scholarly journals Diarrhoea caused by Clostridium difficile in patients with postoperative subhepatic abscess

2008 ◽  
Vol 65 (3) ◽  
pp. 249-254 ◽  
Author(s):  
Predrag Stojanovic ◽  
Branislava Kocic
Keyword(s):  
Clostridium Difficile ◽  
Wound Infection ◽  
Day Treatment ◽  
Toxin B ◽  
Causal Therapy ◽  
Patient Reported ◽  
Stool Samples ◽  
Toxigenic Strains ◽  
Subhepatic Abscess ◽  
Operative Wound

Background. Toxigenic strains of Clostridium difficile in the majority of cases cause disease of the intestinal tract of hospitalized patients. For a long time, Clostridium difficile was considered to produce both types of toxins (A+/B+ strain), however, the investigations conducted in the last ten years point to the existence of clinically significant isolates which produce only toxin B, i.e. toxin A negative / toxin B positive (A-/B+ strain) Clostridium difficile. Case report. We presented the case of a patient admitted to the Surgery Clinic, Clinical Center Nis due to the presence of calculus in the ductus choledochus. Twenty-four hours after the surgical intervention for calculus removal, the first signs of the operative wound infection began to appear. In the course of infection treatment, different antibiotics were administered (cefuroxine, ciprofloxacin, vancomycin, imipenem). After making etiological microbiological diagnosis and application of antibiotics according to antibiogram results, the signs of the operative wound infection began to withdraw, but the patient reported the abdominal pain and liquid stools with traces of blood (up to 17 stools per day). By microbiological examination, Clostridium difficile was cultivated and the presence of toxin B was detected in the stool samples. The patient was sent to the Clinic for Infectious Diseases, where the causal therapy of mitronidazol was administered. Liquid and electrolytes were made up by substitution therapy. After the eight-day-treatment, the patient felt much better, and diarrheas stopped on the 10th day of the therapy application. Conclusion. Our results have shown that toxingen strains Clostridium difficile are present in our country so this bacterium sort have to be considered in differential causal diagnosis of diarrhoea syndrom. Considering that it can cause difficult form of the disease, it is an obligation to establish the presence of some toxins of Clostridium difficile in stool samples of patients and/or production of some toxins in liquid culturate of isolates to provide data for the presence of strains which produce only toxin B.

scholarly journals Detection of enterotoxin A and cytotoxin B, and isolation of Clostridium difficile in piglets in Minas Gerais, Brazil

2011 ◽  
Vol 41 (8) ◽  
pp. 1430-1435 ◽  
Author(s):  
Rodrigo Otávio Silveira Silva ◽  
Felipe Masiero Salvarani ◽  
Eduardo Coulaud da Costa Cruz Júnior ◽  
Prhiscylla Sadanã Pires ◽  
Renata Lara Resende Santos ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Binary Toxin ◽  
Toxin Gene ◽  
Toxin A ◽  
Toxin B ◽  
Fecal Samples ◽  
Newborn Piglets ◽  
Pig Farms ◽  
Stool Samples ◽  
Toxigenic Strains

Clostridium difficile has emerged as a major cause of neonatal colitis in piglets, displacing classic bacterial pathogens. However, there is no information regarding the distribution of this microorganism in pig farms in Brazil. In the present study, the presence of toxins A/B and of C. difficile strains in stool samples from 60 diarrheic or non-diarrheic newborn piglets (one to seven days old), from 15 different farms, was studied. The presence of toxins A/B was detected by ELISA and PCR was used to identify toxin A, toxin B and binary toxin gene in each isolated strain. C. difficile A/B toxins were detected in ten samples (16.7%). Of these, seven were from diarrheic and three were from non-diarrheic piglets. C. difficile was recovered from 12 out of 60 (20%) fecal samples. Of those, three strains were non-toxigenic (A-B-) and nine were toxigenic. Of the nine toxigenic strains, four were A+B+ strains and five were A-B+ strains. The presence of binary toxin observed in the present study was much higher (50%) than in previously reported studies. All three non-toxigenic strains were isolated from otherwise healthy piglets. The results suggest the occurrence of neonatal diarrhea by C. difficile in farms in Brazil.

