Comparison of the Detection Limits of the Culture and PCR Methods for the Detection of Clostridium difficile, Clostridium perfringens, Campylobacter jejuni, and Yersinia enterocolitica in Human Stool

Ganji, Leila; Azimirad, Masoumeh; Farzi, Nastaran; Alebouyeh, Masoud; Shirazi, Mohammad Hassan; Eshraghi, Seyyed Saeed; Mirshafiey, Abbas; Daryani, Naser Ebrahimi; Zali, Mohammad Reza

doi:10.5812/pedinfect.38888

Cited by 8 publications

(5 citation statements)

References 42 publications

(40 reference statements)

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“…difficile in several culture-negative cases [ 21 ]. In a more recent analysis involving spiked stool samples, the LOD of traditional PCR analyzed by gel electrophoresis was 10-fold higher than culture [ 22 ]. Overall, we found the LOD for qPCR was 3.2-fold lower compared to culture for environmental detection of C .…”

Section: Discussionmentioning

confidence: 99%

Comparison of qPCR versus culture for the detection and quantification of Clostridium difficile environmental contamination

et al. 2018

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Contaminated surfaces serve as an important reservoir for Clostridium difficile transmission. Current strategies to detect environmental contamination of C. difficile rely heavily on culture, and often only indicate presence versus absence of spores. The goal of this study was to compare quantitative PCR (qPCR) to culture for the detection and quantification of C. difficile from inert surfaces. First, we compared the limit of detection (LOD) of a 16S rRNA gene and toxin B gene qPCR assay for detection of C. difficile in solution. Second, we compared the LODs of 16S rRNA gene qPCR versus culture for detection of C. difficile from surfaces. Solution experiments were performed by direct seeding of spores into neutralizing broth, whereas surface experiments involved seeding of spores onto plastic test surfaces, and recovery using sponge swabs. Both experiments were conducted using spores expressing short (NAP1) and long (NAP4) hair lengths. Combining data from both strains, the overall LOD for C. difficile cells in solution was 1.4 cells for 16S rRNA gene and 23.6 cells for toxin B gene qPCR (p<0.001). The overall LOD for C. difficile cells from surfaces was 17.1 cells for 16S rRNA gene qPCR and 54.5 cells for culture (p = 0.05), and was not statistically different between strains for each method (p = 0.52). Overall, the proportion of C. difficile cells recovered from surfaces was good when detected by 16S rRNA gene qPCR and culture (qPCR: 76%, culture: 67%, p = 0.36), but, 16S rRNA gene qPCR was capable of detecting lower levels of surface contamination. Future work attempting to measure the presence of C. difficile on environmental surfaces should consider using qPCR.

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Section: Discussionmentioning

confidence: 99%

Comparison of qPCR versus culture for the detection and quantification of Clostridium difficile environmental contamination

et al. 2018

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“…As a comparison, EIA against GDH or EIA against toxin A/B can reach 10 3 CFU/g (feces) [58] . The LOD of PCR assays varies from 10 1 to 10 4 CFU/mL (CFU/g) [59–61] . Overall, RFD‐CD2 based assays displayed very comparable LODs with these two detection methods.…”

Section: Resultsmentioning

confidence: 96%

“…[58] The LOD of PCR assays varies from 10 1 to 10 4 CFU/mL (CFU/g). [59][60][61] Overall, RFD-CD2 based assays displayed very comparable LODs with these two detection methods. The culturing methods, however, can reach an LOD of 10 1 to 10 2 CFU/mL depending on the recovery method.…”

Section: Detection Sensitivity Of Rfd-cd2 As a Fluorescent Sensormentioning

confidence: 99%

In Vitro Selection and Characterization of a DNAzyme Probe for Diverse Pathogenic Strains of Clostridium difficile

Qian

Chang

et al. 2023

Chemistry A European J

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Clostridium difficile frequently causes an infectious disease known as Clostridium difficile infection (CDI), and there is an urgent need for the development of more effective rapid diagnostic tests for CDI. Previously we have developed an RNA-cleaving fluorogenic DNAzyme (RFD) probe, named RFD-CD1, that is capable of detecting a specific strain of C. difficile but is too specific to recognize other pathogenic C. difficile strains. To overcome this issue, herein we report RFD-CD2, another RFD that is not only highly specific to C. difficile but also capable of recognizing diverse pathogenic C. difficile strains. Extensive sequence and structure characterization establishes a pseudoknot structure and a significantly minimized sequence for RFD-CD2. As a fluorescent sensor, RFD-CD2 can detect C. difficile at a concentration as low as 100 CFU/mL, thus making this DNAzyme an attractive molecular probe for rapid diagnosis of CDI caused by diverse strains of C. difficile.

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“…Bacterial cell culture is currently the gold standard method for identifying contamination; however it often fails to detect low concentrations of bacteria or spores, resulting in false negative outcomes [ 26 ]. Also, standard culture assays require several days for analysis.…”

Section: Discussionmentioning

confidence: 99%

Ultrasensitive qPCR platform for rapid detection of bacterial contamination of raw biological samples at the point of care

Garzarelli

Chiriaco

Cereda

et al. 2023

Heliyon

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Comparison of the Detection Limits of the Culture and PCR Methods for the Detection of Clostridium difficile, Clostridium perfringens, Campylobacter jejuni, and Yersinia enterocolitica in Human Stool

Cited by 8 publications

References 42 publications

Comparison of qPCR versus culture for the detection and quantification of Clostridium difficile environmental contamination

Comparison of qPCR versus culture for the detection and quantification of Clostridium difficile environmental contamination

In Vitro Selection and Characterization of a DNAzyme Probe for Diverse Pathogenic Strains of Clostridium difficile

Ultrasensitive qPCR platform for rapid detection of bacterial contamination of raw biological samples at the point of care

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