Detection of bovine Clostridium perfringens by polymerase chain reaction

Piatti, Rosa Maria; Ikuno, A. A.; Baldassi, Lucia

doi:10.1590/s1678-91992004000200005

Cited by 9 publications

(4 citation statements)

References 17 publications

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“…Presence of cpa gene in all the five toxin types (A-E) of C. perfringens was also reported by various other workers (Baums et al, 2004;Piatti et al, 2004). Based on the detection of toxin gene, all the isolates recovered from milk and milk products were found to be of Similarly majority (92.86%) of the isolates recovered from meat and meat products were also found to be C. perfringens type A.…”

Section: Resultssupporting

confidence: 74%

Characterization of enterotoxin producing Clostridium perfringens isolated from foods of animal origin

Haque

Sharma

Barman

et al. 2017

IJAR

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Screening of 105 samples comprising of milk, meat and their products for isolation of Clostridium perfringens revealed a total of 39 (37.14%) samples to be positive for C. perfringens, yielding an equal number of isolates. Among the total isolates, 11 (20.0%) belonged to milk and milk products, of which 10 were isolated from raw cow's milk. The remaining isolate was from paneer sample. Similarly, 28 (56.0%) C. perfringens isolates were recovered from meat and meat products. Majority (37) of the isolates belonged to toxin type A while two isolates recovered each from raw beef and chicken meats were of toxin type C. Among the 39 isolates of C. perfringens, only 5(12.82%) isolates could reveal the presence of cpe (enterotoxin) gene. The protein profile (SDS-PAGE) of both the crude and partially purified enterotoxins prepared from cpe positive C. perfringens isolates (type A and C) revealed a very little difference. Both the partially purified enterotoxins of type A and C of C. perfringens displayed almost similar type of cytopathic effects (CPE) and death of cells in Vero cell line.

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Section: Resultssupporting

confidence: 74%

Characterization of enterotoxin producing Clostridium perfringens isolated from foods of animal origin

Haque

Sharma

Barman

et al. 2017

IJAR

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“…On the other hand, domestic animals are known to be sources of human food poisoning. In order to reduce or eliminate this risk, strategies must be developed to diagnose and prevent infected animals from entering the food chain (10).…”

Section: Introductionmentioning

confidence: 99%

Clostridium perfringens isolate typing by multiplex PCR

Ahsani¹,

Mohammadabadi²,

Shamsaddini³

2010

J. Venom. Anim. Toxins incl. Trop. Dis

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Abstract:Clostridium perfringens is an important pathogen that provokes numerous different diseases. This bacterium is classified into five different types, each of which capable of causing a different disease. There are various methods for the bacterial identification, many are labor-intensive, time-consuming, expensive and also present low sensitivity and specificity. The aim of this research was to identify the different types of C. perfringens using PCR molecular method. In this study, 130 sheep-dung samples were randomly collected from areas around the city of Kerman, southeastern Iran. After processing and culturing of samples, the produced colonies were morphologically studied, gram stain test was also carried out and the genera of these bacteria were identified through biochemical tests. DNA extracted from isolated bacteria for genotyping was tested by multiplex PCR with specific primers. Based on length of synthesized fragments by PCR, toxin types and bacterial strains were detected. C. perfringens isolated types were divided as follows: 17.39% type A, 21.74% type B, 34.78% type C and 26.09% type D. It should be emphasized that, up to the present moment, C. perfringens type A has not been reported in Iran.Key words: Clostridium perfringens, multiplex PCR, biochemical tests, genetic typing. According to Timoney et al. (13), the antitoxin ε was measured in sheep blood sera by ELISA, which proven to be a specific, quick and economical method that may replace the serum neutralization test. ELISA utilizes polyclonal antibodies to identify C. perfringens toxins (14). However, its disadvantage is that the interaction reaction among the produced antibodies works against the toxins, which may difficult the identification of toxin types. Moreover, ELISA falls short of identifying β 2 toxins and in order to identify toxins from spore-forming bacteria, it must be activated by means of special culture methods (15). Original PaPerBiochemical tests are also incapable of distinguishing different types of C. perfringens (16). PCR is the most modern practical technology in diagnosing infectious diseases and compared with classical techniques, it has been shown to be more rapid, with results obtained in a few hours, and also more reliable (12,14). PCR allows a faster bacterial identification directly from clinical samples (17). It should be noted that up to this moment, no research on this subject has been carried out in Iran. MATERIALS AND METHODSIn the present study, sheep dung samples were obtained from 130 Kermani sheep, from nine locations in southeastern Iran. The sheep were from both genders and their ages ranged from 1 to 2.5 years. The samples were collected aseptically in sterile plastic bags and transferred to the laboratory within 1 to 2 hours, after which they were accordingly processed. Samples were diluted in PBS (1:10), the bath temperature was maintained at 80°C for ten minutes in order to eliminate the non-sporeforming bacteria, subsequently they were cultivated on 5% sheep blood agar and anaerobically i...

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“…Organism typing is achieved by culture filtrates with type-specific antisera; these are hard to find, very expensive, time-consuming, and ethically undesirable for tests in experimental animals. The usage of PCR can circumvent these problems and may be used to separate C. perfringens into its five toxin types [21].…”

Section: Molecular Identification Of C Perfringens By Pcr Using Specific Primersmentioning

confidence: 99%

Molecular detection and antimicrobial resistance of Clostridium perfringens isolated from diabetic patients and bullet wounds

2019

jabb

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Clostridium perfringens is a major cause of gas gangrene. The morbidity of C. perfringens is connected with producing toxins. This cross-sectional study was designed to isolate, genetically diagnose, and study the antibiotic susceptibility patterns of C. perfringens isolated from clinical samples. Different wound swabs (from diabetic patients, cellulitis, and bullet wounds) were taken from 140 patients. For isolation of anaerobic bacteria, samples (in thioglycolate broth) were immediately incubated anaerobically then identified according to the cultural properties and biochemical tests. DNA was extracted from all specimens. Polymerase chain reaction was applied for detection of 16SrRNA and internal transcribed spacer (ITS) genes of C. perfringens. The susceptibility of bacterial isolates to different antibiotics was determined using Vitek 2 system and disk diffusion test. Out of 140 clinical samples collected during this study, 3 (2.14%) C. perfringens isolates were recovered of which 2 isolates (1.43%) obtained from diabetic patients and one (0.71%) from bullet wounds. Results also showed that only 7 isolates (5%) were detected by a molecular method using specific primers 16S rRNA and ITS genes of C. perfringens. Results of antibiotic susceptibility testing showed that all isolates were highly susceptible to penicillins and β-lactamase inhibitors, metronidazole, and aminoglycosides. On the other hand, all isolates were highly resistant to tetracycline, levofloxacin, and erythromycin. The susceptibility patterns of C. perfringens isolates showed that all isolates were multidrug resistance. Using the amplification of ITS gene increases specificity and sensitivity (by reducing non-specific annealing and primer dimer formation) which increases the probability of detection of suspected C. perfringens isolates.

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Detection of bovine Clostridium perfringens by polymerase chain reaction

Cited by 9 publications

References 17 publications

Characterization of enterotoxin producing Clostridium perfringens isolated from foods of animal origin

Characterization of enterotoxin producing Clostridium perfringens isolated from foods of animal origin

Clostridium perfringens isolate typing by multiplex PCR

Molecular detection and antimicrobial resistance of Clostridium perfringens isolated from diabetic patients and bullet wounds

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