scholarly journals Identification of Toxin A-Negative, Toxin B-Positive Clostridium difficile by PCR

1998 ◽  
Vol 36 (8) ◽  
pp. 2178-2182 ◽  
Author(s):  
Haru Kato ◽  
Naoki Kato ◽  
Kunitomo Watanabe ◽  
Naoichi Iwai ◽  
Haruhi Nakamura ◽  
...  
Keyword(s):  
Cell Culture ◽  
Clostridium Difficile ◽  
Pcr Amplification ◽  
Enzyme Linked Immunosorbent Assay ◽  
Pcr Assay ◽  
Toxin A ◽  
Toxin B ◽  
Cell Monolayers ◽  
Toxigenic Strains ◽  
Cell Culture Assay

Toxigenic strains of Clostridium difficile have been reported to produce both toxins A and B nearly always, and nontoxigenic strains have been reported to produce neither of these toxins. Recent studies indicate that it is not always true. We established a PCR assay to differentiate toxin A-negative, toxin B-positive (toxin A−, toxin B+) strains from both toxin-positive (toxin A+, toxin B+) strains and both toxin-negative (toxin A−, toxin B−) strains as an alternative to cell culture assay and enzyme-linked immunosorbent assay (ELISA). By using the PCR primer set NK11 and NK9 derived from the repeating sequences of the toxin A gene, a shorter segment (ca. 700 bp) was amplified from toxin A−, toxin B+ strains compared to the size of the segment amplified from toxin A+, toxin B+ strains (ca. 1,200 bp), and no product was amplified from toxin A−, toxin B− strains. We examined a total of 421 C. difficile isolates by PCR. Of these, 48 strains showed a shorter segment by the PCR, were negative by ELISAs for the detection of toxin A, and were positive by cell culture assay. Although the cytotoxin produced by the toxin A−, toxin B+ strains was neutralized by anti-toxin B serum, the appearance of the cytotoxic effects on Vero cell monolayers was distinguishable from that of toxin A+, toxin B+ strains. By immunoblotting, the 44 toxin A−, toxin B+ strains were typed to serogroup F and the remaining four strains were serogroup X. Pulsed-field gel electrophoresis separated the 48 strains into 19 types. The PCR assay for the detection of the repeating sequences combined with PCR amplification of the nonrepeating sequences of either the toxin A or the toxin B gene is indicated to be useful for differentiating toxin A−, toxin B+ strains from toxin A+, toxin B+ and toxin A−, toxin B− strains and will contribute to elucidation of the precise role of toxin A−, toxin B+ strains in intestinal diseases.

scholarly journals A Rapid, Accurate, Single Molecule Counting Method Detects Clostridium difficile Toxin B in Stool Samples

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Sadanand Gite ◽  
Destiny Archambault ◽  
Michael P. Cappillino ◽  
David Cunha ◽  
Victoria Dorich ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Single Molecule ◽  
Counting Method ◽  
Toxin B ◽  
Clostridium Difficile Toxin ◽  
Stool Samples ◽  
Clostridium Difficile Toxin B

scholarly journals Performing Reflexive Toxin A/B Enzyme Immunoassay in a Two-Step Algorithm for the Diagnosis of Clostridium difficile Infection Has Limited Clinical Utility

2018 ◽  
Author(s):  
M Mounajjed ◽  
R Pease ◽  
B Jung-Hynes ◽  
N Safdar ◽  
D Chen
Keyword(s):  
Clostridium Difficile ◽  
Enzyme Immunoassay ◽  
Electronic Medical Records ◽  
Medical Records ◽  
Clinical Utility ◽  
Nucleic Acid Amplification ◽  
Toxin B ◽  
Clinical Sensitivity ◽  
Stool Samples ◽  
Step Algorithm

