scholarly journals High Prevalence of Toxigenic and NontoxigenicClostridium difficileStrains in Malaysia

2018 ◽  
Vol 56 (6) ◽  
Author(s):  
Thomas V. Riley ◽  
Deirdre A. Collins ◽  
Rina Karunakaran ◽  
Maria Abdul Kahar ◽  
Ariza Adnan ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Southeast Asia ◽  
Kuala Lumpur ◽  
Content Type ◽  
Clinical Presentations ◽  
High Prevalence ◽  
Pcr Ribotyping ◽  
Severe Infections ◽  
Stool Specimens ◽  
Toxigenic Strains

ABSTRACTAccumulating evidence shows a high prevalence ofClostridium difficilein Southeast Asia associated with a range of clinical presentations. However, severe infections are rarely reported. We investigatedC. difficileinfection (CDI) across four hospitals in Kuala Lumpur and Kota Bharu, Malaysia. Enzyme immunoassays for glutamate dehydrogenase (GDH) and toxin A or B were performed on diarrheal stool specimens collected from patients in 2015 and 2016. Specimens were also cultured and isolates ofC. difficilecharacterized by PCR ribotyping and detection of toxin genes. In total, 437 specimens were collected and fecal toxin was detected in 3.0%. A further 16.2% of specimens were GDH positive and toxin negative. After culture, toxigenic strains were isolated from 10.3% and nontoxigenic strains from 12.4% of specimens. The most prevalent PCR ribotypes (RTs) were RT 017 (20.0%) and RT 043 (10.0%). The high prevalence of RT 017 and nontoxigenic strains in Malaysia and in neighboring Thailand and Indonesia suggests that they localize to the region of Southeast Asia, with an implication that they may mediate the burden of CDI in the region.

scholarly journals Cross-Sectional Study Reveals High Prevalence of Clostridium difficile Non-PCR Ribotype 078 Strains in Australian Veal Calves at Slaughter

2013 ◽  
Vol 79 (8) ◽  
pp. 2630-2635 ◽  
Author(s):  
Daniel R. Knight ◽  
Sara Thean ◽  
Papanin Putsathit ◽  
Stan Fenwick ◽  
Thomas V. Riley
Keyword(s):  
Clostridium Difficile ◽  
Cross Sectional Study ◽  
Sectional Study ◽  
Cross Sectional ◽  
Adult Cattle ◽  
Content Type ◽  
Retail Meats ◽  
High Prevalence ◽  
Pcr Ribotyping ◽  
Ribotype 078

ABSTRACTRecent reports in North America and Europe ofClostridium difficilebeing isolated from livestock and retail meats of bovine origin have raised concerns about the risk to public health. To assess the situation in Australia, we investigated the prevalence and genetic diversity ofC. difficilein adult cattle and calves at slaughter. Carcass washings, gastrointestinal contents, and feces were collected from abattoirs across five Australian states. Selective culture, toxin profiling, and PCR ribotyping were performed. The prevalence ofC. difficilewas 56% (203/360 samples) in feces from <7-day-old calves, 3.8% (1/26) in 2- to 6-month-old calves, and 1.8% (5/280) in adult cattle. Three PCR ribotypes (RTs), RT127, RT033, and RT126, predominated in <7-day-old calves and comprised 77.8% (158/203 samples) of isolates. RT056, which has not been reported in cattle before, was found in 16 <7-day-old calves (7.7%). Surprisingly, RT078 strains, which dominate production animal carriage studies in the Northern Hemisphere, were not isolated.

scholarly journals Nationwide Surveillance Study of Clostridium difficile in Australian Neonatal Pigs Shows High Prevalence and Heterogeneity of PCR Ribotypes

2014 ◽  
Vol 81 (1) ◽  
pp. 119-123 ◽  
Author(s):  
Daniel R. Knight ◽  
Michele M. Squire ◽  
Thomas V. Riley
Keyword(s):  
Clostridium Difficile ◽  
Enrichment Culture ◽  
Content Type ◽  
Neonatal Piglets ◽  
Neonatal Pigs ◽  
High Prevalence ◽  
Chromogenic Agar ◽  
Pcr Ribotyping ◽  
Nationwide Surveillance ◽  
Content Recovery

