Infecciones por Clostridioides difficile en pediatría. La visión del laboratorio a partir del diagnóstico microbiológico / Clostridioides difficile infections in pediatrics. The view of the laboratory based on microbiological diagnosis

La infección por Clostridioides difficile (ICD) es la principal responsable de diarreas nosocomiales en adultos. En los últimos años se registró un aumento en la incidencia de la ICD en la población adulta que, en cambio, no fue bien caracterizado en pediatría. El objetivo de este trabajo es analizar los datos resultantes del diagnóstico microbiológico de ICD en el Hospital de Pediatría "Prof. Dr. Juan P. Garrahan". Materiales y métodos: se realizó un estudio retrospectivo observacional descriptivo que abarcó desde el 01/01/2018 hasta el 31/12/2021. El diagnóstico se realizó mediante enzimoinmunoensayo para glutamato deshidrogenasa (GDH) y toxinas en materia fecal (MF). Cuando sólo se detectó GDH, se realizó un cultivo toxigénico (CT) de la MF para la detección de toxinas in vitro. Se registraron: edad, sexo y procedencia de los pacientes y recurrencias de las ICD. Se efectuaron estudios de sensibilidad de 387 cepas de C. difficile a metronidazol (MTZ) y vancomicina (VAN). Resultados: en 6632 muestras (1764 pacientes) se registraron 649 estudios positivos (9,8%) (139 pacientes), la mayoría correspondieron a pacientes internados en áreas no críticas. Edad promedio: 7 años (7 ± 4,7). Sexo: 55% masculino. Recurrencias: 62 (45%). Positivos detectados mediante CT: 43%. Sensibilidad antibiótica: 100% a MTZ y 99,7% a VAN. Conclusión: Nuestra población presenta un bajo porcentaje de positividad. Se destaca el rendimiento del CT que permitió el diagnóstico de más de un tercio de los casos. MTZ y VANCO tuvieron excelente actividad in vitro frente a C. difficile (AU)

Clostridioides difficile infection (CDI) is the main cause of nosocomial diarrhea in adults. In recent years there has been an increase in the incidence of CDI in the adult population; however, CDI has not been well characterized in pediatrics. The aim of this study was to analyze the data resulting from the microbiological diagnosis of CDI at Hospital de Pediatría Prof. Dr. Juan P. Garrahan. Materials and methods: a retrospective, observational and descriptive study was conducted from 01/01/2018 to 12/31/2021. Diagnosis was made using enzyme immunoassay for glutamate dehydrogenase (GDH) and toxins in stools. When only GDH was detected, toxigenic culture (TC) of stools was performed for in vitro toxin detection. The age, sex and origin of patients and CDI recurrences were recorded. Sensitivity studies of 387 strains of C. difficile to metronidazole (MTZ) and vancomycin (VAN) were performed. Results: In 6,632 samples (1,764 patients), 649 positive results (9.8%) were recorded (139 patients), most of which corresponded to patients hospitalized in noncritical areas. Mean age: 7 years (7 ± 4.7). Sex: 55% male. Recurrences: 62 (45%). TC-positive results: 43%. Antibiotic sensitivity: 100% to MTZ and 99.7% to VAN. Conclusion: A low percentage of positivity was found in our population. The performance of TC was outstanding, allowing for the diagnosis of more than one third of the cases. MTZ and VANCO had excellent in vitro activity against C. difficile (AU)

Medicamentos antidepressivos estão associados a um risco aumentado de infecção hospitalar por Clostridioides difficile: um estudo populacional / Antidepressant medications are associated with increased risk of hospital-acquired clostridioides difficile infection: a population-based study

ABSTRACT Background: During the past decade, Clostridioides difficile infection (CDI) has become the most common cause of antibiotic-associated diarrhea. Several risk factors have been implicated. Scattered evidence about the association of CDI with antidepressant medications use exists in the literature so far. Therefore, we aim to investigate whether the risk of developing CDI is increased in hospitalized patients using antidepressant medications. Methods: Patients who were hospitalized were included in our cohort. We excluded individuals aged less than 18 years. A multivariate regression analysis was performed to calculate the risk of CDI accounting for potential confounders. Results: The risk of CDI in hospitalized patients was increased in individuals diagnosed with inflammatory bowel disease (OR: 4.44; 95%CI: 4.35-4.52), and in patients using clindamycin (OR: 1.55; 95%CI: 1.53-1.57), beta-lactam antibiotics (OR: 1.62; 95%CI: 1.60-1.64), PPI (OR: 3.27; 95%CI: 3.23-3.30), trazodone (OR: 1.31; 95%CI: 1.29-1.33), nortriptyline (OR: 1.25; 95%CI: 1.21-1.28), and mirtazapine (OR: 2.50; 95%CI: 2.46-2.54). After controlling for covariates, the risk of CDI was not increased in patients who were taking fluoxetine (OR: 0.94; 95%CI: 0.92-0.96). Conclusion: In contrary to fluoxetine; mirtazapine, nortriptyline, and trazodone were associated with increased risk of CDI in hospitalized patients.

RESUMO Contexto: Na última década, a infecção por Clostridioides difficile (ICD) tornou-se a causa mais comum de diarreia associada a antibióticos. Vários fatores de risco foram implicados. Existem evidências dispersas na literatura sobre a associação da ICD com o uso de medicamentos antidepressivos. Portanto, pretendemos investigar se o risco de desenvolver infecção adquirida na comunidade por Clostridioides difficile aumenta em pacientes que usam medicamentos antidepressivos. Métodos: Pacientes que foram hospitalizados foram incluídos em nossa coorte. Indivíduos com menos de 18 anos foram excluídos. Uma análise de regressão multivariada foi realizada para calcular o risco de ICD, considerando possíveis confusões. Resultados: O risco de ICD em pacientes hospitalizados foi maior em indivíduos diagnosticados com doença inflamatória intestinal (OR: 4,44; IC95%: 4,35-4,52) e em pacientes que usavam clindamicina (OR: 1,55; IC95%: 1,53-1,57), antibióticos beta-lactâmicos (OR: 1,62; IC95%: 1,60-1,64), PPI (OR: 3,27; IC95%: 3,23-3,30), trazodona (OR: 1,31; IC95%: 1,29-1,33), nortriptilina (OR: 1,25; IC95%: 1,21-1,28) e mirtazapina (OR: 2,50; IC95%: 2,46-2,54). Depois de controlar as covariáveis, o risco de ICD não aumentou em pacientes que estavam tomando fluoxetina (OR: 0,94; IC95%: 0,92-0,96). Conclusão: Em contrário à fluoxetina; mirtazapina, nortriptilina e trazodona foram associados a um risco aumentado de ICD em pacientes hospitalizados.

