Salmonella spp.

Background

Salmonella is a gram-negative bacteria and a leading cause of enterocolitis in susceptible horses. Clinical cases range from asymptomatic to acute and severe diarrhea to death. Infection can occur via many vehicles including the environment, feed, water and contact with other animals. Further, it is possible for disease outbreaks and population epidemics to occur depending on the organism, host and degree of exposure. It is also possible for subclinical shedders to endanger the health of hospitalized patients and clinic personnel. It is thus recommended that PCR be used for rapid and specific pathogen detection to maintain biosecurity.1 Microbiological culture is typically used in tandem as a standard of care due to only small concentrations of salmonella spp. commonly being present. PCR has been shown as a more reliable method for salmonella detection than culture. However, 24-hour culture is used to amplify signal in a sample in case pathogenic loads are below an assay’s limit of detection (e.g. enrichment broth).2 This Fluxergy assay targets a conserved gene responsible for invasion of epithelial cells by all pathogenic strains of salmonella. Fluxergy further offers a 12-hour incubation buffer that can be used with the Fluxergy salmonella spp. assay to ensure absence or presence of pathogen.

Key Benefits

Simple workflow

No sample preparation

Automated data interpretation

Inhouse outbreak prevention

Comprehensive detection of salmonella species

Optional enrichment step

Sample to result in 30 minutes

Assay Specifications

Sample Type Fecal Swab
Species Equine
Storage Condition Fluxergy Card – Room temperature @ ~23° C
Fluxergy Buffers – Frozen @ -20°C

Kit Contents

  • 6” Rayon swab
  • Fluxergy salmonella spp. Buffer 1
  • Fluxergy salmonella spp. Buffer 2
  • Fluxergy Card
  • Fluxergy Enrichment Buffer (Optional)

Customer supplied reagents and supplies

  • Clinical sample for testing

Sample collection method

Fecal swab. It is recommended that the head of the swab is fully coated with sample material. For optimal performance, collect feces to be tested in a sterile 2 oz specimen cup. Fully submerge the head of the swab into the sample for 5 seconds. Place stool coated swab into the swab tube. The opposite end of the tube serves as a tight-fitting cap. Collect in sterile container. Store sample in refrigerator or at 4°C if not testing immediately after sample collection.

Intended use

The Fluxergy salmonella spp. assay is a qualitative test for the rapid detection of pathogenic salmonella spp. in equine fecal samples.

Positive PCR Result: indicates that DNA or RNA of the target organism is present in the tested sample.

Negative PCR Result: indicates that DNA or RNA of the target organism was not detected in the tested sample. However, a negative PCR results may also indicate that the number of target organisms is below the limit of detection.

Warnings and Precautions

  • Fluxergy’s salmonella spp. assay is for Research Use Only (RUO). It is not intended for diagnostic use.
  • Fluxergy’s salmonella spp. assay is compatible only with the Fluxergy Analyzer Beta device.
  • All specimens should be handled as potentially infectious agents and according to universal safety precautions.
  • This Fluxergy salmonella spp. assay is compatible only for equine fecal swabs.
  • Contamination of the sample and kit contents may lead to erroneous results. Use aseptic technique and a clean workspace.
  • Store Fluxergy assay kit at recommended storage temperature and conduct assay within specified environment (e.g. temperature and humidity) for optimal performance.
  • Follow appropriate specimen collection, storage and processing for optimal performance.
  • Use Fluxergy supplied swabs, reagents, pipettes and pipette tips to conduct assay for optimal performance.

Detection of Clinical Sample

PCR was conducted using the Fluxergy Analyzer Beta with clinical salmonella positive equine feces and a Fluxergy buffer with salmonella specific primers and probes. As a positive control, synthetic template at a concentration of 109 cp/mL was spiked into salmonella negative equine feces. Negative sample showed no amplification. Positive pathogen specific detection can be seen for both clinical and spiked samples.

References

  1. Pusterla, N., Byrne, B., Hodzic, E., Mapes, S., Jang, S., & Magdesian, K. (2010). Use of quantitative real-time PCR for the detection of Salmonella spp. in fecal samples from horses at a vetinary teaching hospital. The Veterinary Journal, 186, 252-255.
  2. Stewart, A. J. (2016). Salmonellosis in Horses. Retrieved from Merck Veterinary Manual.