Detection of Clostridium botulinum Type C Cells in the Gastrointestinal Tracts of Mozambique Tilapia (Oreochromis mossambicus) by Polymerase Chain Reaction

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.

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Microbiology of the food chain. Polymerase chain reaction (PCR) for the detection of food-borne pathogens. Detection of botulinum type A, B, E and F neurotoxin-producing clostridia

10.3403/30261089u ◽

2015 ◽

Keyword(s):

Polymerase Chain Reaction ◽

Food Chain ◽

Type A ◽

Chain Reaction ◽

Food Borne Pathogens ◽

Food Borne ◽

Botulinum Type ◽

Polymerase Chain

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Molecular typing of Clostridium perfringens isolated from swine in slaughterhouses from São Paulo State, Brazil

Ciência Rural ◽

10.1590/s0103-84782012000800020 ◽

2012 ◽

Vol 42 (8) ◽

pp. 1450-1456 ◽

Cited By ~ 1

Author(s):

Thais Sebastiana Porfida Ferreira ◽

Andrea Micke Moreno ◽

Renata Rodrigues de Almeida ◽

Cleise Ribeiro Gomes ◽

Debora Dirani Sena de Gobbi ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Clostridium Perfringens ◽

Food Poisoning ◽

Positive Bacterium ◽

Chain Reaction ◽

High Prevalence ◽

Fecal Isolate ◽

Finishing Pig ◽

Type C ◽

Polymerase Chain

Clostridium perfringens is an anaerobic Gram-positive bacterium known as common pathogen for humans, for domestic and wildlife animals. Although infections caused by C. perfringens type C and A in swine are well studied, just a few reports describe the genetic relationship among strains in the epidemiological chain of swine clostridioses, as well as the presence of the microorganism in the slaughterhouses. The aim of the present study was to isolate C. perfringens from feces and carcasses from swine slaughterhouses, characterize the strains in relation to the presence of enterotoxin, alpha, beta, epsilon, iota and beta-2 toxins genes, using polymerase chain reaction (PCR) and comparing strains by means of Pulsed field gel electrophoresis (PFGE). Clostridium perfringens isolation frequencies in carcasses and finishing pig intestines were of 58.8% in both types of samples. According to the polymerase chain reaction assay, only alfa toxin was detected, being all isolates also negative to enterotoxin and beta2 toxin. Through PFGE technique, the strains were characterized in 35 pulsotypes. In only one pulsotype, the isolate from carcass sample was grouped with fecal isolate of the same animal, suggesting that the risk of cross-contamination was low. Despite the high prevalence of C. perfringens in swine carcasses from the slaughterhouses assessed, the risk of food poisoning to Brazilian pork consumers is low, since all strains were negative to cpe-gene, codifying enterotoxin.

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Screening for Clostridium botulinum Type A, B, and E in Cooked Chilled Foods Containing Vegetables and Raw Material Using Polymerase Chain Reaction and Molecular Probes

Journal of Food Protection ◽

10.4315/0362-028x-64.2.201 ◽

2001 ◽

Vol 64 (2) ◽

pp. 201-207 ◽

Cited By ~ 24

Author(s):

AGNÈS BRACONNIER ◽

VÉRONIQUE BROUSSOLLE ◽

SYLVIE PERELLE ◽

PATRICK FACH ◽

CHRISTOPHE NGUYEN-THE ◽

...

Keyword(s):

Clostridium Botulinum ◽

Raw Materials ◽

Type A ◽

Molecular Probes ◽

Raw Material ◽

Type B ◽

Molecular Method ◽

Chain Reaction ◽

Botulinum Type ◽

Polymerase Chain

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.

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Specific Detection of Clostridium botulinum Types A, B, E, and F Using the Polymerase Chain Reaction

Journal of AOAC International ◽

10.1093/jaoac/85.5.1025 ◽

2002 ◽

Vol 85 (5) ◽

pp. 1025-1028 ◽

Cited By ~ 8

Author(s):

Kathy E Craven ◽

Joseph L Ferreira ◽

Mark A Harrison ◽

Paul Edmonds

Keyword(s):

Polymerase Chain Reaction ◽

Clostridium Botulinum ◽

Extraction Procedure ◽

Toxin Gene ◽

Chain Reaction ◽

Gene Target ◽

Human Illness ◽

Polymerase Chain ◽

Clostridial Species ◽

Thermal Cycler

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.

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