Characterization of a Clostridioides difficile ST-293 isolate from a recurrent infection in Argentina

Resumen Clostridioides difficile es un patógeno esporulado oportunista responsable de diarrea asociada a antibióticos en humanos. C. difficile produce 2 toxinas principales: TcdAy TcdB, además de la toxina binaria (CDT), también asociada a la virulencia. Este estudio buscó caracterizar el aislamiento ALCD3, involucrado en un episodio de recurrencia de una infección nosocomial. La caracterización molecular mostró que dicho aislamiento pertenece al toxinotipo 0/v y el análisis por MLST demostró un perfil alélico adk:91, atpA:1, dxr:2, glyA: 1, recA:27, sodA: 1 y tpi:1, lo cual corresponde al ST293 (MLST clado 1). Durante el crecimiento, el aislamiento ALCD3 mostró un incremento temprano de la tasa de esporulación y valores máximos de formas termorresistentes luego de 2 días de incubación. Tanto la cinética de esporulación como la producción de formas termorresistentes fueron más rápidas en el aislamiento ALCD3 que en la cepa de referencia VPI 10463. La germinación en presencia del germinante natural taurocolato fue más rápida en el aislamiento ALCD3 que en la cepa VPI 10463, lo que indica que aquel comienza la hidrólisis del córtex antes. También, el co-germinante glicina indujo una rápida liberación de ácido dipicolínico en ALCD3. Estos hallazgos indican que el aislamiento ALCD3 es particularmente eficiente en la esporulación y en la germinación. El presente trabajo representa el primer informe de la circulación de C. difficile ST293 en Argentina. La habilidad del aislamiento ALCD3 para producir toxinas y su alta capacidad de esporulación/germinación son características claves compatibles con un alto potencial de diseminación e inducción de infecciones recurrentes.

Abstract Clostridioides difficile is an opportunistic spore-forming pathogen responsible for antibiotic-associated diarrhea in humans. C. difficile produces two main toxins: TcdA and TcdB as well as a third toxin named binary toxin (CDT) that is also involved in virulence. The present study aimed at characterizing the C. difficile isolate ALCD3 involved in a relapse episode of nosocomial infection. Molecular characterization showed that isolate ALCD3 belongs to tox-inotype 0/v and the MLST analysis demonstrated allelic profile adk:91, atpA:1, dxr:2, glyA: 1, recA:27, sodA: 1 and tpi:1 which corresponds to ST293 (MLST clade: 1). During growth, isolate ALCD3 showed an early increase in the sporulation ratio as well as maximal values of heat resis-tant forms after 2 days of incubation. Both sporulation kinetics and production of heat resistant forms were faster for isolate ALCD3 than for the reference strain VPI 10463. Germination in the presence of the natural germinant taurocholate was faster for isolate ALCD3 than for strain VPI 10463, which indicates that isolate ALCD3 starts cortex hydrolysis earlier than strain VPI 10463. Furthermore, the co-germinant glycine, induces rapid release of dipicolinic acid (DPA) in isolate ALCD3. These findings indicate that isolate ALCD3 is particularly efficient in both sporulation and germination. The present work represents the first report of the circulation of C. difficile ST293 in Argentina. The ability of isolate ALCD3 to produce toxins and its high sporulation/germination capacity are key features compatible with a microorganism with high dissemination potential and the possibility of inducing recurrent infections.

Trasplante de microbiota fecal como alternativa de tratamiento por infecciones de clostridoides difficile / Fecal microbiota transplantation as a treatment alternative for clostridoides difficile infections / Transplante de microbiota fecal como tratamento alternativo para infecções clostridioides difficile

La infección por Clostridioides difficile es una amenaza para la salud pública, está asociada a la atención médica, cuya complicación más frecuente es la infección recurrente, con tasas de hasta el 60% después del tercer episodio. Las opciones de tratamiento para la recurrencia de esta infección son limitadas. Una gran paradoja es tratar una infección asociada a antibióticos con más antibióticos, por ello, la piedra angular en el manejo de esta infección es la restauración de la microbiota intestinal mediante el trasplante de microbiota fecal. Objetivo. Determinar la eficacia y seguridad del trasplante de microbiota fecal para el tratamiento de la infección recurrente por Clostridioides difficile. Metodología. Se realizó una revisión bibliográfica narrativa de la literatura científica en las bases de datos PubMed y Cochrane Library empleando los Descriptores en Ciencias de la Salud (DeCS) y Medical Subject Headings (MeSH), junto con los operadores booleanos "AND/Y", "OR/O"; donde se recopilaron los estudios que cumplieron con los criterios de inclusión. Conclusión. Se concluyó que el trasplante de microbiota fecal en la infección recurrente por Clostridioides difficile es un tratamiento eficaz y seguro, con eventos adversos mínimos, aunque la seguridad a largo plazo no está bien establecida.

Clostridioides difficile infection is a public health threat, is associated with health care, the most common complication of which is recurrent infection, with rates of up to 60% after the third episode. Treatment options for recurrence of this infection are limited. A great paradox is to treat an antibiotic-associated infection with more antibiotics; therefore, the cornerstone in the management of this infection is the restoration of the intestinal microbiota by fecal microbiota transplantation. Objective. To determine the efficacy and safety of fecal microbiota transplantation for the treatment of recurrent Clostridioides difficile infection. Methodology. A narrative bibliographic review of the scientific literature was carried out in the PubMed and Cochrane Library databases using the Health Sciences Descriptors (DeCS) and Medical Subject Headings (MeSH), together with the Boolean operators "AND/Y", "OR/O"; where the studies that met the inclusion criteria were collected. Conclusion. It was concluded that fecal microbiota transplantation in recurrent Clostridioides difficile infection is an effective and safe treatment, with minimal adverse events, although long-term safety is not well established.

