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Sensitive quantification of Clostridium perfringens in human feces by quantitative real-time PCR targeting alpha-toxin and enterotoxin genes

Overview of attention for article published in BMC Microbiology, October 2015
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Title
Sensitive quantification of Clostridium perfringens in human feces by quantitative real-time PCR targeting alpha-toxin and enterotoxin genes
Published in
BMC Microbiology, October 2015
DOI 10.1186/s12866-015-0561-y
Pubmed ID
Authors

Ravinder Nagpal, Kiyohito Ogata, Hirokazu Tsuji, Kazunori Matsuda, Takuya Takahashi, Koji Nomoto, Yoshio Suzuki, Kazunari Kawashima, Satoru Nagata, Yuichiro Yamashiro

Abstract

Clostridium perfringens is a widespread pathogen, but the precise quantification of this subdominant gut microbe remains difficult due to its low fecal count (particularly in asymptomatic subjects) and also due to the presence of abundant polymerase-inhibitory substances in human feces. Also, information on the intestinal carriage of toxigenic C. perfringens strains in healthy subjects is sparse. Therefore, we developed a sensitive quantitative real-time PCR assays for quantification of C. perfringens in human feces by targeting its α-toxin and enterotoxin genes. To validate the assays, we finally observed the occurrence of α-toxigenic and enterotoxigenic C. perfringens in the fecal microbiota of healthy Japanese infants and young adults. The plc-specific qPCR assay was newly validated, while primers for 16S rRNA and cpe genes were retrieved from literature. The assays were validated for specificity and sensitivity in pre-inoculated fecal samples, and were finally applied to quantify C. perfringens in stool samples from apparently healthy infants (n 124) and young adults (n 221). The qPCR assays were highly specific and sensitive, with a minimum detection limit of 10(3) bacterial cells/g feces. Alpha-toxigenic C. perfringens was detected in 36 % infants and 33 % adults, with counts ranging widely (10(3)-10(7) bacterial cells/g). Intriguingly, the mean count of α-toxigenic C. perfringens was significantly higher in infants (6.0 ± 1.5 log10 bacterial cells/g), as compared to that in adults (4.8 ± 1.2). Moreover, the prevalence of enterotoxigenic C. perfringens was also found to be significantly higher in infants, as compared to that in adults. The mean enterotoxigenic C. perfringens count was 5.9 ± 1.9 and 4.8 ± 0.8 log10 bacterial cells/g in infants and adults, respectively. These 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.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 82 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Japan 1 1%
Unknown 81 99%

Demographic breakdown

Readers by professional status Count As %
Researcher 19 23%
Student > Ph. D. Student 10 12%
Student > Master 9 11%
Student > Bachelor 7 9%
Student > Doctoral Student 5 6%
Other 15 18%
Unknown 17 21%
Readers by discipline Count As %
Agricultural and Biological Sciences 20 24%
Immunology and Microbiology 9 11%
Medicine and Dentistry 9 11%
Biochemistry, Genetics and Molecular Biology 7 9%
Veterinary Science and Veterinary Medicine 5 6%
Other 8 10%
Unknown 24 29%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 21 October 2015.
All research outputs
#20,294,248
of 22,830,751 outputs
Outputs from BMC Microbiology
#2,689
of 3,191 outputs
Outputs of similar age
#237,871
of 283,771 outputs
Outputs of similar age from BMC Microbiology
#56
of 75 outputs
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So far Altmetric has tracked 3,191 research outputs from this source. They receive a mean Attention Score of 4.1. This one is in the 1st percentile – i.e., 1% of its peers scored the same or lower than it.
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