Toxinotyping of Clostridium perfringens isolates by ELISA and PCR from lambs suspected of enterotoxemia
Hasan Hüseyin Hadimli, Osman ERGANİŞ, Zafer Sayın, Zeki Aras
Abstract
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Hasan Hüseyin Hadimli, Osman ERGANİŞ, Zafer Sayın, Zeki Aras
Abstract
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Clostridium perfringens is a gram-positive, anaerobic bacterium that causes a wide range of diseases in humans and animals. The purposes of this study were to determine, using the enzyme-linked immunosorbent assay (ELISA), the clostridial toxins in intestinal samples of lambs with suspected enterotoxemia; to conduct molecular typing of C. perfringens isolates; and to investigate the presence of the genes of 4 major toxins (\alpha, \beta, \epsilon, and \iota) in the isolates by polymerase chain reaction (PCR). According to the ELISA results, clostridial toxins were determined in 32 intestinal samples. Of the samples, 13 (40.62%) were C. perfringens type A, 1 (3.125%) was C. perfringens type B, 9 (28.125%) were C. perfringens type C, and 9 (28.125%) were C. perfringens type D. C. perfringens was isolated from 13 (8.66%) out of 150 intestinal samples. A total of 13 C. perfringens isolates from lambs were genotyped by PCR. Of the isolates, 10 (76.92%) were type A, 2 (%15.38) were type D, and 1 (7.69%) was type C. Type B and type E were not identified. The results showed that C. perfringens type A was the most common type by both ELISA and PCR in lambs with enterotoxemia.
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Clostridium perfringens is a gram-positive, anaerobic bacterium that causes a wide range of diseases in humans and animals. The purposes of this study were to determine, using the enzyme-linked immunosorbent assay (ELISA), the clostridial toxins in intestinal samples of lambs with suspected enterotoxemia; to conduct molecular typing of C. perfringens isolates; and to investigate the presence of the genes of 4 major toxins (\alpha, \beta, \epsilon, and \iota) in the isolates by polymerase chain reaction (PCR). According to the ELISA results, clostridial toxins were determined in 32 intestinal samples. Of the samples, 13 (40.62%) were C. perfringens type A, 1 (3.125%) was C. perfringens type B, 9 (28.125%) were C. perfringens type C, and 9 (28.125%) were C. perfringens type D. C. perfringens was isolated from 13 (8.66%) out of 150 intestinal samples. A total of 13 C. perfringens isolates from lambs were genotyped by PCR. Of the isolates, 10 (76.92%) were type A, 2 (%15.38) were type D, and 1 (7.69%) was type C. Type B and type E were not identified. The results showed that C. perfringens type A was the most common type by both ELISA and PCR in lambs with enterotoxemia.
Key concepts: Clostridium perfringens, Microbiology, Biology, Polymerase chain reaction, Typing, Virology, Bacteria, Gene