ABSTRACTClostridium difficileis an important enteric pathogen of humans and the cause of diarrhea and enteritis in neonatal pigs. Outside Australia, prevalence in piglets can be up to 73%, with a single PCR ribotype (RT), 078, predominating. We investigated the prevalence and genotype ofC. difficilein Australian pig herds. Rectal swabs (n= 229) were collected from piglets aged <7 days from 21 farms across Australia. Selective culture forC. difficilewas performed and isolates characterized by PCR for toxin genes and PCR ribotyping.C. difficilewas isolated from 52% of samples by direct culture on chromogenic agar and 67% by enrichment culture (P= 0.001). No association betweenC. difficilerecovery or genotype and diarrheic status of either farm or piglets was found. The majority (87%; 130/154) of isolates were toxigenic. Typing revealed 23 different RTs, several of which are known to cause disease in humans, including RT014, which was isolated most commonly (23%; 36/154). RT078 was not detected. This study shows that colonization of Australian neonatal piglets withC. difficileis widespread in the herds sampled.

scholarly journals First Environmental Investigation of Toxigenic Clostridium difficile at a Large Hospital in Bangladesh

2017 ◽  
Vol 4 (suppl_1) ◽  
pp. S406-S406
Author(s):  
M Jahangir Alam ◽  
Khurshida Begum ◽  
Tasnuva Rashid ◽  
Irtiza Hasan ◽  
Jacob McPherson ◽  
...  
Keyword(s):  
Developing Countries ◽  
Clostridium Difficile ◽  
Enrichment Culture ◽  
Large Hospital ◽  
Environmental Investigation ◽  
Fluorescent Pcr ◽  
High Prevalence ◽  
Pcr Ribotyping ◽  
Developed World ◽  
Culture Positive

Abstract Background Toxigenic Clostridium difficile is the most common cause of infectious diarrhea in hospitalized patients in the developed world and an emerging pathogen in developing countries due to increased use of broad-spectrum antibiotics. Although likely ubiquitous worldwide, the prevalence of toxigenic C. difficile spores in the hospital environs of developing countries is poorly understood. The objectives of the study are to isolate and characterize C. difficile from the hospital environs of a large hospitals in Dhaka, Bangladesh. Methods As part of our environmental surveillance effort, we collected 330 shoe-bottom swab samples from hospital employees, patients, and visitors inside of a large hospital in Dhaka, Bangladesh. Samples were analyzed for C. difficile using anaerobic enrichment culture and molecular methods. Suspected colonies from cycloserine cefoxitin fructose agar (CCFA) plates were identified by PCR (tcdA, tcdB, cdtA, cdtB and tpi genes) and strain typed using fluorescent PCR ribotyping, and MLVA methods. Results A total 149 of 333 (44.7%) shoe-bottom swab samples were culture positive for C. difficile of which 19.8% samples were toxigenic (tcdA and tcdB) C. difficile. A total of 11 distinct ribotypes were identified from 58 toxigenic C. difficile isolates tested. Predominant ribotypes were F053-163 (24.1%), F017 (20.7%), F106 (19.0%), F014-020 (17.2%). Other ribotypes were R001, F005, F010, F018, F054, F216, and FP407. No R027 and R078 C. difficile isolated. A broad MLVA diversity has been seen among the tested strains. Conclusion We identified a high prevalence of toxigenic C. difficile with diverse ribotypes from hospital environmental shoe-bottom swabs in Bangladesh. This is the first hospital environmental report of C. difficile from Bangladesh. Disclosures All authors: No reported disclosures.

scholarly journals Evaluation of Portability and Cost of a Fluorescent PCR Ribotyping Protocol for Clostridium difficile Epidemiology

2015 ◽  
Vol 53 (4) ◽  
pp. 1192-1197 ◽  
Author(s):  
Jonathan N. V. Martinson ◽  
Susan Broadaway ◽  
Egan Lohman ◽  
Christina Johnson ◽  
M. Jahangir Alam ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Low Cost ◽  
Cost Effective ◽  
Pcr Primers ◽  
The United States ◽  
Content Type ◽  
Fluorescent Pcr ◽  
Address Data ◽  
Pcr Ribotyping ◽  
Data Portability

