Peer-reviewed veterinary case report
Single-tube nested PCR assay for the detection of avian botulism in cecal contents of chickens.
- Journal:
- Anaerobe
- Year:
- 2015
- Authors:
- Jang, Il et al.
- Affiliation:
- Avian Disease Division · South Korea
Plain-English summary
This study introduces a new test to quickly identify avian botulism, a serious illness in chickens caused by a specific bacteria called Clostridium botulinum types C and C/D. The test uses a method called single-tube nested PCR, which is faster and avoids the ethical concerns of using mice for testing. Researchers confirmed that the test works well by checking it against various bacteria and cecal (intestinal) samples from chickens that had outbreaks of botulism, and it successfully detected the bacteria in all clinical samples tested. In contrast, samples from 300 healthy chickens showed no signs of the bacteria. Overall, this new test is effective, quick, and more cost-efficient than other methods.
Abstract
This paper describes a novel diagnostic method for the detection of avian botulism caused by Clostridium botulinum type C and C/D, using single-tube nested PCR assay. This assay was developed to overcome the disadvantages of bioassays used in experiments with mice. Three primer pairs including an antisense primer were designed to target the N-terminal of the toxin gene from C. botulinum types C and C/D. The specificity of the PCR assay was confirmed by using 33 bacterial strains and chicken cecal contents from farms that experienced botulism outbreaks. The detection limit for purified DNA was 1.1 fg/μl, and for bacterial spores was 4.3 spores/200 mg of cecal contents. While checking for specificity of the PCR assay, the reactions with the templates form C. botulinum type C and C/D which were tested became positive, but the rest of the reactions turned negative. However, the results for all clinical samples (n = 8) were positive. The PCR assay results for cecal samples obtained from 300 healthy chickens (150 Korean native chickens and 150 broilers) were all negative. This assay is rapid and straightforward and evades ethical issues associated with mouse bioassay. Moreover, it is more economical than real-time PCR.
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Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/26159405/