A infecção por Clostridioides difficile é uma ameaça à saúde pública associada ao cuidado com a saúde, cuja complicação mais comum é a infecção recorrente, com taxas de até 60% após o terceiro episódio. As opções de tratamento para infecções recorrentes são limitadas. Um grande paradoxo é tratar uma infecção associada a antibióticos com mais antibióticos, portanto, a pedra fundamental no manejo desta infecção é a restauração da microbiota intestinal através do transplante da microbiota fecal. Objetivo. Determinar a eficácia e segurança do transplante de microbiota fecal para o tratamento de infecções recorrentes por Clostridioides difficile. Metodologia. Uma revisão bibliográfica narrativa da literatura científica foi realizada nas bases de dados da Biblioteca PubMed e Cochrane utilizando os Descritores de Ciências da Saúde (DeCS) e os Títulos de Assuntos Médicos (MeSH), juntamente com os operadores booleanos "AND/Y", "OR/O"; onde foram compilados os estudos que preenchiam os critérios de inclusão. Conclusão. Concluiu-se que o transplante de microbiota fecal em infecção recorrente por Clostridioides difficile é um tratamento eficaz e seguro com o mínimo de eventos adversos, embora a segurança a longo prazo não esteja bem estabelecida.

Clostridioides difficile: Characterization of the circulating toxinotypes in an Argentinean public hospital / Clostridioides difficile: caracterización de los toxinotipos circulantes en un hospital público de Argentina

Abstract Clostridioides difficile is a spore-forming anaerobe microorganism associated to nosocomial diarrhea. Its virulence is mainly associated with TcdA and TcdB toxins, encoded by their respective tcdA and tcdB genes. These genes are part of the pathogenicity locus (PaLoc). Our aim was to characterize relevant C. difficile toxinotypes circulating in the hospital setting. The tcdA and tcdB genes were amplified and digested with different restriction enzymes: EcoRI for tcdA; HincII and AccI for tcdB. In addition, the presence of the cdtB (binary toxin) gene, TcdA and TcdB toxins by dot blot and the cytotoxic effect of culture supernatants on Vero cells, were evaluated. Altogether, these studies revealed three different circulating toxinotypes according to Rupnik's classification: 0, I and VIII, being the latter the most prevalent one. Even though more studies are certainly necessary (e.g. sequencing analysis), it is worth noting that the occurrence of toxinotype I could be related to the introduction of bacteria from different geographical origins. The multivariate analysis conducted on the laboratory values of individuals infected with the most prevalent toxinotype (VIII) showed that the isolates associated with fatal outcomes (GCD13, GCD14 and GCD22) are located in regions of the biplots related to altered laboratory values at admission. In other patients, although laboratory values at admission were not correlated, levels of urea, creatinine and white blood cells were positively correlated after the infection was diagnosed. Our study reveals the circulation of different toxinotypes of C. difficile strains in this public hospital. The variety of toxinotypes can arise from pre-existing microorganisms as well as through the introduction of bacteria from other geographical regions. The existence of microorganisms with different pathogenic potential is relevant for the control, follow-up, and treatment of the infections.

Resumen Clostridioides difficile es un anaerobio esporulado que se asocia con episodios de diarreas hospitalarias. Su virulencia se encuentra vinculada, principalmente, a las toxinas TcdA y TcdB, codificadas por sus respectivos genes, tcdA y tcdB, que son parte de un locus de patogenicidad (PaLoc). Nuestro objetivo fue caracterizar los toxinotipos de C. difficile circulantes en un hospital público. Los genes tcdA y tcdB fueron amplificados y digeridos con diferentes enzimas de restricción: EcoRI para tcdA; HincII y AccI para tcdB. Además, se evaluó la presencia de cdtB (gen de la toxina binaria B) y de las toxinas A y B (por dot blot), así como el efecto citotóxico de sobrenadantes de cultivo sobre células Vero. En conjunto, estos estudios revelaron tres toxinotipos circulantes según la clasificación de Rupnik: 0, I y VIII; el más prevalente fue el último. Aunque son necesarios más estudios (ej., secuenciación), es interesante notar que la presencia del toxinotipo I podría estar relacionada con la introducción de bacterias de diferente origen geográfico. En los pacientes infectados con el toxinotipo VIII, el análisis multivariante de los resultados de laboratorio mostró que los aislamientos asociados a decesos (GCD13, GCD14 y GCD22) estaban situados en regiones de los biplots relacionados con valores de laboratorio alterados al momento de la internación. En los otros pacientes, aunque no se observó correlación entre los valores de laboratorio al momento de la internación y la evolución clínica, los niveles de urea, creatinina y recuento de glóbulos blancos estuvieron correlacionados positivamente entre sí una vez diagnosticada la infección. Nuestro estudio revela la circulación de diferentes toxinotipos de C. difficile en un mismo hospital público. La variedad de toxinotipos puede originarse a partir de microorganismos preexistentes en la región, así como también por la introducción de bacterias provenientes de otras regiones geográficas. La existencia de microorganismos con diferente potencial patogénico es relevante para el control, el seguimiento y el tratamiento de las infecciones.

Status and challenge of vaccine research for Clostridiodes difficile in clinical trials / 中国生物制品学杂志

@#Clostridiodes difficile(C.difficile)is the most common causative agent of antibiotic-associated diarrhea(ADD)in the world. In recent years,with the emergence of highly resistant and virulent strains,the outbreaks of C.difficile infection have occurred around the world. The incidence,recurrence and mortality of C.difficile infection are on the rise worldwide,and bring great challenges to clinical treatment. Pathogenic strains mainly produce two homologous glycosylation toxins A and B,which can cause symptoms ranging from diarrhea to highly lethal toxic megacolon. In view of the malignant consequences caused by C.difficile infection,disease prevention is still an important way worth exploring. Until now there is no approved vaccine against C.difficile. Therefore,this review assessed the status and challenge of clinical trials of vaccine research for C.difficile.

Prokaryotic expression,purification and activity of CamA protein with methylase activity from Clostridiodies difficile / 中国生物制品学杂志

@#Objective To express CamA(Clostridiodies difficile adenine methltransferase A,CamA)protein with methylase activity in prokaryotic expression system.Methods The gene sequence of CamA protein of the standard strain of C.difficile1870 was amplified by PCR,cloned into plasmid pGEX-4T-1-MBP,transformed into E.coli HB101 and induced by 1 mmol/L IPTG to express the recombinant target protein. After purification with Amylose Resin,CamA protein was digested by tobacco etch virus(TEV)protease,and verified for its methylase activity using MTase-Glo~(TM)Methyltransferase Assay in vitro.Results PCR sequencing showed that the cloned CamA gene sequence was correct,in which no base mutation occurred. The recombinant expression plasmid pGEX-4T-1-MBP-CamA was digested by EcoRⅠand BamHⅠ,which showed that 1 731 bp of CamA gene was connected to the vector plasmid. The relative molecular mass of the recombinant protein was about 108 800(with 1 MBP tag),and the purity was over 90% after purified with Amylose Resin. Under the condition of20 μmol/L DNA and 20 μmol/L SAM at room temperature for 30 min,0. 5 μg CamA produced SAH at a concentration of about 101. 60 nmol/L and the enzyme activity was(0. 339 ± 0. 027)U/mg.Conclusion The CamA protein of C.difficile has been successfully expressed and purified with methylase activity,which lays a foundation of further study on the function of CamA and its role in the occurrence,development and treatment of C.difficile infection.

