Isolation and molecular characterization of Clostridium difficile strains from patients and the hospital environment in Belarus (original) (raw)
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Journal of Medical Microbiology, 2015
Clostridium difficile infection (CDI) leads to considerable morbidity and mortality among hospitalized patients. Faecal specimens from 1110 hospitalized patients suspected for CDI were cultured for isolation of C. difficile and characterization of virulence genes. PCR was carried out for toxigenic genes tcdA, tcdB, cdtA and cdtB and PCR-RFLP for fliC and slpA genes. Of 174 (15.7 %) C. difficile isolates, 121 (69.5 %) were toxigenic, amongst which 68 (56.2 %) also had both tcdA and tcdB genes. The remaining 53 (43.8 %) of the isolates also had at least one of the toxin genes. Binary toxin genes (cdtA and cdtB) with only one of the two components were present in 16 (9.2 %) of the 174 isolates. The other virulence genes-fliC and slpA-were present in 100 % of the isolates. The most frequent PCR-RFLP type of fliC gene was type I (n5101), followed by type VII (n549) and type III (n524). The slpA gene presented with three combinations of patterns. Characterization of virulence genes in C. difficile isolates is of extreme importance for epidemiological surveillance and control of outbreaks owing to the capacity of this bacterium to adapt to new environmental circumstances, leading to the emergence of new epidemic strains.
African Journal of Microbiology Research, 2013
Detection of the source of Clostridium difficile for the control of nosocomial infections produced by these bacteria is very important. For this reason 84 C. difficile isolated from 250 stool samples of symptomatic patients, staff, asymptomatic patients at first day of admission, the same patients after seven days of hospitalization and 135 samples from their hospital environment were typed by AP-PCR. In addition to conventional standard tests, gas liquid chromatography was also used as complementary test to identify C. difficile isolates. All isolates were separated into 12 genotypes, with 31% falling into group I. Genotypes VIII, X and XII were found only in isolates of symptomatic patients while genotype I was observed in all C. difficile of patients and environment. Genotypes III was not detected in any C. difficile isolates except in one of the acquired isolates. C. difficile is frequently transmitted among hospitalized patients, staff, and their hospital environment.
2014
Background and Aims: Clostridium difficile is an identified cause of antibioticassociated diarrhea, antibiotic-associated colitis, pseudomembranous colitis and nosocomial diarrhea. The objective of this survey was to determine molecular analysis of toxigenic Clostridium difficile isolates from hospital environment in Tehran tertiary medical centers. Materials and Methods: In this descriptive study, 100 hospital environmental specimens were collected. The specimens were cultured on a selective cycloserine cefoxitin fructose agar, and incubated in anaerobic conditions, at 37°C for 2 days. Clostridium difficile isolates were characterized by conventional biochemical tests. Bacterial cytotoxicity was assayed on tissue culture, and also all strains were typed by PCR ribotyping method. Results: Among toxigenic Clostridium difficile isolates, 6 isolates had the same PCR ribotyping patterns, and 11 isolates were typed in four different groups. Conclusion: Our findings showed that toxigenic ...
Frontiers in medicine, 2017
Clostridium difficile is an important cause of infectious colitis among hospitalized patients across the globe. The pathogenic potential of C. difficile in producing significant morbidity and mortality is mainly due to production of toxins A and B. The outbreaks of C. difficile infection (CDI) are due to changes in the genetic sequences of the organism. There is hardly any molecular study reported on the prevalent types of C. difficile strains in India. Toxinotyping and sequencing of locally circulating C. difficile isolates from patients presenting to our tertiary care center of North India were done. C. difficile strains (n = 174) isolated from 1,110 fecal samples from patients with suspected CDI were subjected to toxinotyping and partial sequencing of tcdA and tcdB genes. Comparison of nucleotide sequences with reference C. difficile 630 strain using BLAST was made and translated into corresponding amino acid sequences by ExPASy. Of 174 C. difficile isolates, 121 were toxigenic, ...
