Rapid Simultaneous Molecular Stool-Based Detection of Toxigenic Clostridioides Difficile by Quantitative Taqman Real-Time Pcr Assay

Rapid Simultaneous Molecular Stool-Based Detection of Toxigenic Clostridioides Difficile by Quantitative Taqman Real-Time Pcr Assay Publisher Pubmed

Kouhsari E^{1, 2} ; Douraghi M³ ; Barati M⁴ ; Yaseri HF⁵ ; Talebi M¹ ; Abbasian S¹ ; Moqarabzadeh V⁶ ; Amirmozafari N¹

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1. Dept of Microbiology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
2. Laboratory Sciences Research Center, Golestan University of Medical Sciences, Gorgan, Iran
3. Division of Microbiology, Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
4. Dept of Biotechnology, School of Allied Medicine, Iran University of Medical Science, Tehran, Iran
5. Research Center for Gastroenterology and Liver Disease, Firouzgar Hospital, Iran University of Medical Sciences, Tehran, Iran
6. Dept of Biostatistics, School of Health, Mazandaran University of Medical Sciences, Sari, Iran

Source: Clinical Laboratory Published:2019

Abstract

Background: Clostridioides difficile is a major cause of nosocomial infectious diarrhea in hospitalized patients throughout the world. Methods: A multiplex real-time PCR assay was developed and evaluated in comparison with toxigenic culture (TC) (as gold standard method) for direct detection of toxigenic C. difficile in fecal specimens. The multiplex real-time PCR assay simultaneously detected glutamate dehydrogenase (gluD), toxin A (tcdA), toxin B (tcdB), and binary toxin (cdtB) genes in stool samples. Results: The results of multiplex real-time PCR were compared to those of the TC method in 250 patients suspected of C. difficile infection. The prevalence of positive TC was 13.6%. Forty-two stool samples (16.8%) were determined to be gluD+ using multiplex real-time PCR. These included 35 (83.3%) toxigenic (32 tcdA+, tcdB+ and three tcdB+) and 7 (20.0%) were cdtB+. The multiplex real-time PCR assay had a sensitivity of 91.45%, specificity of 99.54%, and positive and negative predictive values of 97% and 98.6%, respectively, compared to the TC method for diagnosis of C. difficile. The analytical sensitivity of the multiplex real-time PCR assay was estimated to be 102 CFU/g of stools and 0.0200 pg of genomic DNA from culture. The analytical specificity was determined to be 100% by using enteric and non-C. difficile standard bacterial strains. Conclusions: The molecular method developed in the study was rapid, sensitive, and specific for detection of toxigenic C. difficile. It is applicable to be performed in clinical laboratories and correlated well with the results obtained by TC. © 2019 Verlag Klinisches Labor GmbH. All rights reserved.

2. The Emergence of Metronidazole and Vancomycin Reduced Susceptibility in Clostridium Difficile Isolates in Iran, Journal of Global Antimicrobial Resistance (2019)

3. 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, Archives of Pediatric Infectious Diseases (2017)

4. Pathogenicity Locus Determinants and Toxinotyping of Clostridioides Difficile Isolates Recovered From Iranian Patients, New Microbes and New Infections (2018)

5. Isolation and Characterization of a Multidrug-Resistant Clostridioides Difficile Toxinotype V From Municipal Wastewater Treatment Plant, Journal of Environmental Health Science and Engineering (2020)

6. A Two-Step Approach for Diagnosing Glutamate Dehydrogenase Genes by Conventional Polymerase Chain Reaction From Clostridium Difficile Isolates, Middle East Journal of Digestive Diseases (2019)

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Masoumeh Douraghi (4)

Mehdi Saberi Firoozi (1)

Style	Citing Format
MLA	Kouhsari E, et al.. "Rapid Simultaneous Molecular Stool-Based Detection of Toxigenic Clostridioides Difficile by Quantitative Taqman Real-Time Pcr Assay." Clinical Laboratory, vol. 65, no. 4, 2019, pp. 461-469.
APA	Kouhsari E, Douraghi M, Barati M, Yaseri HF, Talebi M, Abbasian S, Moqarabzadeh V, Amirmozafari N (2019). Rapid Simultaneous Molecular Stool-Based Detection of Toxigenic Clostridioides Difficile by Quantitative Taqman Real-Time Pcr Assay. Clinical Laboratory, 65(4), 461-469.
Chicago	Kouhsari E, Douraghi M, Barati M, Yaseri HF, Talebi M, Abbasian S, Moqarabzadeh V, Amirmozafari N. "Rapid Simultaneous Molecular Stool-Based Detection of Toxigenic Clostridioides Difficile by Quantitative Taqman Real-Time Pcr Assay." Clinical Laboratory 65, no. 4 (2019): 461-469.
Harvard	Kouhsari E et al. (2019) 'Rapid Simultaneous Molecular Stool-Based Detection of Toxigenic Clostridioides Difficile by Quantitative Taqman Real-Time Pcr Assay', Clinical Laboratory, 65(4), pp. 461-469.
Vancouver	Kouhsari E, Douraghi M, Barati M, Yaseri HF, Talebi M, Abbasian S, et al.. Rapid Simultaneous Molecular Stool-Based Detection of Toxigenic Clostridioides Difficile by Quantitative Taqman Real-Time Pcr Assay. Clinical Laboratory. 2019;65(4):461-469.
BibTex	@article{ author = {Kouhsari E and Douraghi M and Barati M and Yaseri HF and Talebi M and Abbasian S and Moqarabzadeh V and Amirmozafari N}, title = {Rapid Simultaneous Molecular Stool-Based Detection of Toxigenic Clostridioides Difficile by Quantitative Taqman Real-Time Pcr Assay}, journal = {Clinical Laboratory}, volume = {65}, number = {4}, pages = {461-469}, year = {2019} }
RIS	TY - JOUR AU - Kouhsari E AU - Douraghi M AU - Barati M AU - Yaseri HF AU - Talebi M AU - Abbasian S AU - Moqarabzadeh V AU - Amirmozafari N TI - Rapid Simultaneous Molecular Stool-Based Detection of Toxigenic Clostridioides Difficile by Quantitative Taqman Real-Time Pcr Assay JO - Clinical Laboratory VL - 65 IS - 4 SP - 461 EP - 469 PY - 2019 ER -

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