Comparison of time-motion analysis of conventional stool culture and the BD MAX™ Enteric Bacterial Panel (EBP)

Mortensen, Joel E.; Ventrola, Cindi; Hanna, Sarah; Walter, Adam

doi:10.1186/s12907-015-0010-8

Cited by 15 publications

(11 citation statements)

References 10 publications

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“…Within the stool culture, direct and enriched culture and EIA specific for identification of STEC account almost one-third of the hands-on time and 59% of the total cost. These data are in agreement with those of others who have reported over 50% of stool culture costs being attributable to STEC-specific testing (10,11). Despite these efforts, the detection of STEC remains low, typically around 1:1,000 specimens in nonoutbreak settings.…”

Section: Resultssupporting

confidence: 92%

“…Because of the increased association of E. coli serotype O:157 with HUS, selective culture using MacConkey agar with sorbitol is also recommended, with confirmation of the O:157 serotype based on latex agglutination. Combined, these efforts to detect STEC account for up to 58% of the total cost associated with the routine workup of stool specimens (10,11). A rapid and inexpensive NAAT can address many of the shortcomings of current methods for detection of STEC by eliminating the need for broth enrichment, selective culture, and serotyping of sorbitol-negative colonies.…”

Section: Resultsmentioning

confidence: 99%

“…Traditional methods for detection of STEC require multiple steps, including direct plating of stool specimens to MacConkey agar with sorbitol (SMAC), serotyping of presumptive serotype O:157 (sorbitolnegative) colonies, and detection of Stx1 and Stx2 using a Shiga toxin enzyme immunoassay (EIA) (6). These approaches are labor-intensive and costly and have poor sensitivity, ranging from 24 to 74% (7)(8)(9)(10)(11). Further, results are not available for 36 to 48 h following specimen culture, which can delay appropriate management of the patient.…”

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confidence: 99%

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Clinical Evaluation and Cost Analysis of Great Basin Shiga Toxin Direct Molecular Assay for Detection of Shiga Toxin-Producing Escherichia coli in Diarrheal Stool Specimens

et al. 2017

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The Shiga Toxin Direct molecular assay (ST Direct) relies on nucleic acid amplification and solid array-based amplicon detection to identify Shiga toxinproducing Escherichia coli (STEC) in preserved stool specimens. Genes encoding Shiga toxin (stx 1 and stx 2 ), as well as the E. coli serotype O:157-specific marker rfbE, are simultaneously detected within 2 h. ST Direct was evaluated using 1,084 prospectively collected preserved stool specimens across five clinical centers. An additional 55 retrospectively collected, frozen specimens were included to increase the number of positive specimens evaluated. Results were compared to results from routine culture and an enzyme immunoassay (EIA) specific for the recovery and identification of STEC. ST Direct was found to be 93.2% sensitive and 99.3% specific for detection of stx 1 and stx 2 and 95.7% sensitive and 99.3% specific for detection of E. coli serotype O:157. All specimens with false-positive results were found to contain stx 1 or stx 2 or were found to be positive for serotype O:157 when analyzed using alternative molecular methods. All 4 false-negative stx 1 or stx 2 results were reported for frozen, retrospectively tested specimens. In all cases, the specimens tested positive for stx by an alternative FDA-cleared nucleic acid amplification test (NAAT) but were negative for stx 1 and stx 2 following nucleic acid sequence analysis. Based on these data, culture and EIA-based methods for detection of STEC are only 33% sensitive compared to molecular tests. A retrospective cost analysis demonstrated 59% of the cost of routine stool culture to be attributable to the identification of STEC. Taken together, these data suggest that ST Direct may provide a costeffective, rapid molecular alternative to routine culture for the identification of STEC in preserved stool specimens.

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Section: Resultssupporting

confidence: 92%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

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Clinical Evaluation and Cost Analysis of Great Basin Shiga Toxin Direct Molecular Assay for Detection of Shiga Toxin-Producing Escherichia coli in Diarrheal Stool Specimens

et al. 2017

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“…Additionally, BDM –EBP PCR results are available 34–47 h sooner that with conventional cultures from rectal swabs [7]. Early detection will aid in epidemiology and managing the spread of these bacterial pathogens.…”

