Detection of multiresistant Salmonella typhimurium DT104 using multiplex and fluorogenic PCR

S A Carlson; L F Bolton; C E Briggs; H S Hurd; V K Sharma; P J Fedorka-Cray; B D Jones

doi:10.1006/mcpr.1999.0240

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Detection of multiresistant Salmonella typhimurium DT104 using multiplex and fluorogenic PCR

Journal article

Peer reviewed

Detection of multiresistant Salmonella typhimurium DT104 using multiplex and fluorogenic PCR

S A Carlson, L F Bolton, C E Briggs, H S Hurd, V K Sharma, P J Fedorka-Cray and B D Jones

Molecular and cellular probes, Vol.13(3), pp.213-222

06/1999

DOI: 10.1006/mcpr.1999.0240

PMID: 10369747

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Abstract

Salmonella infections continue to cause gastrointestinal and systemic disease throughout the world. Salmonella typhimurium DT104 further poses a major health concern due to its acquisition of resistance to multiple antibiotics. The rapid detection of multiresistant S. typhimurium DT104 would facilitate strategies aimed at controlling this pathogen. We developed a specific and sensitive polymerase chain reaction (PCR) assay that amplifies a segment of DNA that is conserved in multiresistant S. typhimurium DT104. To provide further specificity for this PCR-based diagnostic test, we amplified two other gene fragments that are present in S. typhimurium DT104. A multiplex PCR containing primers for targeted sequences resulted in the amplification of predicted size fragments from S. typhimurium DT104 exhibiting the ACSSuT (ampicillin, chloramphenicol, streptomycin, sulphamethoxazole and tetracycline) or ASSuT resistance phenotypes. A minor modification of the multiplex PCR enabled the detection of other related multiresistant Salmonella such as S. typhimurium U302. To augment the detection process, we also designed a fluorogenic PCR assay that can detect the DNA of multiresistant S. typhimurium DT104 in the presence of excess contaminating bacterial DNA. These results provide a method by which multiresistant S. typhimurium DT104, or potentially the next emerging multiresistant Salmonella, can be accurately detected in only 3-4 h.

Drug Resistance, Multiple - genetics

Salmonella typhimurium - genetics

Humans

Repressor Proteins

Rhodamines

Sensitivity and Specificity

Fluoresceins

Drug Resistance, Microbial - genetics

Fluorescent Dyes

Polymerase Chain Reaction - methods

Details

Title: Subtitle: Detection of multiresistant Salmonella typhimurium DT104 using multiplex and fluorogenic PCR
Creators: S A Carlson - Enteric Diseases and Food Safety Research Unit, National Animal Disease Center, Ames, Iowa 50010, USA. scarlson@nadc.ars.usda.gov
L F Bolton
C E Briggs
H S Hurd
V K Sharma
P J Fedorka-Cray
B D Jones
Resource Type: Journal article
Publication Details: Molecular and cellular probes, Vol.13(3), pp.213-222
DOI: 10.1006/mcpr.1999.0240
PMID: 10369747
ISSN: 0890-8508
eISSN: 1096-1194
Language: English
Date published: 06/1999
Academic Unit: Microbiology and Immunology
Record Identifier: 9984083271602771

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