Sensitive quantification of Clostridium perfringens

Clostridium perfringens is one of the most widely dispersed opportunistic pathogens and is well known to produce a number of toxins to cause several forms of histotoxic and enteric diseases in humans and animals [2]. Based on the production of four major toxins i.e., alpha, beta, epsilon and iota, it is categorized into five toxin-types viz. A, B, C, D and E. While it is ambiguous why C. perfringens produces so many diverse toxins, it is well known that it uses chromosomally-encoded α-toxin (which has phospholipase C (plc) and sphingomyelinase activities with hemolytic, necrotic and lethal abilities) as a chief virulent factor and key mediator for most of C. perfringens-associated diseases.”

Standard curves representing the quantitative detection of reference strains of C. perfringens by Amp-qPCR assay. C. perfringens ATCC 13124T, ATCC 9856, ATCC 3624, ATCC 3626, ATCC 12917, ATCC 14809, ATCC 27324, and CS 052–1 were cultivated separately in Glu-mGAM. DNA fractions were extracted from the culture samples in the early stationary phase (24 h), and bacterial counts were determined microscopically with DAPI staining. 10-fold serial dilutions of DNA corresponding to the bacterial counts ranging from 100 to 105 bacterial cells were assessed by 16S rRNA gene-specific aplc-specific b, and cpe-specific c Amp-qPCR assays. The Cq values obtained were plotted against the log10number of bacterial cells subjected to each reaction. Data are expressed as means and standard deviations of the results from 7 strains (ATCC 13124T, ATCC 9856, ATCC 3624, ATCC 3626, ATCC 12917, ATCC 14809, and ATCC 27324) in the 16S rRNA gene-specific and plc-specific primer sets, and 3 strains (ATCC 12917, ATCC 14809, and CS 052–1) in the cpe-specific primer set

‘This study data indicate that some healthy infants and young adults carry α-toxigenic and enterotoxigenic C. perfringens at significant levels, and may be predisposed to related diseases. Thus, high fecal carriage of toxigenic C. perfringens in healthy children warrants further investigation on its potential sources and clinical significance in these subjects. In summary, we present a novel qPCR assay for sensitive and accurate quantification of α-toxigenic and enterotoxigenic C. perfringens in human feces, which should facilitate prospective studies of the gut microbiota.”

MacFie J. Current status of bacterial translocation as a cause of surgical sepsis. Br Med Bull. 2004 Dec 13;71:1-11. Free Full Text

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