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A filter immunobinding technique for the rapid detection and simultaneous identification of avian and bovine
T Takahata1, M Kato, H Nagatomo
1Animal Health Products and Chemicals Research Center, Daiichi Pharmaceutical Co., Ltd., Tokyo, Japan.
Abstract:
A filter immunobinding (FIB) method was developed for the detection and identification of mycoplasmas. Type strains of a total of 18 avian and bovine mycoplasma species propagated in broth media were diluted and immobilized on a nitrocellulose membrane as antigens for investigating the specificity with rabbit hyperimmune sera. Non-specific FIB reactions were easily eliminated by the procedure of absorbing rabbit hyperimmune sera in the broth. Absorbed rabbit hyperimmune sera exhibited clear species-specificity with mycoplasma antigens by the FIB. These specific reactions always agreed with the results of identification by tests of biochemical properties and growth inhibition for the isolates of M. bovirhinis, M. bovis, M. columbinum, M. columborale, M. gallisepticum and M. synoviae. Some bovine mycoplasma species, which were impossible to identify by growth inhibition test, because of their strong production of film and spots on the agar, specifically reacted with absorbed rabbit hyperimmune sera against M. bovis in the FIB. The detection limit of mycoplasmas by this method was about 10(4)-10(5) colony-forming units/ml, which is lower than that of colony determination on agar. The FIB seems to be a useful technique for rapid detection and simultaneous identification of mycoplasmas.
Insights
A new filter immunobinding (FIB) method rapidly detects and identifies avian and bovine mycoplasmas. This technique offers species-specific results, improving upon traditional methods for mycoplasma detection.
Area of Science:
- Veterinary Microbiology
- Immunodiagnostics
- Bacteriology
Background:
- Mycoplasma species cause significant diseases in avian and bovine populations.
- Accurate and rapid detection of mycoplasmas is crucial for disease control and management.
- Existing diagnostic methods like growth inhibition can be challenging for certain Mycoplasma species.
Purpose of the Study:
- To develop and validate a novel filter immunobinding (FIB) method for mycoplasma detection.
- To assess the specificity and sensitivity of the FIB method for identifying avian and bovine mycoplasma species.
- To compare the FIB method with conventional diagnostic techniques.
Main Methods:
- Development of a filter immunobinding (FIB) assay using nitrocellulose membranes.
- Immobilization of mycoplasma type strains as antigens.
- Use of absorbed rabbit hyperimmune sera for specific antibody detection.
- Validation against biochemical tests and growth inhibition assays.
Main Results:
- The FIB method demonstrated clear species-specificity for 18 avian and bovine mycoplasma species.
- Non-specific reactions were eliminated by absorbing hyperimmune sera.
- FIB results correlated well with biochemical and growth inhibition tests for several species.
- The FIB method successfully identified bovine mycoplasma species that were difficult to diagnose using growth inhibition.
- The detection limit was approximately 10(4)-10(5) colony-forming units/ml, surpassing agar-based colony counts.
Conclusions:
- The filter immunobinding (FIB) assay is a reliable and specific method for detecting and identifying mycoplasmas.
- FIB offers advantages over traditional methods, particularly for challenging Mycoplasma species.
- This technique provides a useful tool for the rapid and simultaneous diagnosis of mycoplasma infections.