Polymerase Chain Reaction for Detection of Type A Clostridium botulinum in Foods

Ferreira, Joseph; Baumstark, Barbara R.; Hamdy, M. K.; McCay, Steven G.

doi:10.4315/0362-028x-56.1.18

Cited by 21 publications

(11 citation statements)

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“…The prevalence of C. botulinum was 4% (1 of 25) in sediment, 16.6% (29 of 175) in seawater fish, and 25% (1 of 4) in freshwater fish, whereas the 2 shellfish samples and 8 waste (Table 4). PCR toxinotyping identified bont/A in 7 seawater fish samples, bont/B in 21 samples (20 seawater fish and 1 freshwater fish), and bont/E in 2 samples (1 sediment and 1 seawater fish).…”

Section: Resultsmentioning

confidence: 98%

Detection by PCR-Enzyme-Linked Immunosorbent Assay ofClostridium botulinumin Fish and Environmental Samples from a Coastal Area in Northern France

Fach

Perelle

Dilasser

et al. 2002

Appl Environ Microbiol

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The prevalence of Clostridium botulinum types A, B, E, and F was determined in 214 fresh fish and environmental samples collected in Northern France. A newly developed PCR-enzyme-linked immunosorbent assay (ELISA) used in this survey detected more than 80% of samples inoculated with fewer than 10 C. botulinum spores per 25 g and 100% of samples inoculated with more than 30 C. botulinum spores per 25 g. The percent agreement between PCR-ELISA and mouse bioassay was 88.9%, and PCR-ELISA detected more positive samples than the mouse bioassay did. The prevalence of C. botulinum in seawater fish and sediment was 16.6 and 4%, respectively, corresponding to 3.5 to 7 and 1 to 2 C. botulinum most-probable-number counts, respectively, and is in the low range of C. botulinum contamination reported elsewhere. The toxin type identification of the 31 naturally contaminated samples was 71% type B, 22.5% type A, and 9.6% type E. Type F was not detected. The high prevalence of C. botulinum type B in fish samples is relatively unusual compared with the high prevalence of C. botulinum type E reported in many worldwide and northern European surveys. However, fish processing and fish preparation in France have not been identified as a significant hazard for human type B botulism.

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Section: Resultsmentioning

confidence: 98%

Detection by PCR-Enzyme-Linked Immunosorbent Assay ofClostridium botulinumin Fish and Environmental Samples from a Coastal Area in Northern France

Fach

Perelle

Dilasser

et al. 2002

Appl Environ Microbiol

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“…However, mouse bioassay and colony isolation procedures take several days to complete and it is recommended not to use the animals for diagnosis bacause of ethical problems. Recently, the DNA sequences of botulinum type A to G toxin genes have been determined (1-4, 9-11, 13, 14, 16-18), and the polymerase chain reaction (PCR) detecting botulinum type A to G genes has been established (5)(6)(7)15). However, the lengths and the sites of amplified gene fragments were different depending on the studies, and no one used the restriction enzyme digestion profiles of the amplified products for confirming the type of toxin genes.…”

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confidence: 99%

Simple Method for Detection of Clostridium botulinum Type A to F Neurotoxin Genes by Ploymerase Chain Reaction

Takeshi

Fujinaga

Inoue

et al. 1996

Microbiology and Immunology

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A polymerase chain reaction (PCR)-based method was established to detect each type of neurotoxin genes of Clostridium botulinum types A to F by employing the oligonucleotide primer sets corresponding to special regions of the light chains of the neurotoxins. In this procedure, the PCR products were easily confirmed by restriction enzyme digestion profiles, and as little as 2.5 pg of template DNAs from toxigenic strains could be detected. The specific PCR products were obtained from toxigenic C. botulinum types A to F, a type E toxin-producing C. butyricum strain, and a type F toxin-producing C. baratii strain, but no PCR product was detected in nontoxigenic strains of C. botulinum and other clostridial species. The neurotoxin genes were also detected in food products of a seasoned dry salmon and a fermented fish (Izushi) which had caused type E outbreaks of botulism. Therefore, it is concluded that this PCR-based detection method can be used for the rapid diagnosis of botulism.

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“…For the detection of C. botulinum, several detection methods have been reported. The usefulness of PCR was confirmed in food-borne botulism (9,10,32), providing high sensitivity and specificity (17). Several immunoassay systems, another detection method of C. botulinum, were also developed.…”

Section: Resultsmentioning

confidence: 99%

Application of Real‐Time PCR for Quantitative Detection of Clostridium botulinum Type A Toxin Gene in Food

Yoon

Chung

Kang

et al. 2005

Microbiology and Immunology

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The TaqMan real‐time PCR method for the quantitative detection of C. botulinum type A was developed based on sequence‐specific hybridization probes. The validity of this assay was verified by using 10 genera of 20 strains, including reference strains of C. botulinum types A, B, C, D, E and F. The detection limit of this assay was evaluated on C. botulinum type A, using a 10‐fold dilution series of DNA and spores. The DNA and spores were detected up to level of 0.1 ng/ml and 102 spores/ml, respectively. Spore spiked food sample preparation prior to the real‐time PCR was performed by two methods, heat treatment and GuSCN. The detection limits after heat treatment showed 102 spores/ml for spiked sausage slurry, and 103 spores/ml for spiked canned corn slurry, while detection limits after GuSCN precipitation showed 102 spores/ml in both sausage and canned corn. Therefore the real‐time PCR assay after GuSCN precipitation is useful for the quantification of C. botulinum type A because it showed identical CT values in both pure spore solutions and food slurries. We suggest that quantitative analysis of C. botulinum type A by TaqMan real‐time PCR can be a rapid and accurate assessment method for botulinal risk in food samples.

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Polymerase Chain Reaction for Detection of Type A Clostridium botulinum in Foods

Cited by 21 publications

References 0 publications

Detection by PCR-Enzyme-Linked Immunosorbent Assay ofClostridium botulinumin Fish and Environmental Samples from a Coastal Area in Northern France

Detection by PCR-Enzyme-Linked Immunosorbent Assay ofClostridium botulinumin Fish and Environmental Samples from a Coastal Area in Northern France

Simple Method for Detection of Clostridium botulinum Type A to F Neurotoxin Genes by Ploymerase Chain Reaction

Application of Real‐Time PCR for Quantitative Detection of Clostridium botulinum Type A Toxin Gene in Food

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