Journal:

Article Title: Genotyping of Enterotoxigenic Clostridium perfringens Fecal Isolates Associated with Antibiotic-Associated Diarrhea and Food Poisoning in North America

doi: 10.1128/JCM.39.3.883-888.2001

Figure Lengend Snippet: Multiplex PCR analysis of North American human GI disease isolates. Representative results shown are for multiplex PCR using primers designed to amplify genes encoding each “typing” toxin and CPE. Migration of PCR products derived from each toxin gene are indicated on the left. As denoted on the figure by the letter shown in parentheses to their right, control typing strains used include NCTC8533 (type B), CN5383 (type C), PS52 (type D), 853 (type E), ATCC 3624 (cpe lacking, type A), and F4969 (cpe-positive, type A). Representative North American human GI disease isolates shown include 537-5 and 538-1 (food poisoning isolates) and T34058 and W30554 (AAD isolates). Molecular sizes of DNA markers are noted on the right of the figure.

Article Snippet: C. perfringens isolates used as controls in this study included F4969, a type A strain carrying the cpe gene on a plasmid ( 8 ); NCTC10239, a type A strain carrying a chromosomal cpe gene ( 8 ); ATCC 3624, a cpe -negative type A isolate ( 10 ); NCTC8533, a cpe -lacking, type B isolate (provided by Richard Titball); CN5383, a cpe -negative type C isolate ( 10 ); PS52, a cpe -negative type D isolate (provided by Ronald Labbe); and 853, a type E isolate carrying silent cpe sequences ( 1 ).

Techniques: Multiplex Assay, Migration, Derivative Assay

Journal:

Article Title: Genotyping of Enterotoxigenic Clostridium perfringens Fecal Isolates Associated with Antibiotic-Associated Diarrhea and Food Poisoning in North America

doi: 10.1128/JCM.39.3.883-888.2001

Figure Lengend Snippet: RFLP Southern blot analysis of NruI-digested DNA from North American human GI disease isolates. Southern blots were probed with a 639-bp DIG-labeled cpe-specific probe. Control isolates shown include 10239 (NCTC10239; a chromosomal cpe, food poisoning isolate) and F4969 (a plasmid cpe, non-food-borne human GI disease isolate). Representative North American human GI disease isolates shown include food poisoning isolates 537-5 and 538-1 and AAD isolates T34058 and W30554. Molecular sizes of DNA markers are given to the left of the blot.

Article Snippet: C. perfringens isolates used as controls in this study included F4969, a type A strain carrying the cpe gene on a plasmid ( 8 ); NCTC10239, a type A strain carrying a chromosomal cpe gene ( 8 ); ATCC 3624, a cpe -negative type A isolate ( 10 ); NCTC8533, a cpe -lacking, type B isolate (provided by Richard Titball); CN5383, a cpe -negative type C isolate ( 10 ); PS52, a cpe -negative type D isolate (provided by Ronald Labbe); and 853, a type E isolate carrying silent cpe sequences ( 1 ).

Techniques: Southern Blot, Labeling, Plasmid Preparation

Journal:

Article Title: Genotyping of Enterotoxigenic Clostridium perfringens Fecal Isolates Associated with Antibiotic-Associated Diarrhea and Food Poisoning in North America

doi: 10.1128/JCM.39.3.883-888.2001

Figure Lengend Snippet: PFGE Southern blot analysis of selected North American human GI disease isolates. PFGE and Southern hybridization analysis of undigested (UC) and I-CeuI cut (C) DNA from selected isolates. Blots are probed with a cpe-specific probe. Control isolates shown include 10239 (NCTC10239; a chromosomal cpe, food poisoning isolate) and F4969 (a plasmid cpe, non-food-borne human GI disease isolate). Representative North American human GI disease isolates shown include food poisoning isolates 537-5 and 538-1 and AAD isolates T34058 and W30554. The pulsed-field gel was calibrated with Lambda DNA markers, whose migration is shown at the left of the blot.

Article Snippet: C. perfringens isolates used as controls in this study included F4969, a type A strain carrying the cpe gene on a plasmid ( 8 ); NCTC10239, a type A strain carrying a chromosomal cpe gene ( 8 ); ATCC 3624, a cpe -negative type A isolate ( 10 ); NCTC8533, a cpe -lacking, type B isolate (provided by Richard Titball); CN5383, a cpe -negative type C isolate ( 10 ); PS52, a cpe -negative type D isolate (provided by Ronald Labbe); and 853, a type E isolate carrying silent cpe sequences ( 1 ).

Techniques: Southern Blot, Hybridization, Plasmid Preparation, Pulsed-Field Gel, Lambda DNA Preparation, Migration

Journal:

Article Title: Genotyping of Enterotoxigenic Clostridium perfringens Fecal Isolates Associated with Antibiotic-Associated Diarrhea and Food Poisoning in North America

doi: 10.1128/JCM.39.3.883-888.2001

Figure Lengend Snippet: Western Blot analysis of CPE expression by selected North American human GI disease isolates. The expression of CPE by sporulating cultures of control and disease isolates of C. perfringens was evaluated using a CPE-specific Western immunoblot procedure. Isolates were grown in sporulation media as described in the Materials and Methods and then sonicated. An aliquot (40 μl) of each sonicated sporulating culture lysate was then subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by immunoblotting with CPE antibodies. The blot was developed for chemiluminescence detection to identify immunoreactive species. Results for control isolates shown include 10239 (NCTC10239, a chromosomal cpe, food poisoning isolate) and F4969 (a plasmid cpe, non-food-borne human GI disease isolate). Results for representative North American human GI disease isolates shown include food poisoning isolates 537-5 and 538-1 and AAD isolates T34058 and W30554. Molecular mass markers are shown at left; the arrow at right indicates the migration of purified CPE.

Article Snippet: C. perfringens isolates used as controls in this study included F4969, a type A strain carrying the cpe gene on a plasmid ( 8 ); NCTC10239, a type A strain carrying a chromosomal cpe gene ( 8 ); ATCC 3624, a cpe -negative type A isolate ( 10 ); NCTC8533, a cpe -lacking, type B isolate (provided by Richard Titball); CN5383, a cpe -negative type C isolate ( 10 ); PS52, a cpe -negative type D isolate (provided by Ronald Labbe); and 853, a type E isolate carrying silent cpe sequences ( 1 ).

Techniques: Western Blot, Expressing, Sonication, Polyacrylamide Gel Electrophoresis, Plasmid Preparation, Migration, Purification