Comparison of biological characteristics of major epidemic strains of Clostridium difficile from nosocomial infectious diarrhea / 中华检验医学杂志

Objective:This work aims to investigate the virulence features, spore formation and the resistance mechanisms of major sequence types (STs) of clinical Clostridium difficile isolates from nosocomial infectious diarrhea. Methods:Clostridium difficile isolates were prospectively collected from 816 loose stool samples of in patients with antibiotic associated diarrhea at the Beijing Friendship Hospital of Capital Medical University from September 2017 to September 2019. The main ST types ST81 (26 strains), ST8 (15 strains) and ST42 (14 strains) of C. difficile were used as experimental strains. The polymerase chain reaction (PCR) and enzyme-linked immunoassay (ELISA) were performed to detect toxin genes and toxin production of different C. difficile ST types, respectively. The count of the colony forming units (CFU) of the strains as conducted by using the brain-heart infusion (BHI) agar plates. The antimicrobial resistance patterns of the strains to eleven kinds of antibiotics were determined by agar dilution method. The antimicrobial resistance genes: gyrA, gyrB and ermB were amplified and sequenced from the stains. Mutations in the resistance genes were analyzed by sequencing. Measure data was compared by Kruskal Wallis Test, differences in the resistance rates in three group were compared using Fisher exact test. Results:ST81 strains were identified as the tcdA-tcdB+/ cdtA-cdtB-toxin type, ST8 and ST42 strains belonged to tcdA+tcdB+/ cdtA-cdtB-toxin type. The toxin production of ST42 strains (41.9) were higher than ST8 (2.4) and ST81 groups (0.83) (all P<0.001). The number of spore quantities of ST81, ST8 and ST42 strains were 494×10 5CFU/ml, 160×10 5CFU/ml and 166×10 5CFU/ml, respectively. The spore quantities of ST81 strains were much higher than that of ST81 and ST42 strains (all P<0.001). From the in vitro susceptibility test, 100% (26/26) ST81 strains were featured as multi-drug resistant (MDR), and they were resistant to moxifloxacin, ceftriaxone, erythromycin and clindamycin. The resistance rates of ST8 strain to moxifloxacin, erythromycin and clindamycin were 9/15, 11/15 and 11/15, respectively. ST81 strains had higher resistance rates to moxifloxacin, clindamycin and erythromycin, compared to ST8 strains ( P=0.001, P=0.005 and P=0.005). All ST42 strains were susceptible to ceftriaxone and 3/14 ST42 strains were resistant to moxifloxacin. ST81 strains had higher resistance rates to ceftriaxone and moxifloxacin than the ST42 strains (both P<0.001). The positive rate of ermB in ST81 strains (100%, 26/26) were higher the ST8 strains (11/15) ( P<0.005). Amino acid mutation analysis showed that ST81and ST8 stains had one amino acid substitution in both GyrA and GyrB, but the amino acid substitutions were different in GyrB between two ST types. ST81 strains had two point-mutations: Thr82 replaced by Ile in GyrA, and Asp426 replaced by Val in GyrB. ST8 strains had point-mutation: Thr82 replaced by Ile in GyrA; Asp426 replaced by Asn in GyrB. For ST42 strains, Thr82 was replaced by Ile in GyrA. Conclusions:ST81 and ST42 strains were MDR. ST81 had higher spore ability, whereas ST42 strains had more virulence. ST81 strains and most of ST8 strains had high level of fluoroquinolones resistance. It is important to supervise persistently these three ST genotypes to prevent further dissemination.

Etiology and prognostic factors of necrotizing enterocolitis in infants after neonatal period with hematochezia / 中国小儿急救医学

Objective:To analyze the etiology and prognostic factors of necrotizing enterocolitis(NEC) in infants after neonatal period with hematochezia.Methods:The clinical data of 62 infants older than 28 days with NEC and hematochezia diagnosed at Beijing Children′s Hospital, Capital Medical University from January 2016 to December 2021 were retrospectively analyzed, summarizing the etiology of NEC in this age group and analyze the factors affecting the prognosis of NEC.According to IgE detection results of food allergens, the infants were divided into milk protein positive group and milk protein negative group.According to the absolute value level of peripheral blood eosinophils, they were divided into increased eosinophils group(≥0.5×10 9/L) and normal eosinophils group(<0.5×10 9/L). They were divided into three groups according to co-infection: NEC group(no co-infection), NEC+ clostridium difficile associated diarrhea(CDAD) group, and NEC+ other infection group(salmonella infection or sepsis). According to different feeding methods, they were divided into normal amino acid group(osmotic pressure 310 mOsm/L), diluted amino acid group(osmotic pressure 233 mOsm/L), and deep hydrolysis group(osmotic pressure 185 mOsm/L). The relief time of clinical symptoms, the recovery time of intestinal gas accumulation, feeding time to achieve physiological requirements, and the length of hospital stay in each group were compared. Results:Among 62 cases, there were 27 males and 35 females.The median age of onset was 1.4(1.2, 2.3) months.The median birth weight was 3.2(2.9, 3.4)kg.Full-term infants accounted for 87.1%.Cesarean accounted for 62.9%.Fifty-three patients(85.5%)had allergic symptoms.Thirteen patients(21.0%)had family history of allergy.Cow milk protein allergy was diagnosed in 29 cases.Thirty-two cases(51.6%) had elevated peripheral blood eosinophils.The hospitalization time of milk protein positive group was longer than that of negative group( P=0.047). The clinical remission rate after hypoallergenic formula feeding for 1 day of increased eosinophils group was higher than that of normal eosinophil group(100.0% vs.65.0%, P=0.002). Ten patients(16.1%)were complicated with clostridium difficile infection, two patients(3.2%) with salmonella enteritis, and four patients(6.5%) with sepsis.Both the hospital stay and feeding time to achieve physiological requirements of NEC+ other infection group were longer than the other two groups( P<0.05). NEC+ CDAD group had a higher rate of repeated hospitalizations(40.0%, P=0.004). The mean recovery time of intestinal gas accumulation was(4.5±2.9)days.After(3.9±3.0)days, hypoallergenic formula feeding started.After one day of feeding, the clinical remission rate was 79.0%.The average time to achieve physiological requirements was(5.8±3.2)days.The clinical symptom relief time of diluted amino acid group was shorter( P=0.006), but there was no statistical difference in feeding time to achieve physiological requirements and hospitalization time between each group( P>0.05). Conclusion:Cow′s milk protein allergy and infection(especially CDAD)are closely related to the occurrence and development of NEC after neonatal period with hematochezia.The administration of diluted amino acid-based formulae close to the osmotic pressure of breast milk and targeted anti-infective therapy could shorten the clinical remission time of NEC and reduce the risk of repeated hospitalization.

