Three-Hour Molecular Detection of Campylobacter, Salmonella, Yersinia, and Shigella Species in Feces with Accuracy as High as That of Culture

Cunningham, Scott A.; Sloan, Lynne M.; Nyre, Lisa M.; Vetter, Emily A.; Mandrekar, Jayawant N.; Patel, Robin

doi:10.1128/jcm.05079-11

Cited by 13 publications

(23 citation statements)

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“…However, broad application remains limited due to their assumed high costs, inhibition caused by fecal constituents (22), and the need for specialized laboratories. Due to the high throughput of stool screening and the number of possible enteric pathogens, implementation of a molecular approach which uses multiplexing of targets is mandatory (9,10,20,25,26,36,43,44,47). For the detection of enteric pathogens it has been proven feasible to use molec-ular methods with improved performance and turnaround time (TAT) (7,25,32 MATERIALS AND METHODS mPCR technical validation.…”

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confidence: 99%

Improved Detection of Five Major Gastrointestinal Pathogens by Use of a Molecular Screening Approach

Boer¹,

et al. 2010

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The detection of bacterial and parasitic gastrointestinal pathogens through culture and microscopy is laborious and time-consuming. We evaluated a molecular screening approach (MSA) for the detection of five major enteric pathogens: Salmonella enterica, Campylobacter jejuni, Giardia lamblia, Shiga toxin-producing Escherichia coli (STEC), and Shigella spp./enteroinvasive E. coli (EIEC), for use in the daily practice of a clinical microbiology laboratory. The MSA consists of prescreening of stool specimens with two real-time multiplex PCR (mPCR) assays, which give results within a single working day, followed by guided culture/microscopy of the positive or mPCR-inhibited samples. In the present 2-year overview, 28,185 stool specimens were included. The MSA was applied to 13,974 stool samples (49.6%), whereas 14,211 samples were tested by conventional methods only (50.4%). The MSA significantly increased the total detection rate compared to that of conventional methods (19.2% versus 6.4%). The detection of all included pathogens, with the exception of S. enterica, significantly improved. MSA detection frequencies were as follows: C. jejuni, 8.1%; G. lamblia, 4.7%; S. enterica, 3.0%; STEC, 1.9%; and Shigella spp./EIEC, 1.4%. The guided culture/microscopy was positive in 76.8%, 58.1%, 88.9%, 16.8%, and 18.1% of mPCR-positive specimens, respectively. Of all mPCRs, only 1.8% was inhibited. Other findings were that detection of mixed infections was increased (0.9% versus 0.02%) and threshold cycle (C T ) values for MSA guided culture/microscopy-positive samples were significantly lower than those for guided culture/microscopy-negative samples. In conclusion, an MSA for detection of gastrointestinal pathogens resulted in markedly improved detection rates and a substantial decrease in time to reporting of (preliminary) results.

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confidence: 99%

Improved Detection of Five Major Gastrointestinal Pathogens by Use of a Molecular Screening Approach

Boer¹,

et al. 2010

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“…The LOD of our assays (10 1 to 10 2 CFU/ml) was below the analytical sensitivity of other reported RT-PCR-based methods [3,4,6,10] and the bacterial burden often reported in symptomatic patients, as measured by culture methods (10 3 to 10 9 CFU/g stool for bacteria). Despite preliminary assays showed consistent results applying the RT-PCRs with DNA extracted from stool specimens, a validation assay in this kind of matrix is required.…”

Section: Discussionmentioning

confidence: 84%

“…and enteroinvasive E. coli (EIEC) (ipaH) [7], enteropathogenic E. coli (EPEC) (eae) [8], enterotoxigenic E. coli (ETEC) (eltA) [4], Shiga toxin-producing E. coli (STEC) (stx 1 and stx 2 ) [9], E. coli O157 (rfbE O157 ) [9], Cronobacter sakazakii (MMS operon) [10], Campylobacter jejuni/C. coli (cadF) [3], Vibrio cholerae (toxR) [4] and Clostridium difficile (tcdB) [4] by real-time PCR.…”

Section: In-house Validation Of Rapid Detection For Bacterial Pathogensmentioning

confidence: 99%

“…When dealing with qualitative analytical methods, the following performance parameters must be considered: sensitivity (limit of detection, LOD), selectivity/specificity (inclusivity/exclusivity) and robustness [2]. Although several polymerase chain reaction (PCR)-based methods to detect bacteria causing diarrhea has been developed elsewhere [3][4][5], none of the protocols have been validated with Argentinean indigenous and regional strains. The objective of this work was to validate 11 inhouse real time-PCRs for the rapid detection of 12 pathogenic bacteria that cause infant diarrhoea.…”

Section: Introductionmentioning

confidence: 99%

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In-House Validation of Rapid Detection PCRs for Bacterial Pathogens Causing Infant Diarrhea

Londero¹,

Ga²,

Brusa³

et al. 2017

J Med Microb Diagn

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“…Rapid PCR assays make use of accelerated cycling for results within three hours. Use of this technique with enterobacterial species showed accuracy comparable to classical culture techniques in a fraction of the time [1,2]. Real-time PCR assays allow higher specificity and shorter assay times than classical PCR techniques as well.…”

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confidence: 94%

Recent Advances in the Biosciences Aid in the Detection and Identification of Bioterrorism Agents

Aune¹,

Burgis²

2011

J Bioterr Biodef

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Three-Hour Molecular Detection of Campylobacter, Salmonella, Yersinia, and Shigella Species in Feces with Accuracy as High as That of Culture

Cited by 13 publications

References 0 publications

Improved Detection of Five Major Gastrointestinal Pathogens by Use of a Molecular Screening Approach

Improved Detection of Five Major Gastrointestinal Pathogens by Use of a Molecular Screening Approach

In-House Validation of Rapid Detection PCRs for Bacterial Pathogens Causing Infant Diarrhea

Recent Advances in the Biosciences Aid in the Detection and Identification of Bioterrorism Agents

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