ABSTRACTThe differentiation of Clostridium difficile infection (CDI) from colonization is challenged by the suboptimal clinical specificity of nucleic acid amplification tests (NAAT). In this study, we examined the utility of testing for toxin via enzyme immunoassay (EIA) in specimens already tested by NAAT for the diagnosis of CDI in an attempt to differentiate colonization from infection. We tested 59 stool samples for the presence of C. difficile toxin B gene by NAAT followed by EIAs for glutamate dehydrogenase (GDH EIA) and toxins A and B (Toxin EIA). Two infectious disease physicians independently reviewed the patients’ electronic medical records retrospectively to categorize each patient as CDI-Likely, CDI-Unlikely, or CDI-Indeterminate. Clinical sensitivities and specificities were calculated using 3 definitions of “true” CDI status, being: (1) concordance between both reviewers, (2) concordance, and CDI-Indeterminate/discordant cases classified as CDI-Likely, and (3) concordance, and CDI-Indeterminate/discordant cases classified as CDI-Unlikely. Based on these definitions, clinical sensitivity and specificity for NAAT was 100% and 49-94%, GDH EIA was 83-85% and 43-89%, and Toxin EIA was 39-42% and 83-100%, respectively. 85% (22 of 26) of patients who were NAAT-positive but Toxin EIA-negative symptomatically benefited from treatment for CDI. The addition of EIA to NAAT for CDI diagnosis had limited utility for differentiating colonization from CDI and could have led to under treatment of patients with CDI.

scholarly journals Distribution of Clostridium Difficile Ribotypes in Macedonian Patients and their Antimicrobial Susceptibility

2019 ◽  
Vol 7 (12) ◽  
pp. 1896-1899
Author(s):  
Kiril Mihajlov ◽  
Aneta Andreska ◽  
Nadica Ristovska ◽  
Tatjana Grdanoska ◽  
Elena Trajkovska-Dokic
Keyword(s):  
Clostridium Difficile ◽  
Antibiotic Use ◽  
Nosocomial Pathogen ◽  
Toxin B ◽  
Columbia Blood Agar ◽  
Hospitalised Patients ◽  
Pcr Ribotyping ◽  
Stool Samples ◽  
Vitek 2 ◽  
Final Confirmation

BACKGROUND: Clostridium difficile is a major nosocomial pathogen. In Europe, this bacterium is mostly characterised by PCR ribotyping. Most of the Clostridium difficile infections (CDI) are treated with vancomycin or metronidazole, although prolonged antibiotic use is considered as one of the main risk factors for CDI.AIM: This study aimed to detect the presence of various C. difficile ribotypes in hospitalised patients and to investigate their toxigenicity and antibiotic susceptibility.MATERIAL AND METHODS: All stool samples obtained from each patient were inoculated on Columbia blood agar and cycloserine cefoxitine fructose agar (CCFA) for isolation of C. difficile. Glutamate dehydrogenase and toxins A and B were investigated by immunochromatographic tests. Final confirmation of the isolates was performed by Vitek 2 and MALDI-TOF. A total of 21 isolates were collected for further investigation. PCR ribotyping was performed as described by Janezic and Rupnik. PCR ribotype profiles were analysed using software (Bionumerics, Applied Maths). Antibiotic susceptibility was determined by E-tests for metronidazole, vancomycin, tetracycline, clindamycin, erythromycin, imipenem, ciprofloxacin and moxifloxacin.RESULTS: About 48% of C. difficile isolates belonged to ribotype 001/072. So, this ribotype was the most common ribotype in this study. The remaining 52% of C. difficile isolates consisted of 10 different ribotypes: 017, SLO 160, SLO 187, SLO 120, 255/258, 014/020, 046, 002, 070 and 027. Furthermore, 20 (95.2 %) out of 21 isolates of C. difficile were toxigenic. Toxins A and B were detected simultaneously in 90.5 % of C. difficile isolates. Two isolates from the ribotype 017 were toxin B positive only. Treatments with any of the following antimicrobials: clindamycin, erythromycin, ciprofloxacin and moxifloxacin (as well as many other antibiotics), could be a risk factor for CDI due to the high resistance of the strains in this study. About 90% of the strains from the most common ribotype 001/072 have MICs for clindamycin and erythromycin > 256 µg/ml. CONCLUSION: All strains isolated are highly resistant to ciprofloxacin. All strains were susceptible to vancomycin (median MIC was 0.63 µg/ml) and metronidazole (median MIC was 0.084 µg/ml), so these two antimicrobials remain optimal treatment option for CDI.

