Detection of Clostridium botulinum type F using the polymerase chain reaction

1994 ◽  
Vol 8 (5) ◽  
pp. 365-373 ◽  
Author(s):  
Joseph L. Ferreira ◽  
Mostafa K. Hamdy ◽  
Steven G. McCay ◽  
Mark Hemphill ◽  
Nameer Kirma ◽  
...  
1996 ◽  
Vol 79 (4) ◽  
pp. 861-865 ◽  
Author(s):  
Carl J Sciacchttano ◽  
Irvin N Hlrshfield

Abstract The polymerase chain reaction (PCR), a rapid, sensitive technique for amplifying target DNA sequences of pathogenic microorgansims, was used to amplify Clostridium botulinum type E neurotoxin gene fragments in smoked fish. Other botulinal neurotoxin-producing strains, nontoxigenic strains, and food-related microorganisms did not yield nonspecificamplification products with this PCR assay. PCR products were analyzed by capillary electrophoresis (CE) using a low-viscosity entangled polymer system. Resolution, sensitivity, and DNA sizing accuracy were improved, and analytical times were markedly shortened. The PCR/CE assay detected the C. botulinum type E neurotoxin gene in as few as 10 cells. The technique to other foods may also be a valuable tool for detecting foodborne pathogens.


2007 ◽  
Vol 54 (8) ◽  
pp. 320-327 ◽  
Author(s):  
V. Prévot ◽  
F. Tweepenninckx ◽  
E. Van Nerom ◽  
A. Linden ◽  
J. Content ◽  
...  

2001 ◽  
Vol 64 (2) ◽  
pp. 201-207 ◽  
Author(s):  
AGNÈS BRACONNIER ◽  
VÉRONIQUE BROUSSOLLE ◽  
SYLVIE PERELLE ◽  
PATRICK FACH ◽  
CHRISTOPHE NGUYEN-THE ◽  
...  

A molecular method was used for the detection of Clostridium botulinum spores of type A, B, and E in commercial cooked and pasteurized vegetable purées and in the raw materials (vegetables and other ingredients). The method allowed the detection of less than 8 spores/g of product for C. botulinum type A, less than 1 spore/g for proteolytic type B, less than 21 spores/g for nonproteolytic type B, and less than 0.1 spore/g for type E. Thirty-seven samples of raw vegetables and ingredients were tested for the presence of C. botulinum type A, B, and E; 88 and 90 samples of vegetable purées were tested, respectively, for the presence of C. botulinum type A and B and for the presence of C. botulinum type E. All samples were negative, suggesting that the prevalence of C. botulinum in these vegetable purées and the raw ingredients is probably low.


2002 ◽  
Vol 85 (5) ◽  
pp. 1025-1028 ◽  
Author(s):  
Kathy E Craven ◽  
Joseph L Ferreira ◽  
Mark A Harrison ◽  
Paul Edmonds

Abstract Clostridium botulinum organisms generally produce 1 of 4 neurotoxin types (A, B, E, and F) associated with human illness. Neurotoxin type determination is important in identification of the bacterium. A polymerase chain reaction (PCR) method was developed to identify 24 h botulinal cultures as potential types A, B, E, and F neurotoxin producers as well as other clostridial species which also produce neurotoxins. Components of the PCR and amplification conditions were adjusted for optimal amplification of toxin gene target regions to enable simultaneous testing for types A, B, E, and F in separate tubes using a single thermal cycler. Each primer set was specific for its corresponding toxin type. A DNA extraction procedure was also included to remove inhibitory substances that may affect amplification. This procedure is rapid, sensitive, and specific for identification of toxigenic C. botulinum.


Author(s):  
Dwiyitno Dwiyitno

Food infection and intoxication of fish products are common due to consumption of foodexposed by either pathogenic or toxin promoter bacteria. A number of pathogenic bacteria havebeen identified related to fish and seafood such as Vibrio parahaemolyticusand other Vibrios,Escherichia coli, Aeromonas spp., Salmonellaspp., Staphylococcus aureus, Listeriamonocytogenes, Clostridium botulinum, C. perfringens, and Shigellaspp. However, isolationand identification of pathogenic bacteria from fish and seafood are often difficult due to the highnumber of contaminating and indigenous bacteria, while numbers of pathogenic bacteria arerelatively low. In the last 3 decades, several molecular methods have been developed and adoptedas official standard to supplement classical method, such as polymerase chain reaction (PCR)and its applications. Besides its limitation, molecular based identification offers advantages asroutine or screening analytical protocol for pathogenic bacteria in fish and seafood products.


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