A 16S rDNA-based PCR method for rapid and specific detection of Clostridium perfringens in food

Wang, Rongfu; Cao, Wei‐Wen; Franklin, Wirt; Campbell, Warren L.; Cerniglia, Carl E.

doi:10.1006/mcpr.1994.1018

Cited by 84 publications

(44 citation statements)

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“…We believe that the gyrB region will have high reliability for identifying pathogenic bacteria. Although the 16S rRNA sequence method is a highly accurate and rapid method for identifying most bacteria to the genus level (33), the gyrB sequence method might be more useful for identifying bacteria to the species level. In practical terms, this means that whereas it is easy to amplify the 16S rRNA sequence because it is highly conserved across bacteria, it is harder to amplify the gyrB region because of its variability.…”

Section: Discussionmentioning

confidence: 99%

Phylogenetic Analysis of Salmonella , Shigella , and Escherichia coli Strains on the Basis of the gyrB Gene Sequence

Fukushima¹,

Kakinuma²,

Kawaguchi³

2002

J Clin Microbiol

166

101

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Phylogenetic analysis of about 200 strains of Salmonella, Shigella, and Escherichia coli was carried out using the nucleotide sequence of the gene for DNA gyrase B (gyrB), which was determined by directly sequencing PCR fragments. The results establish a new phylogenetic tree for the classification of Salmonella, Shigella, and Escherichia coli in which Salmonella forms a cluster separate from but closely related to Shigella and E. coli. In comparison with 16S rRNA analysis, the gyrB sequences indicated a greater evolutionary divergence for the bacteria. Thus, in screening for the presence of bacteria, the gyrB gene might be a useful tool for differentiating between closely related species of bacteria such as Shigella spp. and E. coli. At present, 16S rRNA sequence analysis is an accurate and rapid method for identifying most unknown bacteria to the genus level because the highly conserved 16S rRNA region is easy to amplify; however, analysis of the more variable gyrB sequence region can identify unknown bacteria to the species level. In summary, we have shown that gyrB sequence analysis is a useful alternative to 16S rRNA analysis for constructing the phylogenetic relationships of bacteria, in particular for the classification of closely related bacterial species.

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Section: Discussionmentioning

confidence: 99%

Phylogenetic Analysis of Salmonella , Shigella , and Escherichia coli Strains on the Basis of the gyrB Gene Sequence

Fukushima¹,

Kakinuma²,

Kawaguchi³

2002

J Clin Microbiol

166

101

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“…We successfully detected and genotyped C. perfringens in tissue sections from two autopsy cases.Clostridium perfringens causes several forms of enteric diseases, including food poisoning and fatal enterotoxemia (15,17). Based on the presence of four major lethal toxins (alpha-, beta-, epsilon-, and iota-toxins), C. perfringens is classified into five toxigenic types (A through E), and each type can cause different diseases.…”

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confidence: 99%

“…Y12669) (15) and to the alpha-toxin gene or cpa (GenBank accession no. X13608) (8), beta-toxin gene or cpb (GenBank accession no.…”

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confidence: 99%

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Detection and Toxin Typing ofClostridium perfringensin Formalin-Fixed, Paraffin-Embedded Tissue Samples by PCR

Zhang

Xie

et al. 2009

J Clin Microbiol

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Since current microbiology methods are not suitable to detect Clostridium perfringens in formalin-fixed, paraffin-embedded tissue samples, we developed a PCR assay to detect toxin-encoding genes and the 16S rRNA gene of C. perfringens. We successfully detected and genotyped C. perfringens in tissue sections from two autopsy cases.Clostridium perfringens causes several forms of enteric diseases, including food poisoning and fatal enterotoxemia (15,17). Based on the presence of four major lethal toxins (alpha-, beta-, epsilon-, and iota-toxins), C. perfringens is classified into five toxigenic types (A through E), and each type can cause different diseases. The most commonly encountered type A (alpha-toxin) strain causes gas gangrene (myonecrosis), diarrhea, and food-borne illness in humans (4). Type B (alpha-, beta-, and epsilon-toxins) and type D (alpha-and epsilontoxins) strains are the causative agents of fatal enterotoxemia in domestic animals and occasionally humans (16).

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“…In addition, these isolates were examined using the PCR method with 2 kinds of species-specific primers for the 16S rRNA genes of C. perfringens (13,14).…”

Section: Samples (I) Outbreakmentioning

confidence: 99%

Four Foodborne Disease Outbreaks Caused by a New Type of Enterotoxin-Producing Clostridium perfringens

et al. 2015

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The epidemiological and bacteriological investigations on four foodborne outbreaks caused by a new type of enterotoxinproducing Clostridium perfringens are described. C. perfringens isolated from patients of these outbreaks did not produce any known enterotoxin and did not carry the C. perfringens enterotoxin gene. However, the culture filtrates of these isolates induced the accumulation of fluid in rabbit ileal loop tests. The molecular weight of the new enterotoxin may be between 50,000 and 100,000, although the known C. perfringens enterotoxin is ca. 35,000. This new enterotoxin was heat labile, and its biological activities were inactivated by heating for 5 min at 60°C. The new enterotoxin was sensitive to pH values higher than 11.0 and protease treatment but was resistant to trypsin treatment. These results suggest that the new enterotoxin may be a protein. Although C. perfringens enterotoxin induced morphological changes in Vero cells, the changes induced by the new enterotoxin differed from those by the known C. perfringens enterotoxin. The new enterotoxin also induced morphological changes in L929 cells, whereas the known C. perfringens enterotoxin did not, because L929 cells lacked an appropriate enterotoxin receptor. Although C. perfringens enterotoxin is recognized as the only diarrheagenic toxin responsible for C. perfringens foodborne outbreaks, the results of the present study indicate that C. perfringens isolated from these four outbreaks produced a new type of enterotoxin.

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A 16S rDNA-based PCR method for rapid and specific detection of Clostridium perfringens in food

Cited by 84 publications

References 0 publications

Phylogenetic Analysis of Salmonella , Shigella , and Escherichia coli Strains on the Basis of the gyrB Gene Sequence

Phylogenetic Analysis of Salmonella , Shigella , and Escherichia coli Strains on the Basis of the gyrB Gene Sequence

Detection and Toxin Typing ofClostridium perfringensin Formalin-Fixed, Paraffin-Embedded Tissue Samples by PCR

Four Foodborne Disease Outbreaks Caused by a New Type of Enterotoxin-Producing Clostridium perfringens

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