Anaerobe, 2017
This study aimed to characterize Clostridium difficile isolates cultured from stool samples of patients with C. difficile infection (CDI) and swabs from a medical environment in a gastroenterology centre in Tehran, Iran. A total of 158 samples (105 stool samples from hospitalized patients and 53 swabs from medical devices and the environment) were collected from January 2011 to August 2011 and investigated for the presence of C. difficile by direct anaerobic culture on a selective media for C. difficile. C. difficile isolates were further characterized by capillary electrophoresis (CE) ribotyping and toxin gene multiplex PCR. Of 158 samples, C. difficile was cultured in 19 of 105 stool samples (18%) and in 4 of 53 swabs (7.5%). C. difficile PCR ribotype (RT) 126 was the most common RT in the study (21.7%). Further RTs were: 001, 003, 014, 017, 029, 039, 081, 103 and 150. RTs 126, 001, 150 were cultured from both the stool samples and swabs of medical devices and the hospital environ...
Journal of Medical Microbiology, 2005
Clostridium difficile A þ B þ and A À B þ strains isolated from stool samples of patients with C. difficileassociated diarrhoea (CDAD) were selected from the University Hospital Warsaw collection. The binary-toxin genes cdtA and cdtB were detected by PCR in five of the 41 A þ B þ strains tested, but in none of the 17 A À B þ strains tested, giving 8 . 6 % prevalence (5/58) of binary-toxin-positive strains. All of the strains that were positive for binary-toxin genes were grouped into toxinotype IV, suggesting that in this institution toxinotype IV might dominate among the population of C. difficile with binary-toxin genes.
Journal of evolution of medical and dental sciences, 2015
BACKGROUND: Molecular methods for detection of toxigenic Clostridium difficile have been established in the developed countries though not very common in our country. AIMS: The study was intended to determine the presence of toxin A and toxin B genes of Clostridium difficile isolates by means of polymerase chain reaction (PCR) and analysis of clinical picture of the patients. MATERIALS AND METHODS: The prospective study was conducted in a tertiary care teaching hospital, South India from January 2012 to December 2014. Stool samples were collected consecutively from 563 in patients with diarrhoea from various wards. Clostridium difficile was isolated and identified by semi quantitative culture, latex agglutination and biochemical reactions. These isolates were then subjected to PCR for the detection of toxin A and toxin B genes. In addition, enzyme immunoassay was performed on stool samples for the detection of toxins A and B. The clinical spectrum of PCR positive patients was also analyzed. RESULTS: From 563 stool specimens, 113 (20.07%) Clostridium difficile isolates were grown by culture and identified by latex agglutination and biochemical reactions. Out of 113 isolates, 94 were subjected to PCR. 50 (53.19%) isolates out of 94 were found to be positive. Three toxigenic types obtained were A + B + , A-B + and A + Bwhich accounted for 6.38%, 42.55% and 4.26% respectively. A-Bisolates were 46.81%. 30 (26.55%) out of 113 stool samples (which were culture positive) was also enzyme immunoassay positive. 32 (64%) out of 50 PCR positive patients exhibited antibiotic usage (p<0.05) and 39(78%) revealed the presence of underlying illnesses/conditions (p<0.01). CONCLUSION: The study highlights the usefulness of PCR for detection of toxigenic Clostridium difficile and for determination of its molecular epidemiology.
Diagnosis of Clostridium difficile infection by toxigenic culture and PCR assay
Iranian Journal of Microbiology, 2018
Background and Objectives: Clostridium difficile is responsible for 15–25% of nosocomial antibiotic associated diarrhea (AAD) cases and all cases of pseudomembranous colitis. C. difficile has two major virulence factors, toxin A (enterotoxin) and toxin B (cytotoxin). The aim of this study was to determine the frequency of C. difficile strains in patients with diarrhea in Babol’ hospitals with toxigenic culture and PCR assay. Materials and Methods: One hundred stool specimens were taken from diarrheal patients in hospitals of the city of Babol. All patients had a history of antibiotic use. The samples were cultured on CCFA medium. In the next stage, toxigenic culture was performed for isolated C. difficile strains. Then, PCR assay was used to identify gdh, tcdA and tcdB genes among isolated C. difficile strains. Results: From the 100 stool samples, eight (8%) samples were positive in C. difficile culture. In toxigenic culture, two (2%) of these strains had cytopathic effects on Vero ...