Design, synthesis and biological evaluation of 8-hydroxyquinoline derivatives as potential clostridium difficile antibiotics / 药学实践杂志

Objective To find a more effective alternative therapy for antibiotic therapy and fecal microbiota transplantation in current primary treatment of clostridioides difficile infection (CDI) because of the high recurrence rate. Methods A series of 8-hydroxyquinoline derivatives were designed and synthesized based on 8-hydroxyquinoline scarffold. Results The activity test against C. difficile showed that most of the molecules exhibited good antibacterial activity against C. difficile, and compound 6f showed attractive anti-C. difficile activity. Conclusion A new type of 8-hydroxyquinoline derivatives with anti-clostridium difficile was found, which could be used as good lead compounds for further development.

Management of diarrhea after kidney transplantation / 器官移植

Diarrhea is a frequent complication after kidney transplantation, which is a common clinical manifestation of prevalent diseases following multiple types of organ transplantation. The common causes of diarrhea after kidney transplantation include adverse reactions of immunosuppressants, infectious diseases and de novo postoperative inflammatory bowel disease, etc. Diarrhea could seriously affect the quality of life of kidney transplant recipients, and may lead to allograft dysfunction or even death of recipients. Because the causes of diarrhea after kidney transplantation are complicated and probably overlap with each other, along with individual differences among recipients, the etiological diagnosis and targeted treatment of diarrhea after kidney transplantation should follow the principles of gradual and phased treatment. In this article, the epidemiology and harm, common causes and management strategies of diarrhea after kidney transplantation were summarized, aiming to deepen the clinicians' understanding and enhance the diagnosis and treatment levels of diarrhea after kidney transplantation, thereby improving the quality of life and prognosis of kidney transplant recipients.

The fliL gene significantly affects the motility and sporulation abilities of Clostridioides difficile / 生物工程学报

Flagella are the main motility structure of Clostridioides difficile that affects the adhesion, colonization, and virulence of C. difficile in the human gastrointestinal tract. The FliL protein is a single transmembrane protein bound to the flagellar matrix. This study aimed to investigate the effect of the FliL encoding gene flagellar basal body-associated FliL family protein (fliL) on the phenotype of C. difficile. The fliL gene deletion mutant (ΔfliL) and its corresponding complementary strains (: : fliL) were constructed using allele-coupled exchange (ACE) and the standard molecular clone method. The differences in physiological properties such as growth profile, antibiotic sensitivity, pH resistance, motility, and spore production ability between the mutant and wild-type strains (CD630) were investigated. The ΔfliL mutant and the : : fliL complementary strain were successfully constructed. After comparing the phenotypes of strains CD630, ΔfliL, and : : fliL, the results showed that the growth rate and maximum biomass of ΔfliL mutant decreased than that of CD630. The ΔfliL mutant showed increased sensitivity to amoxicillin, ampicillin, and norfloxacin. Its sensitivity to kanamycin and tetracycline antibiotics decreased, and the antibiotic sensitivity partially returned to the level of CD630 strain in the : : fliL strain. Moreover, the motility was significantly reduced in the ΔfliL mutant. Interestingly, the motility of the : : fliL strain significantly increased even when compared to that of the CD630 strain. Furthermore, the pH tolerance of the ΔfliL mutant significantly increased or decreased at pH 5 or 9, respectively. Finally, the sporulation ability of ΔfliL mutant reduced considerably compared to the CD630 strain and recovered in the : : fliL strain. We conclude that the deletion of the fliL gene significantly reduced the swimming motility of C. difficile, suggesting that the fliL gene is essential for the motility of C. difficile. The fliL gene deletion significantly reduced spore production, cell growth rate, tolerance to different antibiotics, acidity, and alkalinity environments of C. difficile. These physiological characteristics are closely related to the survival advantage in the host intestine, which is correlated with its pathogenicity. Thus, we suggested that the function of the fliL gene is closely related to its motility, colonization, environmental tolerance, and spore production ability, which consequently affects the pathogenicity of C. difficile.

Caracterización epidemio-molecular de la infección por Clostridioides difficile en pacientes hospitalizados con cuadro diarreico en instituciones de salud. 2019

La infección por Clostridioides difficile es una de las principales causas de diarrea nosocomial en hospitales del mundo, asociada a antibióticos de amplio espectro. Estudio descriptivo, retrospectivo, de corte transverso, de tipo censal. Se estudiaron 281 muestras de pacientes hospitalizados con la infección, se analizaron características socio-demográficas, clínicas y se caracterizó molecularmente al patógeno. Como resultado, se obtuvieron pacientes con la infección con una mediana de edad de 64 años siendo el 61,5 % del sexo masculino. El promedio de presentación del cuadro diarreico fue de 5 días, con tratamiento antimicrobiano de 8 días e internación de 15 días. El 94% tuvo tratamiento antimicrobiano previo, y el 6% estuvo expuesto a algún factor de riesgo. Los antimicrobianos más utilizados solos o en combinación fueron betalactámicos, fluoroquinolonas, vancomicina y carbapenémicos. Las áreas de internación con mayor frecuencia de presentación de la infección fueron las unidades de Clínica Médica, Traumatología, Geriatría y Unidad de Terapia Intensiva. La prevalencia de C. difficile toxigénico fue de 14%, y de esta frecuencia el 100% presentó toxinas TcdA y TcdB, con ausencia de toxinas binarias y deleción del gen tcdC. Se constató la presencia grupos clonales en la misma unidad de internación y misma institución de salud. El 100% de las cepas resultaron susceptibles a los antibióticos de elección para la infección. La prevalencia de la infección y presencia de perfiles clonales detectadas revelan la necesidad de un mejoramiento en el sistema de control de infecciones, así como del fortalecimiento y vigilancia de la resistencia antimicrobiana.

