clostridium perfringens netF cytotoxin

Background

C. perfringens is a gram-positive, rod-shaped, anaerobic, spore-forming bacterium that is classified into 5 toxinotypes (A, NetF[Please add this to all C.perf on website, as highlighted. ] , B, C, D, and E) according to the 4 major toxins they produce.1 The toxins are common causes of diarrhea, enterocolitis (inflammation of the small and large intestine), enterotoxemia, and colitis (inflammation of the colon) in horses, goats, pigs, and cattle.2 The NetF toxin is one of a series of recently found toxins produced by the spore forming bacteria. Normally, C. perfringens are nonpathogenic and present in the gastrointestinal tract in low numbers. However, antibiotics use, dietary imbalances, and other stressors may cause overgrowth of C. perfringens, leading to GI diseases.3 The disease may occur sporadically or as an outbreak, and clinically affected neonatal foals have high risk of rapid death. Because nonpathogenic serotypes are common, diagnosis is based on identification of toxigenic C. perfringens in fecal samples. PCR detection for toxin-producing genes is recommended.4 Fluxergy’s rapid PCR test is toxin specific and provides reliable detection of the NetF cytotoxin in a point-of-care format.

Key Benefits

Simple workflow

No sample preparation

Automated data interpretation

Sample to result in 30 minutes

Early detection is treatable with antibiotics

Toxin specific

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 C. perfringens NetF Buffer 1
  • Fluxergy C. perfringens NetF Buffer 2
  • Fluxergy Card

Customer supplied reagents and supplies

  • Clinical sample for testing

Sample collection method

For, 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. Place sample at 4°C if not testing immediately.

Intended use

The Fluxergy C. perfringens NetF assay is a qualitative test for the rapid detection of C. perfringens 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 C. perfringens NetF assay is for Research Use Only (RUO). It is not intended for diagnostic use.
  • Fluxergy’s C. perfringens NetF 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 C. perfringens NetF assay is compatible only for equine fecal swab. The sample type must be specified as each buffer is specially crafted for the sample.
  • Contamination of the sample and kit contents may lead to erroneous results. Use aseptic technique and a clean workspace whenever possible.
  • 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

Clinical sample detection. A clinical equine fecal sample tested positive for C. perfringens NetF, as indicated by amplification and positive agreement with the clinical reference laboratory assay. This result was obtained using the Fluxergy Analyzer Beta and C. perfringens NetF assay.

References

  1. Uzal, F. A., Vidal, J. E., McClane, B. A., & Gurjar, A. A. (2010). Clostridium perfringens toxins involved in mammalian veterinary diseases. The open toxinology journal, 2, 24.
  2. Silva, R. O. S., Oliveira Junior, C. A., Guedes, R. M. C., & Lobato, F. C. F. (2015). Clostridium perfringens: a review of the disease in pigs, horses and broiler chickens. Ciência Rural, 45(6), 1027-1034.
  3. Traub-Dargatz, J. L., & Jones, R. L. (1993). Clostridia-associated enterocolitis in adult horses and foals. Veterinary Clinics of North America: Equine Practice, 9(2), 411-421.
  4. Allison J. Stewart, Clostridia-associated Enterocolitis in Horses. Merck Veterinary Manual.