Detection of Salmonella spp in fecal specimens by use of real-time polymerase chain reaction assay.
Abstract: To use real-time polymerase chain reaction (PCR) technology to develop a highly sensitive and specific diagnostic assay for the detection of Salmonella spp in fecal specimens. Methods: 299 fecal specimens from cattle, horses, and dogs. Methods: Enrichment of fecal specimens was followed by genomic DNA extraction by use of commercially available isolation kits. Real-time PCR assay was performed to target a Salmonella spp-specific DNA segment. Results of real-time PCR assay were compared with bacterial culture results to determine relative sensitivity and specificity. Results: Use of the spaQ primer-probe set resulted in a relative sensitivity of 100% and a specificity of 98.2%, compared with bacterial culture results when tested on 299 clinical fecal specimens. Conclusions: A rapid, sensitive, and specific assay for the detection of Salmonella spp from enriched clinical fecal specimens was developed. This technique would be highly valuable in clinical settings to help avoid or mitigate the complications arising from an outbreak of salmonellosis in a herd or among patients of a veterinary hospital.
Publication Date: 2002-09-13 PubMed ID: 12224858DOI: 10.2460/ajvr.2002.63.1265Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research article presents the development of a diagnostic assay using real-time polymerase chain reaction (PCR) for the detection of Salmonella in animal fecal samples. The method developed proved highly efficient and reliable, a valuable tool for preventing or mitigating Salmonella outbreaks in clinical settings.
Objectives and Methods Used
- The goal of this research was to use real-time PCR technology to create a diagnostic assay that is sensitive and specific in detecting Salmonella species in fecal specimens.
- The study used 299 fecal specimens collected from cattle, horses, and dogs.
- The team employed a procedure involving the enrichment of fecal samples, which was followed by the extraction of genomic DNA using commercially available isolation kits.
- The real-time PCR assay was performed to identify a specific DNA segment of Salmonella species.
Comparison and Analysis of Results
- The researchers compared the results of the real-time PCR assay with traditional bacterial culture results to ascertain the relative sensitivity and specificity of their method.
- The use of the spaQ primer-probe set delivered a relative sensitivity of 100% and a specificity of 98.2% compared to the bacterial culture results. This suggests that the PCR-based method is highly effective in detecting Salmonella species in the tested animal fecal samples.
Conclusions Reached
- The researchers were successful in developing a rapid, sensitive, and precise assay for detecting Salmonella spp from enriched fecal specimens collected in a clinical setting.
- Given the results, it was concluded that this technique could be incredibly valuable in veterinary hospitals and any clinical settings that have to deal with potential Salmonella outbreaks.
- Application of this method could help health practitioners avoid or better control and mitigate the complications arising from salmonellosis outbreaks in herds or among the patients of a veterinary hospital.
Cite This Article
APA
Kurowski PB, Traub-Dargatz JL, Morley PS, Gentry-Weeks CR.
(2002).
Detection of Salmonella spp in fecal specimens by use of real-time polymerase chain reaction assay.
Am J Vet Res, 63(9), 1265-1268.
https://doi.org/10.2460/ajvr.2002.63.1265 Publication
Researcher Affiliations
- Department of Microbiology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins 80523, USA.
MeSH Terms
- Animals
- Cattle
- DNA, Bacterial / analysis
- Dogs
- Feces / microbiology
- Horses / microbiology
- Polymerase Chain Reaction / methods
- Polymerase Chain Reaction / veterinary
- Salmonella / genetics
- Salmonella / isolation & purification
- Sensitivity and Specificity
Citations
This article has been cited 17 times.- Amory H, Cesarini C, De Maré L, Loublier C, Moula N, Detilleux J, Saulmont M, Garigliany MM, Lecoq L. Relationship between the Cycle Threshold Value (Ct) of a Salmonella spp. qPCR Performed on Feces and Clinical Signs and Outcome in Horses.. Microorganisms 2023 Jul 30;11(8).
