DETECTION OF ALPHA TOXIN PRODUCED BY LOCALLY ISOLATED   CLOSTRIDIUM PERFRINGENS

	DETECTION OF ALPHA TOXIN PRODUCED BY LOCALLY ISOLATED CLOSTRIDIUM PERFRINGENS
Iraqi Journal of Biotechnology
Article 1, Volume 9, Issue 3, 2010, Pages 447-460
Abstract
Locally isolated and diagnosed C.perfringes (26 isolates) had been tested for alpha toxin producing using Trypticase Yeast extract Broth (TYB) .Crude protein concentration of supernatant of these isolates was estimated and the results showed were variability in concentration ranging between 1.162 µg /ml for isolate number (3) wich was high protein produer to 0.01 µg /ml for isolate number (23) with low protein production. The ability of alpha toxin production was first tested by two routin methods, diffusion in egg yolk media and measurement of fluoridation in liquefied egg yolk. The result showed that (24)(92.31%) isolates gave positive test. Mice lethality method was depended to show whether these isolates were toxic. It was appeared that 25 isolates (96.15%). Enzyme linked immune sorbant assay ELISA test was used to detect bacterial ability for alpha toxin production. Results showed that all isolates were positive for toxin production, and all isolates were toxin producer as compared with the routine methods. The product of amplification of cpa gene coded for alpha toxin was then used in detecting and diagnosis of C. perfingens using polymerase chain reaction (PCR). The results showed that all isolates gave positive results in both of bacterial and toxin detection specially when DNA template prepared by boiling was used instead of usual extraction method .
Keywords
Clostridium perfringens; Alpha toxin; ELISA; PCR

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