Clostridioides difficile infection is one of the main causes of nosocomial diarrhea in hospitals worldwide, associated with broad-spectrum antibiotics. Descriptive, retrospective, cross-sectional, census-type study. Two hundred eighty-one samples from patients hospitalized with the infection were studied, sociodemographic and clinical characteristics were analyzed and the pathogen was characterized molecularly. As a result, patients with the infection had a median age of 64 years and 61.5% male. The average presentation of diarrhea was 5 days, antimicrobial treatment 8 days and hospitalization 15 days. Ninety four percent had previous antimicrobial treatment, and 6% were exposed to some risk factor. The most commonly used antimicrobials alone or in combination were beta-lactams, fluoroquinolones, vancomycin, and carbapenems. The hospitalization areas with the highest frequency of presentation of the infection were the Clinical Medicine, Traumatology, Geriatrics and Intensive Care Unit units. The prevalence of toxigenic C. difficile was 14%, and of this frequency, 100% presented TcdA and TcdB toxins, with the absence of binary toxins and deletion of the tcdC gene. The presence of clonal groups was verified in the same hospitalization unit and the same health institution. All the strains were susceptible to the antibiotics of choice for the infection. The prevalence of infection and the presence of clonal profiles detected reveal the need for improvement of the infection control system, as well as of the strengthening and surveillance of antimicrobial resistance.

A case-control study of Clostridioides difficile symptomatic infections in a pediatric cancer hospital / Um estudo caso-controle de infecções sintomáticas por Clostridioides difficile em um hospital oncológico pediátrico

Abstract Objective: The aim of this study was to analyze and identify documented infections and possible risk factors for Clostridioides difficile infections in children with cancer. Methods: This is a retrospective case-control study, carried out in a pediatric cancer hospital, covering the years 2016-2019. Matching was performed by age and underlying disease, and for each case, the number of controls varied from 1 to 3. Logistic regression models were used to assess risk factors. Results: We analyzed 63 cases of documented infection by C. difficile and 125 controls. Diarrhea was present in all cases, accompanied by fever higher than 38°C in 52.4% of the patients. Mortality was similar among cases (n=4; 6.3%) and controls (n=6; 4.8%; p=0.7). In all, 71% of patients in the case group and 53% in the control group received broad-spectrum antibiotics prior to the infection. For previous use of vancomycin, the Odds Ratio for C. difficile infection was 5.4 (95% confidence interval [95%CI] 2.3-12.5); for meropenem, 4.41 (95%CI 2.1-9.2); and for cefepime, 2.6 (95%CI 1.3-5.1). For the antineoplastic agents, the Odds Ratio for carboplatin was 2.7 (95%CI 1.2-6.2), melphalan 9.04 (95%CI 1.9-42.3), busulfan 16.7 (95%CI 2.1-134.9), and asparaginase 8.97 (95%CI 1.9-42.9). Conclusions: C. difficile symptomatic infection in children with cancer was associated with previous hospitalization and the use of common antibiotics in cancer patients, such as vancomycin, meropenem, and cefepime, in the last 3 months. Chemotherapy drugs, such as carboplatin, melphalan, busulfan, and asparaginase, were also risk factors.

RESUMO Objetivo: Analisar e identificar infecções documentadas e possíveis fatores de risco para infecções por Clostridioides difficile em crianças com câncer. Métodos: Estudo retrospectivo caso-controle em um hospital pediátrico oncológico, que abrangeu os anos de 2016-2019. O pareamento foi realizado por idade e doença de base e, para cada caso, o número de controles variou de um a três. Modelos de regressão logística foram utilizados para avaliar os fatores de risco. Resultados: Analisamos 63 casos de infecção documentados por C. difficile e 125 controles. A diarreia esteve presente em todos os casos, acompanhada de febre acima de 38°C em 52,4% dos pacientes. A mortalidade foi semelhante entre casos (n=4, 6,3%) e controles (n=6, 4,8%; p=0,7). No grupo caso, 71% dos pacientes e, no grupo controle, 53% deles receberam antibióticos de amplo espectro antes da infecção. Para uso prévio de vancomicina, a Odds Ratio para infecção por C. difficile foi de 5,4 (intervalo de confiança [IC95%] 2,3-12,5); para meropenem, 4,41 (IC95% 2,1-9,2) e, para cefepima, 2,6 (IC95% 1,3-5,1). Para os agentes antineoplásicos, a razão de chances para carboplatina foi de 2,7 (IC95% 1,2-6,2), para melfalano de 9,04 (IC95% 1,9-42,3), para bussulfano de 16,7 (IC95% 2,1-134,9) e, para asparaginase, de 8,97 (IC95% 1,9-42,9). Conclusões: A infecção sintomática por C. difficile em crianças com câncer associou-se à internação prévia e ao uso de antibióticos como vancomicina, meropenem e cefepime nos últimos três meses. Os quimioterápicos carboplatina, melfalano, bussulfano e asparaginase também foram fatores de risco.

Aetiology of hospital-acquired diarrhoea in under-five children from an urban hospital in East Delhi, India

Background & objectives: Majority of the studies of hospital-acquired diarrhoea conducted in Western countries have focused on the detection of Clostridium difficile in stool samples. Limited Asian and Indian literature is available on hospital-acquired diarrhoea. This study was aimed to describe the aetiological profile for hospital-acquired diarrhoea in children aged below five years. Methods: One hundred children aged one month to five years who developed diarrhoea (?3 loose stools for >12 h) after hospitalization for at least 72 h were enrolled. Children who were prescribed purgatives or undergoing procedures such as enema and endoscopy or those with underlying chronic gastrointestinal disorders such as celiac disease and inflammatory bowel disease were excluded from the study. Stool samples from the enrolled children were subjected to routine microscopic examination, modified Ziel- Nielson (ZN) staining for Cryptosporidium and culture for various enteropathogens. Multiplex PCR was used to identify the strains of diarrhoeagenic Escherichia coli. Rotavirus detection was done using rapid antigen kit. Toxins (A and B) of C. difficile were detected using enzyme immunoassay. Results: Of the 100 samples of hospital-acquired diarrhoea analysed, diarrhoeagenic E. coli (DEC) was found to be the most common organism, detected in 37 per cent of cases (enteropathogenic E. coli-18%, enterotoxigenic E. coli-8%, enteroaggregative E. coli-4% and mixed infections-7%). Cryptosporidium was detected in 10 per cent of cases. Rotavirus was detected in six per cent and C. difficile in four per cent of cases. Interpretation & conclusions: The findings of this study suggest that the aetiological profile of hospital- acquired diarrhoea appears to be similar to that of community-acquired diarrhoea, with DEC and Cryptosporidium being the most common causes. The efforts for the prevention and management of hospital-acquired diarrhoea should, thus, be directed towards these organisms.

