Clostridioides difficile Ribotype 027 (RT027) Outbreak Investigation Due to the Emergence of Rifampicin Resistance Using Multilocus Variable-Number Tandem Repeat Analysis (MLVA)

Abstract

The aim of this study was Clostridioides difficile outbreak investigation due to the emergence of rifampicin resistant ribotype 027 (RT 027) fecal isolates from patients of Polish tertiary care hospital between X. 2017 and II. 2018 using multilocus variable tandem repeat analysis (MLVA). Materials and Methods: Twenty-nine C. difficile fecal isolates from patients of tertiary care hospital in Southern Poland were ribotyped and analyzed by MLVA. Multiplex PCR (mPCR) for genes encoding GDH (gluD), toxins A (tcdA)/ B (tcdB), 16S rDNA and binary toxin genes (ctdA and ctdB) was performed. The antibiotic susceptibility profile was determined by E-test. Results: The A, B and binary toxins encoding genes were detected in all 29 C. difficile strains which were sensitive to metronidazole, vancomycin and were resistant to erythromycin, clindamycin, and moxifloxacin; resistance to imipenem demonstrated 97%, to rifampicin -45% isolates. C. difficile strains could be grouped by MLVA into 5 distinct clusters, and the largest cluster II contains 16 strains. The comparison of rifampicin GM MIC of cluster II (n=16 strains) with all others (n=13) showed that strains from clusters I, III, IV and V possessed significantly (p <0.005) higher GM MIC and were more resistant to rifampicin. Conclusion: MLVA analysis proved transmission and recognized outbreak due to multidrugresistant RT 027 C. difficile among patients of tertiary care hospital in Southern Poland. The reason for this is probably the widespread occurrence of spores in the hospital environment, which includes, among others, neglect of hygienic procedures and epidemic supervision. High resistance to imipenem (97%) and to rifampicin (45%) among C. difficile RT 027 Silesian isolates is threatening and requires further studies to elucidate this phenomenon.

High Prevalence of Clostridioides difficile Ribotype 176 in the University Hospital in Kosice

“…Our results indicate that the proportions of CD ribotypes are probably highly connected to the specific hospital departments, and the nosocomial transmission of CD strains from one department to another is common. Similar findings are stated by several studies [ 46 , 51 , 52 ]. Most of these outbreaks were recorded between August and December, during the second wave of the COVID-19 pandemic.…”

Outbreak of <i>Clostridioides difficile</i> infection in Silesian district hospital

“…W Y N I K I: Wśród pacjentów dominowały kobiety (4/5). Wszystkie szczepy C. difficile (6) należały do hiperepidemicznego rybotypu 027, we wszystkich szczepach wykazano obecność badanych genów oraz wykryto oporność na: moksyfloksacynę, erytromycynę, klindamycynę, ryfampicynę, imipenem i chloramfenikol. Clostridioides difficile is an anaerobic, Gram-positive spore-forming bacillus.…”

“…An increase in severe CDI caused by the C. difficile ribotype (RT) 027 strain was reported in Canada and the USA in 2000-2003. During the next 10 years, the dominance of the hyperepidemic strain was noted in European countries, including Poland and on other continents [6,7]. The frequent occurrence of this strain has been observed especially in southern Poland and also in Western Australia, South Korea, Hong Kong and Costa Rica [8].…”

“…The fecal samples were examined in accordance with a two-stage algorithm: first, detecting the C. difficile somatic antigenglutamate dehydrogenase (GDH) and then toxins A/B -(TechLab, Blacksburg, USA) [13]. Next, (in the case of one or both positive results) the stool samples were cultured on chromID C. difficile and CLO plates (bioMérieux, Marcy L'Etoile, France), incubated at 37°C under anaerobic conditions (Whitley A35 Workstation, UK) for 48 h. Colonies with characteristic morphology (CDIFFblack colonies, CLOgray, jagged with yellow-green fluorescence under UV light, and horse odor) were isolated for further biochemical identification (VITEK 2 Compact System, bioMérieux, Marcy L'Etoile, France), antibiotic susceptibility testing and ribotyping [6]. The genes encoding C. difficile toxins and GDH were detected by performing the multiplex polymerase chain reaction (mPCR) according to Stubbs, and using the HotStarTaq Plus PCR Master Mix Kit (Qiagen, Germany) [14].…”

Clostridium perfringens Spores in Urology Hospital

“…The GeneMATRIX DNA Purification Kit by DNA Gdansk, PL, was used for the same purpose. The genes encoding A, B, and binary toxins in C. difficile were detected by multiplex PCR (mPCR), as previously described (12). Moreover, mPCR was used to detect the presence of toxin alpha-cpa, toxin beta-cbp, enterotoxin-cpe, iota toxin-cpiA, and epsilon toxin-etxin C. perfringens, as described by Meer and Songer (13).…”