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ID 56702
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Sinha, A. National Institute of Cholera and Enteric Diseases (NICED)
SenGupta, S. National Institute of Cholera and Enteric Diseases (NICED)
Guin, S. National Institute of Cholera and Enteric Diseases (NICED)
Dutta, S. National Institute of Cholera and Enteric Diseases (NICED)
Ghosh, S. National Institute of Cholera and Enteric Diseases (NICED)
Mukherjee, P. National Institute of Cholera and Enteric Diseases (NICED)
Mukhopadhyay, A. K. National Institute of Cholera and Enteric Diseases (NICED)
Ramamurthy, T. National Institute of Cholera and Enteric Diseases (NICED)
Takeda, Y. Collaborative Research Centre of Okayama University for Infectious Diseases in India, NICED
Kurakawa, T. Yakult Central Institute for Microbiological Research
Nomoto, K. Yakult Central Institute for Microbiological Research
Nair, G. B. National Institute of Cholera and Enteric Diseases (NICED)
Nandy, R. K. National Institute of Cholera and Enteric Diseases (NICED)
Abstract
Culture-independent identification of diarrhoeal aetiological agents was performed using DNA harvested from diarrhoeal stool specimens with SYBR-Green-based real-time PCR targeting Vibrio cholerae, Vibrio parahaemolyticus, Campylobacter spp., Shigella spp. and three different pathotypes of diarrhoeagenic Escherichia coli. Conventional culture-dependent methods detected bacterial enteropathogens in 68 of 122 diarrhoeal stool specimens. Of 68 specimens, 59 (86.8%) had a single pathogen and the remaining nine (13.2%) had polymicrobial infections with multiple pathogens. Re-analysis of the 68 specimens by culture-independent real-time PCR methods showed that 25 (36.8%) specimens contained single pathogen and 43 (63.2%) specimens contained mixed infections with multiple pathogens. The prevalence of such high levels of polymicrobial infections would not have been detected without using real-time PCR. Culture-dependent analysis assigned 54 of the 122 selected archived specimens as 'no known aetiology'. However, re-analysis of these samples by real-time PCR showed the presence of single or multiple pathogens among 34 (63%) of these specimens. Estimation of relative pathogen load by real-time PCR in the stool specimens indicated that the inability of conventional culture-dependent methods to detect the pathogens was related to lower colony-forming units of the pathogen, as reflected by lower C(t) values. Detection of high levels of polymicrobial infection by real-time PCR indicates that in the settings like Kolkata and its surroundings, where cholera and other enteric diseases are endemic, the concept of one pathogen one disease might need to be re-evaluated.
Keywords
Real-time PCR
Diarrhoea
Polymicrobial infection
Note
This is an Accepted Manuscript of an article published by Elsevier B.V
Published Date
2013-02
Publication Title
Clinical Microbiology and Infection
Volume
volume19
Issue
issue2
Publisher
Elsevier B.V
Start Page
173
End Page
180
ISSN
1198743X
NCID
AA11098239
Content Type
Journal Article
language
英語
OAI-PMH Set
岡山大学
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Related Url
isVersionOf https://doi.org/10.1111/j.1469-0691.2011.03746.x
Project
Collaborative Research of Okayama University for Infectious Diseases in India