Clostridium difficileis the most commonly identified pathogen among health care-associated infections in the United States. There is a need for accurate and low-cost typing tools that produce comparable data across studies (i.e., portable data) to help characterize isolates during epidemiologic investigations ofC. difficileoutbreaks and sporadic cases of disease. The most popularC. difficile-typing technique is PCR ribotyping, and we previously developed methods using fluorescent PCR primers and amplicon sizing on a Sanger-style sequencer to generate fluorescent PCR ribotyping data. This technique has been used to characterize tens of thousands ofC. difficileisolates from cases of disease. Here, we present validation of a protocol for the cost-effective generation of fluorescent PCR ribotyping data. A key component of this protocol is the ability to accurately identify PCR ribotypes against an online database (http://walklab.rcg.montana.edu) at no cost. We present results from a blinded multicenter study to address data portability across four different laboratories and three different sequencing centers. Our standardized protocol and centralized database for typing ofC. difficilepathogens will increase comparability between studies so that important epidemiologic linkages between cases of disease and patterns of emergence can be rapidly identified.

scholarly journals Typing of Clostridium difficile isolates endemic in Japan by sequencing of slpA and its application to direct typing

2010 ◽  
Vol 59 (5) ◽  
pp. 556-562 ◽  
Author(s):  
Haru Kato ◽  
Hideaki Kato ◽  
Yoichiro Ito ◽  
Takayuki Akahane ◽  
Sayuri Izumida ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Protein A ◽  
Sequence Type ◽  
Typing System ◽  
Pcr Ribotyping ◽  
Stool Specimens ◽  
Encoding Gene ◽  
Ribotype 078 ◽  
Toxic Culture ◽  
Pcr Ribotype 027

A typing system for Clostridium difficile using sequencing of the surface-layer protein A encoding gene (slpA) was evaluated and used to analyse clinical isolates in Japan. A total of 160 stool specimens from symptomatic patients in Japan was examined and 87 C. difficile isolates were recovered. slpA sequence typing was found to have reliable typability and discriminatory power in comparison with PCR ribotyping, and the typing results were highly reproducible and comparable. slpA sequence typing was used to type C. difficile in DNA extracted directly from stool specimens. Among the 90 stool specimens in which direct typing results were obtained, 77 specimens were positive for C. difficile culture, and typing results from isolated strains agreed with those from direct typing in all 77 specimens. The slpA sequence type smz was dominant at all four hospitals examined, and this endemic type was detected by culture and/or direct typing in 61 (62 %) of 99 stool specimens positive for toxic culture and/or direct slpA sequence typing. Comparison of epidemic strains reported throughout the world revealed one isolate identified as slpA sequence type gc8, which was found to correspond to PCR ribotype 027 (BI/NAP1/027), whereas no isolates were found with the slpA gene identical to that of PCR ribotype 078 strain. slpA sequence typing is valuable for comparison of C. difficile strains epidemic in diverse areas because the typing results are reproducible and can easily be shared. In addition, slpA sequence typing could be applied to direct typing without culture.

scholarly journals High Prevalence of Clostridium difficile in Home Gardens in Western Australia

2020 ◽  
Vol 87 (1) ◽  
Author(s):  
Nirajmohan Shivaperumal ◽  
Barbara J. Chang ◽  
Thomas V. Riley
Keyword(s):  
Risk Factors ◽  
Clostridium Difficile ◽  
Western Australia ◽  
Brain Heart Infusion ◽  
Home Gardens ◽  
Home Garden ◽  
Toxin Gene ◽  
Content Type ◽  
High Prevalence ◽  
Shoe Soles

