Aim To compare the sequences of the tcdC gene between Clostridioides difficile (C. difficile) strains identified as PCR ribotype 176 and the reference strain C. difficile PCR ribotype 027 and to evaluate the use of the Xpert C. difficile/Epi assay for their differentiation. Methods A total of 45 strains were grown from storage beads. DNA of sufficient quality and quantity for sequencing was extracted from 9 samples. Single consensus sequences of PCR ribotype 176 strains and PCR ribotype 001, PCR ribotype 070 (a control group) were mapped to a reference genome of strain CDI-01 (PCR ribotype 027). Results Four strains (out of seven; 57%) characterized as PCR ribotype 176 had 100% identity of the tcdC gene with the reference strain. The average length of the tcdC gene in these four strains (PCR ribotype 176) was 643 bp, which is 36 bp shorter than the reference genome. Three strains (PCR ribotype 176) had a percentage identity of the tcdC gene in the range of 99.37-100%. Strains 25 (PCR ribotype 001) and 28 (PCR ribotype 070) had a similarity in the range of 95.39-95.63% as a result of different ribotype to the reference strain. Conclusion PCR ribotype 176 strains have almost the same tcdC gene sequence as PCR ribotype 027, resulting in misidentification of this PCR ribotype by the Xpert C. difficile/Epi assay. Information about presumptive positive results based on deletion in the tcdC gene should be treated with caution or disregarded.
Markovska R, Dimitrov G, Gergova R, Boyanova L. Clostridioides difficile, a New "Superbug. Microorganisms. 2023. p. 845.
2.
Liu C, Monaghan T, Yadegar A, Louie T, Kao D. Insights into the Evolving Epidemiology of Clostridioides difficile Infection and Treatment: A Global Perspective. Antibiot Basel Switz. 2023. p. 1141.
3.
Cofini V, Muselli M, Gentile A, Lucarelli M, Lepore R, Micolucci G. Clostridium difficile outbreak: epidemiological surveillance, infection prevention and control. J Prev Med Hyg. 2021.
4.
5.
Edman-Wallér J, Toepfer M, Karp J, Rizzardi K, Jacobsson G, Werner M. Clostridioides difficile outbreak detection: Evaluation by ribotyping and whole-genome sequencing of a surveillance algorithm based on ward-specific cutoffs. Infect Control Hosp Epidemiol. 2023. p. 1948–52.
6.
Kunishima H, Ohge H, Suzuki H, Nakamura A, Matsumoto K, Mikamo H. Japanese Clinical Practice Guidelines for Management of Clostridioides (Clostridium) difficile infection. J Infect Chemother Off J Jpn Soc Chemother. 2022. p. 28.
7.
8.
Camargo T, Junior M, Camargo L, Biotto V, Doi A, Koga P. Clostridioides difficile laboratory diagnostic techniques: a comparative approach of rapid and molecular methods. Arch Microbiol. 2021. p. 1683–90.
9.
Bai Y, Hao Y, Song Z, Chu, Wang J, Y. Evaluation of the Cepheid Xpert C. difficile diagnostic assay: an update meta-analysis. Braz J Microbiol Publ Braz Soc Microbiol. 2021. p. 52.
10.
p. 1937–49.
11.
Oliveira Paiva A, De Jong L, Friggen A, Smits W, Corver J. The C-Terminal Domain of Clostridioides difficile TcdC Is Exposed on the Bacterial Cell Surface. J Bacteriol. 2020. p. 771–90.
12.
Kodori M, Ghalavand Z, Yadegar A, Eslami G, Azimirad M, Krutova M. Molecular characterization of pathogenicity locus (PaLoc) and tcdC genetic diversity among tcdA+B+Clostridioides difficile clinical isolates in Tehran, Iran. Anaerobe. 2020. p. 102294.
13.
Tijerina-Rodríguez L, Villarreal-Treviño L, Morfín-Otero R, Camacho-Ortíz A, Garza-González E. Virulence Factors of Clostridioides (Clostridium) difficile Linked to Recurrent Infections. Can J Infect Dis Med Microbiol J Can Mal Infect Microbiol Medicale. 2019. p. 7127850.
14.
Martínez-Meléndez A, Morfin-Otero R, Villarreal-Treviño L, Baines S, Camacho-Ortíz A, Garza-González E. Molecular epidemiology of predominant and emerging Clostridioides difficile ribotypes. J Microbiol Methods. 2020. p. 105974.