Trasplante de microbiota fecal por colonoscopía en paciente mayor de 65 años con infección recurrente de Clostridioides difficile: aún una estrategia subutilizada / Fecal microbiota transplantation in an older patient with Clostridioides difficile recurrent infection. Report of one case

Clostridioides difficile infection (CDI) is a major public health problem and responsible for significant morbidity and mortality. Eighty percent of CDIs occur in adults older than 65 years of age due to a decreased gastrointestinal microbial diversity, immunosenescence and frailty. Thus, the most reported risk factor for recurrent CDI is older age since nearly 60% of cases occur in individuals aged ≥ 65 years. Fecal microbiota transplantation (FMT) is a highly cost-effective alternative to antibiotic treatment for patients with recurrent CDI. We report a 75-year-old male with recurrent CDI, who received a FMT after several unsuccessful antimicrobial treatments. He had a satisfactory evolution after the procedure and remained without diarrhea during the ensuing five months.

Infecciones por Clostridioides (Clostridium) difficile en un hospital geriátrico de Costa Rica / Clostridioides (Clostridium) difficile infections in a geriatric hospital in Costa Rica

Objetivo: Describir las características de los pacientes adultos mayores con diagnóstico de infecciones por Clostridium difficile en un hospital geriátrico en Costa Rica con el propósito de contribuir a mejorar su manejo y llevar a una reducción de la morbimortalidad y costos asociados a su atención. Métodos: Se realizó un estudio observacional retrospectivo con información demográfica y clínica de 141 pacientes admitidos en el Hospital Nacional de Geriatría y Gerontología Dr. Raúl Blanco Cervantes de Costa Rica del 2015 al 2018, quienes presentaron una prueba inmunocromatográfica de detección de antígeno y/o toxinas de C. difficile positiva en heces diarreicas. Las variables continuas se compararon mediante una prueba de ANOVA, mientras que las categóricas, por una prueba exacta de Fisher. Los factores de riesgo para cada uno de los grupos se evaluaron por análisis univariante. Los valores de p < 0,05 se consideraron estadísticamente significativos con un 95% de confianza. Resultados: Se estudiaron 141 pacientes con diarrea asociada a C. difficile. Los pacientes tenían una edad promedio de 83 años y 57% eran mujeres. Un 35% de los casos eran de origen comunitario y 27% fueron severos. El consumo de antimicrobianos fue dado principalmente por cefalosporinas y fluoroquinolonas. El tratamiento más utilizado fue el metronidazol (81%) y la mortalidad relacionada con la infección por C. difficile a los 30 días fue de un 35%. Conclusiones: Este es el primer reporte epidemiológico de infección por C. difficile que describe a un grupo de pacientes geriátricos hospitalizados y sus factores de riesgo asociados, que pone en manifiesto un porcentaje importante de casos comunitarios y graves, lo que llama a establecer guías locales y grupos específicos para el tratamiento y prevención de dicha infección.

Aim: To describe the characteristics of elder patients diagnosed with Clostridioides (Clostridium) difficile infection in a geriatric hospital in Costa Rica. Methods: A retrospective observational study was done with demographic and clinical information from 141 patients admitted in the National Geriatric and Gerontology Hospital of Costa Rica from 2015 to 2018, who presented a positive immunochromatographic test for the detection of C. difficile antigen and/or toxins in diarrheic feces. Continuous variables were compared through one-way ANOVA test, while categorical variables were compared using Fisher's exact test. The risk factors for each of the groups were evaluated by univariate analysis. P values < 0.05 were considered statistically significant with 95% confidence. Results: We studied 141 patients with diarrhea associated with C. difficile, the average age of the patients was 83 years and 75% were women. 35% of the cases were community acquired and 27% were severe cases. Antimicrobial consumption was given mainly by cephalosporins and fluoroquinolones. Metronidazol was the most used treatment (81%) and the C. difficile associated mortality 30 days post infection was 35%. Conclusion: This is the first epidemiological report of C. difficile infection in elderly hospitalized population. Also, it evidences an important percentage of community acquired and severe cases, calling for the establishment of local treatment and prevention guidelines for this infection.

Diagnóstico microbiológico de infección por Clostridioides difficile en épocas de COVID-19 en un hospital pediátrico de alta complejidad / Microbiological diagnosis of Clostridioides difficile infection in times of COVID-19 in a high complexity pediatric hospital

Resumen La infección por Clostridioides difficile (ICD) puede variar desde diarrea hasta megacolon tóxico. Los objetivos del trabajo fueron mostrar la variación en el número de casos diagnosticados de ICD en este laboratorio entre 2020, cuando comenzó la pandemia de COVID-19 y 2019 y 2021 y detallar los casos precedidos por la infección de SARS-CoV-2. El presente es un estudio retrospectivo observacional en el que se registraron el número total de muestras procesadas con sospecha de ICD y el de positivas y los antecedentes clínicos de pacientes con ICD hasta dos meses después de su diagnóstico de COVID-19. Durante 2020 se procesaron menos muestras que en 2019 y 2021; sin embargo, el porcentaje de positividad fue de 13,1%, 7,2% y 7,8%, respectivamente. Esto pudo deberse a mejoras en el criterio clínico al momento de seleccionar las muestras con sospecha de ICD.

Abstract Clostridioides difficile infection (CDI) can cause anything from diarrhea to toxic megacolon. The objectives of this study were: to show the variation in the number of diagnosed cases of CDI in this center, comparing 2020, when the COVID-19 pandemic began, with 2019 and 2021 and to detail cases preceded by SARS-CoV-2 infection. This is an observational retrospective study in which the total number of samples processed with suspected CDI were recorded. The positive ones and the clinical history of patients with a diagnosis of CDI up to two months after their diagnosis of SARS-CoV-2 infection were recorded as well. During 2020 a smaller number of samples were processed. However, during this year the percentage of positivity was 13.1% vs. 7,2% and 7.8% during 2019 and 2021, respectively. It is believed that this may have been due to improvements in clinical suspicion and sample selection for CDI diagnosis.

Resumo A infecção por Clostridioides difficile (ICD) pode causar desde diarreia até megacólon tóxico. Os objetivos desta apresentação foram: mostrar a variação do número de casos diagnosticados de ICD neste laboratório, entre 2020 quando começou a pandemia de COVID-19 e 2019 e 2021 e, detalhar os casos precedidos pela infecção por SARS-CoV-2. Esse estudo foi retrospectivo observacional e foram registrados: o número total de amostras processadas com suspeita de ICD e de amostras positivas e os antecedentes clínicos daqueles pacientes com diagnóstico de ICD até dois meses após o diagnóstico de COVID 19. Durante 2020, foram processadas menos amostras do que em 2019 e 2021; no entanto, o percentual de positividade foi de 13,1%, 7,2% e 7,8%, respectivamente. Isso pode ter sido resultado de melhorias no critério clínico na hora de selecionar as amostras com suspeita de ICD.