Section: Resultsmentioning

confidence: 99%

Utilizing BD MAX™ Enteric Bacterial Panel to Detect Stool Pathogens from Rectal Swabs

et al. 2017

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BackgroundThe BD MAX™ Enteric Bacterial Panel (BDM-EBP) is designed and FDA-cleared to detect Salmonella, Shigella, Campylobacter, and Shiga toxin genes stx1/2 from stool samples. However, rectal swabs, which are not FDA-cleared for clinical testing with the BDM-EBP, are common specimens received from pediatric patients for enteric pathogen testing. The purpose of this study was to evaluate the ability of the BDM-EBP to detect stool pathogens from rectal swabs.MethodsRoutine cultures, Shiga toxin testing, and molecular testing with BDM-EBP were performed on 272 sequential rectal swabs collected from August 2015 to December 2015. Discrepant test results were resolved using Verigene® Enteric Pathogens Nucleic Acid Test (EP). 36 challenge samples (13 Salmonella spp., 3 Shigella spp., 10 Campylobacter spp., and 10 Shiga toxin positive Escherichia coli) were tested using reference strains (American Type Culture Collection) and previous patient isolates diluted to103-104 cfu/ml in saline then added to Sample Buffer Tube (SBT) with negative stool matrix delivered via a swab. Limit of detection testing was performed by serial 10 fold dilutions in saline then added to SBT with negative stool matrix provided via a swab.ResultsA total of 272 rectal swab specimens were evaluated and 89 were positive by culture and/or MAX EBP. All discrepant results were BDM-EBP positive and culture negative. 21 of 31 (68%) of the apparent false positive BDM-EBP discrepant results resolved as positive with Nanosphere’s Verigene® EP. After resolution of the discordant results, the Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) are as follows for each target: Salmonella (n = 4) 100%, PPA and 100%, NPA; Shigella (n = 79) 100%, PPA and 95.3%, NPA; Campylobacter (n = 4) 100%, PPA and 99.6%, NPA; and Shiga toxin producing organisms (n = 2) 100%, PPA and 100%, NPA. 8.8% of the patient samples did not initially yield a result on the BDM-System. Upon repeat, half of the problematic samples resolved, and 4.4% of the total specimen tested did not yield a result. All organisms in the challenge samples were detected. Limits of detection for BDM-EBP testing of rectal swabs were as follows (in cfu/ml in SBT): Salmonella-1.44 × 102; Shigella-5.10 × 100; Campylobacter-1.51 × 101; and Shiga Toxin-1.13 ×103.ConclusionRectal swabs are acceptable samples for detecting Salmonella, Shigella, Campylobacter, and Shiga toxin using BDM-EBP.

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“…The current gold-standard method for detecting foodborne pathogen in food encompasses enrichment with subsequent plating on selective media, biochemical reactions, and serological tests, which are time-consuming and labor-intensive [26]. Currently,, the official procedure for detection of pathogenic bacteria also used to a cultural method, and this procedure could take from 3 to 5 days for confirmation, which is a disadvantage when the results are needed promptly [27,28]. Hence, faster technologies have been applied to develop rapid and enhance sensitive analytical protocol for the foodborne pathogens.…”

Section: Introductionmentioning

confidence: 99%

Development and Evaluation of a Multiplex PCR for Simultaneous Detection of Five Foodborne Pathogens

Nguyen¹,

Giau²,

Vo³

2016

JARB

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sufficient in specifically and simultaneously detecting as few as 10 CFU/ mL of the five pathogens in artificially inoculated food samples after enrichment for 12 h. Finally, each 25-g sample was mixed with 225mL of SEB medium and incubated at 37 o C for 12-h. Then, each1mL of the culture broth was subjected to the multiplex PCR assay as the schematic representation of detection procedure is presented in Figure 3. The assay is reliable, rapid, specific, and robust. Therefore, it can be another tool for the investigation of microbial contamination in raw food and food products, and will also be useful for identifying the sources of food borne out break

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Comparison of time-motion analysis of conventional stool culture and the BD MAX™ Enteric Bacterial Panel (EBP)

Cited by 15 publications

References 10 publications

Clinical Evaluation and Cost Analysis of Great Basin Shiga Toxin Direct Molecular Assay for Detection of Shiga Toxin-Producing Escherichia coli in Diarrheal Stool Specimens

Clinical Evaluation and Cost Analysis of Great Basin Shiga Toxin Direct Molecular Assay for Detection of Shiga Toxin-Producing Escherichia coli in Diarrheal Stool Specimens

Utilizing BD MAX™ Enteric Bacterial Panel to Detect Stool Pathogens from Rectal Swabs

Development and Evaluation of a Multiplex PCR for Simultaneous Detection of Five Foodborne Pathogens

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