ABSTRACT In recent years, community-associated Clostridium difficile infection (CA-CDI) has emerged as a significant health problem, accounting for ∼50% of all CDI cases. We hypothesized that the home garden environment could contribute to the dissemination of C. difficile spores in the community and investigated 23 homes in 22 suburbs of Perth, Western Australia. We identified a high prevalence of toxigenic C. difficile in this environment. In total, 97 samples consisting of soil (n = 48), compost (n = 15), manure (n = 12), and shoe sole swabs (n = 22) were collected. All samples were cultured anaerobically on C. difficile ChromID agar and enriched in brain heart infusion broth, and isolates were characterized by toxin gene PCR and PCR ribotyping. Two-thirds (67%; 95% confidence interval [CI], 57 to 76%) of home garden samples, including 79% (95% CI, 68 to 91%) of soil, 67% (95% CI, 43 to 90%) of compost, 83% (95% CI, 62% to 100%) of manure, and 32% (95% CI, 12 to 51%) of shoe sole samples, contained C. difficile. Of 87 isolates, 38% (95% CI, 28 to 48%) were toxigenic, and 26 PCR ribotypes (RTs), 5 of which were novel, were identified. The toxigenic C. difficile strain RT014/020 was the most prevalent RT. Interestingly, 19 esculin hydrolysis-negative strains giving white colonies were identified on C. difficile ChromID agar, 5 of which were novel toxigenic RTs that produced only toxin A. Clearly, there is the potential for transmission of C. difficile in the community due to the contamination of home gardens. Our findings highlight the importance of a “One Health” approach to dealing with CDI. IMPORTANCE Recently, community-associated Clostridium difficile infection (CA-CDI) has emerged as a significant problem, accounting for ∼50% of all CDI cases and reported to affect a younger population without traditional risk factors. Possible sources of CA-CDI are soil, food, and water contaminated by animal feces, and recent reports show overlapping ribotypes of C. difficile in animals, humans, and the environment; however, the epidemiology of CA-CDI and related risk factors need to be better understood. Our research aimed to determine the prevalence of C. difficile in home gardens and on the shoe soles of homeowners in Perth, Western Australia. There were high rates of contamination with C. difficile in gardens, and some of the ribotypes identified had been isolated from human cases of CDI in Western Australia. This study shows that home gardens and shoes may be a source of C. difficile in CA-CDI.

scholarly journals Evaluation of the QiagenartusC. difficile QS-RGQ Kit for Detection of Clostridium difficile Toxins A and B in Clinical Stool Specimens

2015 ◽  
Vol 53 (6) ◽  
pp. 1942-1944 ◽  
Author(s):  
Nathalie Jazmati ◽  
Pia Wiegel ◽  
Božica Ličanin ◽  
Georg Plum
Keyword(s):  
Nucleic Acid ◽  
Clostridium Difficile ◽  
Positive Predictive Value ◽  
Negative Predictive Value ◽  
Predictive Value ◽  
Nucleic Acid Amplification Test ◽  
Nucleic Acid Amplification ◽  
Content Type ◽  
Stool Specimens ◽  
Sensitivity Specificity

We compared the QiagenartusC. difficile QS-RGQ kit, a new nucleic acid amplification test for the detection ofClostridium difficiletoxins in stool specimens, with the Cepheid XpertC. difficiletest. The sensitivity, specificity, positive predictive value, and negative predictive value for the QiagenartusC. difficile QS-RGQ test were 100%, 89.5%, 60.9%, and 100%, and those for the Cepheid XpertC. difficiletest were 100%, 90%, 62.2%, and 100%, respectively.

scholarly journals Coinfection and Emergence of Rifamycin Resistance during a Recurrent Clostridium difficile Infection

2016 ◽  
Vol 54 (11) ◽  
pp. 2689-2694 ◽  
Author(s):  
Emma C. Stevenson ◽  
Giles A. Major ◽  
Robin C. Spiller ◽  
Sarah A. Kuehne ◽  
Nigel P. Minton
Keyword(s):  
Clostridium Difficile ◽  
Recurrent Infection ◽  
Disease Symptoms ◽  
Content Type ◽  
Recurrent Clostridium Difficile Infection ◽  
High Incidence ◽  
Life Threatening ◽  
Pcr Ribotyping ◽  
Stool Samples ◽  
Health Care Associated Infection

Clostridium difficile ( Peptoclostridium difficile ) is a common health care-associated infection with a disproportionately high incidence in elderly patients. Disease symptoms range from mild diarrhea to life-threatening pseudomembranous colitis. Around 20% of patients may suffer recurrent disease, which often requires rehospitalization of patients. C. difficile was isolated from stool samples from a patient with two recurrent C. difficile infections. PCR ribotyping, whole-genome sequencing, and phenotypic assays were used to characterize these isolates. Genotypic and phenotypic screening of C. difficile isolates revealed multiple PCR ribotypes present and the emergence of rifamycin resistance during the infection cycle. Understanding both the clinical and bacterial factors that contribute to the course of recurrent infection could inform strategies to reduce recurrence. (This study has been registered at ClinicalTrials.gov under registration no. NCT01670149.)

scholarly journals DNA Microarray-Based PCR Ribotyping of Clostridium difficile

2014 ◽  
Vol 53 (2) ◽  
pp. 433-442 ◽  
Author(s):  
Alexander Schneeberg ◽  
Ralf Ehricht ◽  
Peter Slickers ◽  
Vico Baier ◽  
Heinrich Neubauer ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Dna Microarray ◽  
Genetic Relatedness ◽  
Intergenic Spacer ◽  
Matrix Analysis ◽  
Content Type ◽  
Intergenic Spacer Region ◽  
Hybridization Probes ◽  
Pcr Ribotyping ◽  
Set Up

