Clostridium perfringens type A enterotoxin: a rapid method for preparation of a specific antiserum using an enterotoxin purified by the polyacrylamide gel electrophoresis technique

Popoff, M

doi:10.1016/0378-1097(84)90168-x

Cited by 4 publications

(4 citation statements)

References 12 publications

(17 reference statements)

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“…The duplex PCR procedure data indicated that C. perfringens 8-6 was enterotoxigenic ( Table 1). These data are consistent with the fact that this strain produced CPE, as determined by Vero cell cytoxicity, mouse lethality, and immunoprecipitation (22). The other C. perfringens strains were not tested for CPE production, since they did not sporulate under the experimental conditions used.…”

Section: Perfringens Duplex Pcr Specificity and Characterization Osupporting

confidence: 80%

Detection of enterotoxigenic Clostridium perfringens in food and fecal samples with a duplex PCR and the slide latex agglutination test

Fach

Popoff

1997

Appl Environ Microbiol

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A duplex PCR procedure was evaluated for the detection of Clostridium perfringens in food and biological samples and for the identification of enterotoxigenic strains. This method uses two sets of primers which amplify in the same reaction two different DNA fragments simultaneously: the 283-bp C. perfringens phospholipase C gene fragment and the 426-bp enterotoxin gene fragment. Internal primers within the two primer sets confirmed the specificity of the method by DNA-DNA hybridization with the PCR products. No cross-reaction was observed with other Clostridium species or with other bacteria routinely found in food. The detection level was approximately 10 5 C. perfringens cells per g of stool or food sample. When overnight enrichment culture was used, 10 C. perfringens cells per g was detected in 57 artificially contaminated food samples. The duplex PCR is a rapid, sensitive, and reliable method for the detection and identification of enterotoxigenic C. perfringens strains in food samples. A slide latex agglutination test was also evaluated as a rapid, simple technique for the detection of C. perfringens enterotoxin in stool samples.

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Section: Perfringens Duplex Pcr Specificity and Characterization Osupporting

confidence: 80%

Detection of enterotoxigenic Clostridium perfringens in food and fecal samples with a duplex PCR and the slide latex agglutination test

Fach

Popoff

1997

Appl Environ Microbiol

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“…Virulent Clostridium sordellii strains are responsible for lethal infections in several animal species, including enteritis and enterotoxaemia in sheep, lamb and cattle (Richards and Hunt, ; Al‐Mashat and Taylor, ; Popoff, ; Lewis and Naylor, ; Clark, ), equine myopathy (Unger‐Torroledo et al ., ), and myonecrosis and gangrene (Cunniffe, ; Pfeifer et al ., ; Rupnik et al ., ) in humans often accompanied by a toxic shock syndrome. In the last decade, several fatal toxic shock syndromes in women were reported that occurred postpartum and after spontaneous or abortion induced with RU‐486 (Bitti et al ., ; Rorbye et al ., ; Sinave et al ., ; Fischer et al ., ; Couzin, ; Winikoff, ; Cohen et al ., ; Soper, ; Agrawal and Garg, ).…”

Section: Introductionmentioning

confidence: 99%

Haemorrhagic toxin and lethal toxin fromClostridium sordelliistrain vpi9048: molecular characterization and comparative analysis of substrate specificity of the large clostridial glucosylating toxins

et al. 2014

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SummaryLarge clostridial glucosylating toxins (LCGTs) are produced by toxigenic strains of Clostridium difficile, Clostridium perfringens, Clostridium novyi and Clostridium sordellii. While most C. sordellii strains solely produce lethal toxin (TcsL), C. sordellii strain VPI9048 co-produces both hemorrhagic toxin (TcsH) and TcsL. Here, the sequences of TcsH-9048 and TcsL-9048 are provided, showing that both toxins retain conserved LCGT features and that TcsL and TcsH are highly related to Toxin A (TcdA) and Toxin B (TcdB) from C. difficile strain VPI10463. The substrate profile of the toxins was investigated with recombinant LCGT transferase domains (rN) and a wide panel of small GTPases. rN-TcsH-9048 and rN-TcdA-10463 glucosylated preferably Rho-GTPases but also Ras-GTPases to some extent. In this respect, rN-TcsH-9048 and rN-TcdA-10463 differ from the respective full-length TcsH-9048 and TcdA-10463, which exclusively glucosylate Rho-GTPases. rNTcsL-9048 and full length TcsL-9048 glucosylate both Rho-and Ras-GTPases, whereas rN-TcdB-10463 and full length TcdB-10463 exclusively glucosylate Rho-GTPases. Vero cells treated with full length TcsH-9048 or TcdA-10463 also showed glucosylation of Ras, albeit to a lower extent than of Rho-GTPases. Thus, in vitro analysis of substrate spectra using recombinant transferase domains corresponding to the auto-proteolytically cleaved domains, predicts more precisely the in vivo substrates than the full length toxins. Except for TcdB-1470, all LCGTs evoked increased expression of the small GTPase RhoB, which exhibited cytoprotective activity in cells treated with TcsL isoforms, but pro-apoptotic activity in cells treated with TcdA, TcdB, and TcsH. All LCGTs induced a rapid dephosphorylation of pY118-paxillin and of pS144/141-PAK1/2 prior to actin filament depolymerization indicating that disassembly of focal adhesions is an early event leading to the disorganization of the actin cytoskeleton.

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“…The toxin is produced during sporulation (7,16) and it has been purified (1,8,12,(20)(21)(22)(23)(24)27) from sonic extracts of sporulated bacteria and shown to be a simple protein with a molecular weight of ca. 35000 (11,13,14,25).…”

Section: Introductionmentioning

confidence: 99%

Purification by high performance liquid chromatography of Clostridium perfringens type a enterotoxin prepared from high toxin producers selected by a toxin-antitoxin halo

1985

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High enterotoxin-producing substrains of Clostridium perfringens type A were selected reproducibly as colonies having toxin-antitoxin haloes on agar plates of Duncan-Strong medium containing antitoxin serum. Enterotoxin from these substrains was subjected to rapid purification by high performance liquid chromatography (HPLC). For this, the toxin was extracted by sonication from sporulating bacteria grown in Duncan-Strong sporulation medium, fractionated by ammonium sulfate (40% saturation) precipitation and differential solubilization and then purified by HPLC: gel permeation chromatography through a G2000SW column and ion-exchange chromatography on a Mono Q column. Purified toxin preparations had a similar specific activity (4.2 X 10(2) mouse MLD/mg protein) and homogeneity on polyacrylamide gel-electrophoresis to preparations obtained by conventional gel permeation through a Sephadex-G200 column. By further HPLC on a Mono Q column, minor nontoxin proteins were separated from the toxin without loss of the toxicity on a protein basis. The final yield of the purified toxin was about 15% of that in the bacterial extract. The two HPLC procedures each took only one hour.

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Clostridium perfringens type A enterotoxin: a rapid method for preparation of a specific antiserum using an enterotoxin purified by the polyacrylamide gel electrophoresis technique

Cited by 4 publications

References 12 publications

Detection of enterotoxigenic Clostridium perfringens in food and fecal samples with a duplex PCR and the slide latex agglutination test

Detection of enterotoxigenic Clostridium perfringens in food and fecal samples with a duplex PCR and the slide latex agglutination test

Haemorrhagic toxin and lethal toxin fromClostridium sordelliistrain vpi9048: molecular characterization and comparative analysis of substrate specificity of the large clostridial glucosylating toxins

Purification by high performance liquid chromatography of Clostridium perfringens type a enterotoxin prepared from high toxin producers selected by a toxin-antitoxin halo

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