15.
Novakova E, Stefkovicova M, Kopilec M, Novak M, Kotlebova N, Kuijper E. The emergence of Clostridium difficile ribotypes 027 and 176 with a predominance of the Clostridium difficile ribotype 001 recognized in Slovakia following the Eu-ropean standardized Clostridium difficile infection surveillance of 2016. Int J Infect Dis IJID Off Publ Int Soc Infect Dis. 2020. p. 111–5.
16.
Skinner A, Petrella L, Siddiqui F, Sambol S, Gulvik C, Gerding D. Unique Clindamycin-Resistant Clostridioides difficile Strain Related to Fluoroquinolone-Resistant Epidemic BI/RT027 Strain. Emerg Infect Dis. 2020. p. 247–54.
17.
Novakova E, Kotlebova N, Gryndlerova A, Novak M, Vladarova M, Wilcox M. An Outbreak of Clostridium (Clostridioides) difficile Infections within an Acute and Long-Term Care Wards Due to Moxifloxacin-Resistant PCR Ribotype 176 Genotyped as PCR Ribotype 027 by a Commercial Assay. J Clin Med. 2020. p. 3738.
18.
Kameli N, Basode V, Abdulhaq A, Alamoudi M, Zain K, Ghzwani A. Prevalence of toxigenic Clostridium difficile in hospitalized patients in the southwestern province of Saudi Arabia: Confirmation using the GeneXpert analysis. Libyan J Med. 2024. p. 2294571.
19.
Hara T, Suzuki H, Oyanagi T, Koyanagi N, Ushiki A, Kawabata N. Clinical evaluation of a nonpurified direct molecular assay for the detection of Clostridioides difficile toxin genes in stool specimens. PloS One. 2020. p. 234119.
20.
Buddle J, Fagan R. Pathogenicity and virulence of Clostridioides difficile. Virulence. 2023. p. 14.
21.
p. 2150452.
22.
Diniz A, Moura L, Cruz D, Junior O, Figueiredo C, Cunha H, et al. Characterization of the virulence of three novel clade 2 Clostridioides (Clostridium) difficile strains and a twoyear screening in animals and humans in Brazil. PloS One. 2022. p. 273013.
23.
Emele M, Joppe F, Riedel T, Overmann J, Rupnik M, Cooper P. Proteotyping of Clostridioi-des difficile as Alternate Typing Method to Ribotyping Is Able to Distinguish the Ribotypes RT027 and RT176 From Other Ribotypes. Front Microbiol. 2019. p. 2087.
24.
Paulick A, Adamczyk M, Anderson K, Vlachos N, Machado MJ, Mcallister G. Characterization of Clostridioides difficile Isolates Available through the CDC & FDA Antibiotic Resistance Isolate Bank. Microbiol Resour Announc. 2021. p. 1011–20.
25.
Jazmati N, Hain O, Hellmich M, Plum G, Kaasch A. PCR based detection of tcdCΔ117 in Clostridium difficile infection identifies patients at risk for recurrence -A hospital-based prospective observational study. Anaerobe. 2019. p. 39–44.
26.
Okada Y, Yagihara Y, Wakabayashi Y, Igawa G, Saito R, Higurashi Y. Epidemiology and virulence-associated genes of Clostridioides difficile isolates and factors associated with toxin EIA results at a university hospital in Japan. Access Microbiol. 2020. p. 86.
27.
Kachrimanidou M, Baktash A, Metallidis S, Tsachouridou Ο, Netsika F, Dimoglou D. An outbreak of Clostridioides difficile infections due to a 027-like PCR ribotype 181 in a rehabilitation centre: Epidemiological and microbiological characteristics. Anaerobe. 2020. p. 102252.
28.
Riedel T, Neumann-Schaal M, Wittmann J, Schober I, Hofmann J, Lu CW. Characterization of Clostridioides difficile DSM 101085 with A-B-CDT+ Phenotype from a Late Recurrent Colonization. Genome Biol Evol. 2020. p. 566–77.
29.
Novák M, Breznický J, Kompaníková J, Malinovská N, Hudečková H. Impact of hand hygiene knowledge on the hand hygiene compliance. Med Glas (Zenica). 2020. p. 194–9.
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Funding Statement
This work was supported with Comenius University Grant UK 22/2020.
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