- Parra-Aguirre JC, Nosach R, Fernando C, Hill JE, Harding JCS. Improving the consistency of experimental swine dysentery inoculation strategies.. Vet Res 2023 Jun 16;54(1):49.
- Bhandari D, Chen FC, Bridgman RC. Magnetic Nanoparticles Enhanced Surface Plasmon Resonance Biosensor for Rapid Detection of Salmonella Typhimurium in Romaine Lettuce.. Sensors (Basel) 2022 Jan 9;22(2).
- Arnold CE, Pilla R, Chaffin MK, Leatherwood JL, Wickersham TA, Callaway TR, Lawhon SD, Lidbury JA, Steiner JM, Suchodolski JS. The effects of signalment, diet, geographic location, season, and colitis associated with antimicrobial use or Salmonella infection on the fecal microbiome of horses.. J Vet Intern Med 2021 Sep;35(5):2437-2448.
- Hertzer JN, Fujishiro M, Lawhon SD, Creevy KE. Treatment and management of Salmonella prostatitis in a heartworm-positive intact male dog: a case report.. BMC Vet Res 2021 Mar 30;17(1):135.
- Andruzzi MN, Krath ML, Lawhon SD, Boudreau B. Salmonella enterica subspecies houtenae as an opportunistic pathogen in a case of meningoencephalomyelitis and bacteriuria in a dog.. BMC Vet Res 2020 Nov 11;16(1):437.
- Blake AB, Cigarroa A, Klein HL, Khattab MR, Keating T, Van De Coevering P, Lidbury JA, Steiner JM, Suchodolski JS. Developmental stages in microbiota, bile acids, and clostridial species in healthy puppies.. J Vet Intern Med 2020 Nov;34(6):2345-2356.
- Villamil C, Calderon MN, Arias MM, Leguizamon JE. Validation of Droplet Digital Polymerase Chain Reaction for Salmonella spp. Quantification.. Front Microbiol 2020;11:1512.
- Arnold CE, Isaiah A, Pilla R, Lidbury J, Coverdale JS, Callaway TR, Lawhon SD, Steiner J, Suchodolski JS. The cecal and fecal microbiomes and metabolomes of horses before and after metronidazole administration.. PLoS One 2020;15(5):e0232905.
- Krath ML, Little SV, Hillhouse AE, Lawhon SD. Salmonella enterica subsp. arizonae Isolated from a Canine Clinical Case of Prostatitis.. Microbiol Resour Announc 2020 Mar 26;9(13).
- Hensel M, Meason-Smith C, Plumlee QD, Myers AN, Coleman MC, Lawhon S, Rodrigues Hoffmann A, Rech RR. Retrospective Analysis of Aetiological Agents Associated with Pulmonary Mycosis Secondary to Enteric Salmonellosis in Six Horses by Panfungal Polymerase Chain Reaction.. J Comp Pathol 2020 Jan;174:1-7.
- Shaw SD, Stämpfli H. Diagnosis and Treatment of Undifferentiated and Infectious Acute Diarrhea in the Adult Horse.. Vet Clin North Am Equine Pract 2018 Apr;34(1):39-53.
- Oliver-Espinosa O. Foal Diarrhea: Established and Postulated Causes, Prevention, Diagnostics, and Treatments.. Vet Clin North Am Equine Pract 2018 Apr;34(1):55-68.
- Gizzi AB, Oliveira ST, Leutenegger CM, Estrada M, Kozemjakin DA, Stedile R, Marcondes M, Biondo AW. Presence of infectious agents and co-infections in diarrheic dogs determined with a real-time polymerase chain reaction-based panel.. BMC Vet Res 2014 Jan 16;10:23.
- Malorny B, Paccassoni E, Fach P, Bunge C, Martin A, Helmuth R. Diagnostic real-time PCR for detection of Salmonella in food.. Appl Environ Microbiol 2004 Dec;70(12):7046-52.
- Song Y, Liu C, Finegold SM. Real-time PCR quantitation of clostridia in feces of autistic children.. Appl Environ Microbiol 2004 Nov;70(11):6459-65.
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