This study presents a DNA microarray-based assay for fast and simple PCR ribotyping ofClostridium difficilestrains. Hybridization probes were designed to query the modularly structured intergenic spacer region (ISR), which is also the template for conventional and PCR ribotyping with subsequent capillary gel electrophoresis (seq-PCR) ribotyping. The probes were derived from sequences available in GenBank as well as from theoretical ISR module combinations. A database of reference hybridization patterns was set up from a collection of 142 well-characterizedC. difficileisolates representing 48 seq-PCR ribotypes. The reference hybridization patterns calculated by the arithmetic mean were compared using a similarity matrix analysis. The 48 investigated seq-PCR ribotypes revealed 27 array profiles that were clearly distinguishable. The most frequent human-pathogenic ribotypes 001, 014/020, 027, and 078/126 were discriminated by the microarray.C. difficilestrains related to 078/126 (033, 045/FLI01, 078, 126, 126/FLI01, 413, 413/FLI01, 598, 620, 652, and 660) and 014/020 (014, 020, and 449) showed similar hybridization patterns, confirming their genetic relatedness, which was previously reported. A panel of 50C. difficilefield isolates was tested by seq-PCR ribotyping and the DNA microarray-based assay in parallel. Taking into account that the current version of the microarray does not discriminate some closely related seq-PCR ribotypes, all isolates were typed correctly. Moreover, seq-PCR ribotypes without reference profiles available in the database (ribotype 009 and 5 new types) were correctly recognized as new ribotypes, confirming the performance and expansion potential of the microarray.

scholarly journals Molecular Epidemiology of Clostridium difficile Infection in Hospitalized Patients in Eastern China

2016 ◽  
Vol 55 (3) ◽  
pp. 801-810 ◽  
Author(s):  
Dazhi Jin ◽  
Yun Luo ◽  
Chen Huang ◽  
Jian Cai ◽  
Julian Ye ◽  
...  
Keyword(s):  
Risk Factors ◽  
Clostridium Difficile ◽  
Antimicrobial Resistance ◽  
Severity Score ◽  
Eastern China ◽  
Developed Countries ◽  
Hospitalized Patients ◽  
Chinese Patients ◽  
Content Type ◽  
Stool Specimens

ABSTRACT Few studies on risk factors for and transmission of Clostridium difficile infection (CDI) in China have been reported. A cross-sectional study was conducted for 3 years in eastern China. Consecutive stool specimens from hospitalized patients with diarrhea were cultured for C. difficile. C. difficile isolates from these patients then were analyzed for toxin genes, genotypes, and antimicrobial resistance. A severity score for the CDI in each patient was determined by a blinded review of the medical record, and these scores ranged from 1 to 6. A total of 397 out of 3,953 patients (10.0%) with diarrhea were found to have CDI. Severity of CDI was mild to moderate, and the average (± standard deviation) severity score was 2.61 ± 1.01. C. difficile was isolated from stool specimens in 432 (10.9%) of all the patients who had diarrhea. C. difficile genotypes were determined by multilocus sequence analysis and PCR ribotyping; sequence type 37 (ST37)/ribotype 017 (RT017) ( n = 68, 16.5%) was the dominant genotype. Eleven patients (16.2%) with this genotype had a CDI severity score of 5. Overall, three RTs and four STs were predominant; these genotypes were associated with significantly different antimicrobial resistance patterns in comparison to all genotypes (χ 2 = 79.56 to 97.76; P < 0.001). Independent risk factors associated with CDI included age greater than 55 years (odds ratio [95% confidence interval], 26.80 [18.76 to 38.29]), previous hospitalization (12.42 [8.85 to 17.43]), previous antimicrobial treatment within 8 weeks (150.56 [73.11 to 310.06]), hospital stay more than 3 days before sampling (2.34 [1.71 to 3.22]), undergoing chemotherapy (3.31 [2.22 to 4.92]), and undergoing abdominal surgery (4.82 [3.54 to 6.55]). CDI is clearly a problem in eastern China and has a prevalence of 10.0% in hospitalized patients. Among risk factors for CDI, the advanced age threshold was younger for Chinese patients than that reported for patients in developed countries.

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