Clostridium Difficile is associated with High Mortality among Cirrhotics in India

Background: To assess the prevalence and impact of Clostridium difficile infection (CDI) in hospitalized patients with cirrhosis in India. Methods: In this prospective observational study from June 2015 to March 2016, all hospitalized patients with cirrhosis and acute diarrhea at the time of admission or during hospitalization were included. We studied hospitalized patients with cirrhosis without diarrhea during the same period to detect asymptomatic colonizers.Stool samples were tested for CDI, bacterial cultures, and parasite microscopy in patients with diarrhea.CDI was detected using a stool PCR test that detects the pathogenicity locus of toxigenic Clostridium difficile gene. We analysed the impact of CDI on hospital outcomes and also assessed the risk factors for acquiring CDI. Result: Among 92 hospitalized cirrhotic patients with acute diarrhea [male: 74; median age: 50 (range 19 to 80) years; Child’s class A: B: C: 8:41:43; median MELD score: 18 (range 6 to 44)], 6 (6.5%) had CDI by positive stool PCR. Use of antibiotics (100% CDI Vs 55.8% non-CDI, p= 0.04) and steroids (50% CDI vs 10.5% non-CDI, p =0.028) emerged as risk factors for CDI among cirrhosis patients. Two of the 6 patients (33.3%) with CDI as compared to 6/86 patients (7%) with no CDI died (p-value: 0.08).There were no asymptomatic colonizers amongst 35 hospitalized cirrhosis patients without diarrhea.Conclusions: C. difficile, although uncommon, was an important cause of mortality in cirrhosis patients hospitalized with diarrhea in India.Prior use of antibiotics or steroids were identified as risk factors for CDI.

Umbral del ciclo de amplificación del gen de la toxina B de Clostridioides difficile como predictor de gravedad / Clostridioides difficile toxin B gen amplification cycle threshold as a predictor of severity

INTRODUCCIÓN: La infección por Clostridioides dfficile (ICD) es la principal causa de diarrea nosocomial, generalmente asociada al consumo de antimicrobianos. Esta infección puede causar desde diarrea no complicada hasta colitis pseudomembranosa o megacolon tóxico. Estudios recientes han intentado relacionar el valor el ciclo umbral (Ct) de la RT-PCR con la mortalidad, como un método rápido, sencillo, objetivo y eficaz. OBJETIVO: Evaluar el Ct como predictor de mala evolución en pacientes con y sin criterio clínico de dicha gravedad. PACIENTES Y MÉTODOS: Realizamos un estudio retrospectivo entre enero 2015 y diciembre 2018, incluyendo todos los pacientes del área de referencia del Hospital Universitario de Canarias en Tenerife (396.483 habitantes) en pacientes con criterios clínicos de gravedad (de acuerdo a la Guía para la Práctica Clínica de la enfermedad por C. dfficile de la Sociedad de Epidemiología del Cuidado de la Salud de América (SHEA) y la Sociedad de Enfermedades Infecciosas de Norteamérica (IDSA) y pacientes sin criterios clínicos de gravedad evaluando el Ct como predictor de mala evolución. RESULTADOS: Se diagnosticó un total de 202 episodios de ICD. El 77,7% (n = 157) presentó criterios clínicos de gravedad. La presencia de colitis ulcerosa (p < 0,001), fiebre (p < 0,001), leucocitosis (p < 0,001), neutrofilia (p < 0,001), creatininemia (p = 0,005) se presentaron como factores de riesgo para el desarrollo de ICD grave. El sexo femenino, la institucionalización, el ingreso previo y el exitus se describieron con mayor frecuencia en el grupo con ICD-G, no encontrando diferencias significativas. No encontramos diferencias respecto a los días de estancia previa, o de estancia post-ICD, aunque en este último, la media fue mayor en el caso de los pacientes con ICD-G. No se encontraron diferencias significativas en cuanto al Ct en ambos grupos; siendo sólo un punto menor en pacientes con criterio de gravedad (Ct = 26,1) que sin criterios de gravedad (Ct = 27,4) (p = 0,326).

BACKGROUND: Clostridioides dfficile infection (CDI) is the main cause of nosocomial diarrhea, generally associated with the use of antibiotics. This infection can cause uncomplicated diarrhea to pseudomembranous colitis or toxic megacolon. Recent studies have attempted to relate the threshold cycle (Ct) value of RT-PCR with mortality, as a fast, simple, objective and efficient method. AIM: To evaluate Ct as a predictor of poor outcome in patients with C. dfficile disease with/without clinical signs of severity. METHODS: We carried out a retrospective study between January 2015 and December 2018, including all patients in the reference area of the Hospital Universitario de Canarias in Tenerife (396,483 inhabitants) in patients with clinical criteria of severity and patients without clinical severity criteria (according to the guide for the clinical practice of CDI of the Society of Healthcare Epidemiology of America (SHEA) and the Infectious Diseases Society of North America (IDSA). RESULTS: A total of 202 CDI episodes were diagnosed. 77.7% (n = 157) presented clinical severity criteria. The presence of ulcerative colitis (p < 0.001), fever (p < 0.001), leukocytosis (p < 0.001), neutrophilia (p < 0.001), creatininemia (p = 0.005) were presented as risk factors for the development of severe CDI (S-CDI). Female sex, institutionalization, previous admission and death were described more frequently in the group with S-CDI, not finding significant differences. We found no differences with respect to the days of previous stay, or of post-CDI stay, although in the latter, the mean was higher in the case of S-CDI patients. No significant differences were found in terms of Ct in both groups; being only one point lower in patients with severity criteria (Ct = 26.1) than without severity criteria (Ct = 27.4) (p = 0.326). CONCLUSION: Based on the results of our study, it has not been possible to systematically implement the Ct value as a predictor of severity to the clinical report, and it is not possible to extrapolate this predictive variable from S-CDI and standardize the Ct value as a predictor of severity. Conclusion: Basándonos en los resultados de nuestro estudio, no ha sido posible la implementación sistemática del valor Ct como predictor de gravedad al informe clínico, no siendo posible extrapolar esta variable predictora de enfermedad por C difficile-G y estandarizar el valor Ct como factor predictor de gravedad.