scholarly journals Clostridium difficile Toxin B Induces Apoptosis in Intestinal Cultured Cells

1998 ◽  
Vol 66 (6) ◽  
pp. 2660-2665 ◽  
Author(s):  
Carla Fiorentini ◽  
Alessia Fabbri ◽  
Loredana Falzano ◽  
Andrea Fattorossi ◽  
Paola Matarrese ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Cultured Cells ◽  
Chromatin Condensation ◽  
Significant Proportion ◽  
Intestinal Cells ◽  
Single Chain ◽  
Toxin B ◽  
Clostridium Difficile Toxin ◽  
Toxigenic Strains ◽  
Clostridium Difficile Toxin B

ABSTRACT Toxigenic strains of the anaerobic bacterium Clostridium difficile produce at least two large, single-chain protein exotoxins involved in the pathogenesis of antibiotic-associated diarrhea and colitis. Toxin A (CdA) is a cytotoxic enterotoxin, while toxin B (CdB) is a more potent cytotoxin lacking enterotoxic activity. This study dealt with CdB, providing the first evidence that intestinal cells exposed to this toxin exhibit typical features of apoptosis in that a significant proportion of the treated cells displayed nuclear fragmentation and chromatin condensation. In keeping with ultrastructural data, CdB-treated cells showed the typical flow cytometric hallmark of apoptosis consisting of a distinct sub-G1 peak. The CdB-induced apoptotic response was dose and time dependent and not simply due to the actin-disrupting effect of the toxin or to the subsequent impairment of cell anchorage. Rather, the inhibition of proteins belonging to the Rho family due to CdB seems to play a role in the induction of apoptosis in intestinal cells. The origin of cells and the growth rate may also be cofactors relevant to such a response.

scholarly journals Genes Encoding Toxin ofClostridium difficilein Children with and without Diarrhea

Scientifica ◽  
2014 ◽  
Vol 2014 ◽  
pp. 1-4 ◽  
Author(s):  
Victor R. C. Merino ◽  
Viviane Nakano ◽  
Sydney M. Finegold ◽  
Mario J. Avila-Campos
Keyword(s):  
16S Rrna ◽  
Binary Toxin ◽  
Genetic Profile ◽  
Rrna Gene ◽  
Toxin B ◽  
Genes Encoding ◽  
Gene 16S Rrna ◽  
Stool Samples ◽  
Toxigenic Strains ◽  
The 16S Rrna Gene

The presence of gene 16S rRNA and genes encoding toxin A (tcdA), toxin B (tcdB), and binary toxin (cdtA/cdtB) ofClostridium difficilein stool samples from children with (110) and without (150) diarrhea was determined by using a TaqMan system. Fifty-seven (21.9%) out of 260 stool samples harbored the 16S rRNA gene. The genetic profile oftcdA+/tcdB−andcdtA+/cdtB+was verified in oneC. difficile-positive diarrhea sample and oftcdA+/tcdB+in threeC. difficile-positive nondiarrhea samples. The presence oftcdA+/tcdB+in stools obtained from children without diarrhea, suggests that they were asymptomatic carriers of toxigenic strains.

Rapid detection of toxigenic Clostridium difficile from stool samples by a nested PCR of toxin B gene

1999 ◽  
Vol 41 (2) ◽  
pp. 145-149 ◽  
Author(s):  
R. Alonso ◽  
C. Muñoz ◽  
S. Gros ◽  
D. García de Viedma ◽  
T. Peláez ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Nested Pcr ◽  
Rapid Detection ◽  
Toxin B ◽  
Stool Samples
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