C, Cavalleri JV, Rümenapf T, Joachim A, Lyrakis M, Auer A. Presence of Equine and Bovine Coronaviruses, Endoparasites, and Bacteria in Fecal Samples of Horses with Colic. Pathogens 2023 Aug 15;12(8).
- Yeh JY. Prevalence and associated risk factors for Lawsonia intracellularis infection in farmed rabbits: A serological and molecular cross-sectional study in South Korea. Front Vet Sci 2023;10:1058113.
- Prutton JSW, Barnum S, Pusterla N. Evaluation of safety, humoral immune response and faecal shedding in horses inoculated with a modified-live bovine coronavirus vaccination. Equine Vet Educ 2020 Aug;32(Suppl 11):33-36.
- Sanz MG, Kwon S, Pusterla N, Gold JR, Bain F, Evermann J. Evaluation of equine coronavirus fecal shedding among hospitalized horses. J Vet Intern Med 2019 Mar;33(2):918-922.
- Hwang JM, Seo MJ, Yeh JY. Lawsonia intracellularis in the feces of wild rodents and stray cats captured around equine farms. BMC Vet Res 2017 Aug 11;13(1):233.
- Fielding CL, Higgins JK, Higgins JC, McIntosh S, Scott E, Giannitti F, Mete A, Pusterla N. Disease associated with equine coronavirus infection and high case fatality rate. J Vet Intern Med 2015 Jan;29(1):307-10.
- Sampieri F, Vannucci FA, Allen AL, Pusterla N, Antonopoulos AJ, Ball KR, Thompson J, Dowling PM, Hamilton DL, Gebhart CJ. Species-specificity of equine and porcine Lawsonia intracellularis isolates in laboratory animals. Can J Vet Res 2013 Oct;77(4):261-72.
- Pusterla N, Mapes S, Wademan C, White A, Ball R, Sapp K, Burns P, Ormond C, Butterworth K, Bartol J, Magdesian KG. Emerging outbreaks associated with equine coronavirus in adult horses. Vet Microbiol 2013 Feb 22;162(1):228-31.
- Vannucci FA, Pusterla N, Mapes SM, Gebhart C. Evidence of host adaptation in Lawsonia intracellularis infections. Vet Res 2012 Jun 20;43(1):53.
- Ayres JS, Trinidad NJ, Vance RE. Lethal inflammasome activation by a multidrug-resistant pathobiont upon antibiotic disruption of the microbiota. Nat Med 2012 May;18(5):799-806.
- Veir JK, Lappin MR. Molecular diagnostic assays for infectious diseases in cats. Vet Clin North Am Small Anim Pract 2010 Nov;40(6):1189-200.
- Mahale RP, K A, Princy A, Maheshwarappa YD, Sumana MN. Comparative evaluation of biofilm-forming capacity in uropathogenic and commensal Escherichia coli. Front Cell Infect Microbiol 2025;15:1570422.
- Li L, Li S, Ma H, Akhtar MF, Tan Y, Wang T, Liu W, Khan A, Khan MZ, Wang C. An Overview of Infectious and Non-Infectious Causes of Pregnancy Losses in Equine. Animals (Basel) 2024 Jul 2;14(13).
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