Molecular characterization of clostridium difficile isolates by capillary gel electrophoresis-based PCR ribotyping

2012 ◽  
Author(s):  
Ching-to Lam
Keyword(s):  
Clostridium Difficile ◽  
Molecular Characterization ◽  
Gel Electrophoresis ◽  
Capillary Gel Electrophoresis ◽  
Pcr Ribotyping

scholarly journals Characterization of Clostridium difficile isolates using capillary gel electrophoresis-based PCR ribotyping

2008 ◽  
Vol 57 (11) ◽  
pp. 1377-1382 ◽  
Author(s):  
A. Indra ◽  
S. Huhulescu ◽  
M. Schneeweis ◽  
P. Hasenberger ◽  
S. Kernbichler ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Gel Electrophoresis ◽  
Variable Number Tandem Repeat ◽  
Variable Number ◽  
Web Based ◽  
Healthcare Facilities ◽  
Capillary Gel Electrophoresis ◽  
Hands On ◽  
Pcr Ribotyping ◽  
Reference Strains

We have developed a Clostridium difficile PCR ribotyping method based on capillary gel electrophoresis and have compared it with conventional PCR ribotyping. A total of 146 C. difficile isolates were studied: five isolates were reference strains (PCR ribotypes 001, 014, 017, 027 and 053); 141 were clinical isolates comprising 39 Austrian PCR ribotypes collected in the period 2006–2007 at 25 Austrian healthcare facilities. Capillary gel electrophoresis yielded up to 11 fragments per isolate and 47 ribotype patterns. All but one of the five PCR ribotypes of reference strains were clearly reflected in the chromatograms of capillary-based typing. Capillary gel electrophoresis divided 24 isolates belonging to PCR ribotype type 014 into seven subgroups, whereas subtyping the same isolates using multiple-locus variable-number tandem-repeat analysis yielded three unrelated subgroups, without obvious correlation to sr subgroups. Using a web-based software program (http://webribo.ages.at), we were able to correctly identify these 014 isolates by simply allocating the seven subgroup patterns to one ribotype, i.e. to PCR ribotype 014. We consider capillary gel electrophoresis-based PCR ribotyping to be a way of overcoming the problems associated with inter-laboratory comparisons of typing results, while at the same time substantially diminishing the hands-on time for PCR ribotyping.

scholarly journals Characterization of clinical Clostridium difficile isolates by PCR ribotyping and detection of toxin genes in Austria, 2006–2007

2008 ◽  
Vol 57 (6) ◽  
pp. 702-708 ◽  
Author(s):  
A. Indra ◽  
D. Schmid ◽  
S. Huhulescu ◽  
M. Hell ◽  
R. Gattringer ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Pseudomembranous Colitis ◽  
Antimicrobial Agents ◽  
Toxic Megacolon ◽  
Health Care Facilities ◽  
Reference Laboratory ◽  
Specimen Collection ◽  
Pcr Ribotyping ◽  
High Level

In order to assess the lethality of Clostridium difficile-associated disease (CDAD) and the PCR ribotypes prevalent in Austria, the Austrian Agency for Health and Food Safety requested isolates of C. difficile from patients in a structured but arbitrary sampling scheme. In the allocated period from February 2006 to January 2007, local hospital laboratories within each of the nine provinces were asked to submit C. difficile isolates from at least ten cases of CDAD. Confirmation of species identification, toxin detection, susceptibility testing against four antimicrobial agents and typing using a PCR ribotyping method were performed at the reference laboratory. In total, 149 isolates of putative C. difficile were submitted, from which 142 were included for study. Antimicrobial susceptibility patterns revealed resistance to clindamycin in 57 % and high-level resistance to moxifloxacin in 38 % of isolates tested. CDAD manifested as diarrhoea (including eight cases of bloody diarrhoea) in 126 cases (88.7 %), as pseudomembranous colitis in 15 cases (10.6 %) and as toxic megacolon in one case. Twelve of the 142 patients died within 30 days of specimen collection (8.45 % lethality). A lethal outcome occurred in 2/15 cases (13.3 %) when pseudomembranous colitis was present and in 10/126 cases (7.9 %) in the absence of pseudomembranous colitis or toxic megacolon. Among the 142 isolates from 25 health-care facilities, 41 PCR ribotype patterns were found. The most frequent ribotypes were AI-5 (including six lethal cases out of 26 patients), 014 (two out of 24) and 053 (one out of 24). The typing patterns demonstrated the occurrence of clusters in hospitals.

Molecular characterization of Clostridium difficile isolated from carriage and association of its pathogenicity to prevalent toxic genes

2018 ◽  
Vol 120 ◽  
pp. 1-7
Author(s):  
Abdulwahab Ali Abuderman ◽  
Ayesha Mateen ◽  
Rabbani Syed ◽  
Mustafa sawsan aloahd
Keyword(s):  
Clostridium Difficile ◽  
Molecular Characterization

scholarly journals Genotypic Characterization of Coagulase-negative Staphylococci Isolated from Sheep Milk in Slovakia

2010 ◽  
Vol 79 (2) ◽  
pp. 269-275 ◽  
Author(s):  
Ivana Pilipčincová ◽  
Mangesh Bhide ◽  
Eva Dudriková ◽  
Milan Trávniček
Keyword(s):  
Molecular Characterization ◽  
Gel Electrophoresis ◽  
Biochemical Characterization ◽  
Pulse Field ◽  
Coagulase Negative Staphylococci ◽  
Pulse Field Gel Electrophoresis ◽  
Milk Samples ◽  
Sheep Milk ◽  
Epidemiological Situation

Hitherto very few reports are available presenting identification and molecular characterization of the coagulase negative staphylococci (CNS) from sheep milk in the subclinical stage of mastitis. Furthermore, very scanty data are available on the epidemiological status of CNS in different Slovak provinces. Milk samples from 54 sheep farms located in eastern Slovak region were screened. A total 240 CNS were identified with series of biochemical testes (STAPH-API) and subjected further for genotyping with the help of pulse field gel electrophoresis (PFGE). The most frequently occurring CNS species according the biochemical characterization were:S. epidermidis(36.3 %),S. caprae(21.3 %),S. hominis(6.6 %),S. chromogenes(6.3 %),S. xylosus(5.8 %),S. warneri(5.0 %) andS. capitis(4.6 %). Further PFGE-based characterization of these isolates revealed six pulsotypes of theS. epidermidis, two ofS. caprae, three ofS. chromogenes, nine ofS. hominis, five ofS. capitisand seven ofS. xylosus. These results contribute to knowledge of the epidemiological situation of the CNS from the subclinical form of mastitis in Slovakia.

Molecular characterization of moxifloxacin resistance from Canadian Clostridium difficile clinical isolates

2010 ◽  
Vol 66 (4) ◽  
pp. 419-424 ◽  
Author(s):  
Andrew Walkty ◽  
David A. Boyd ◽  
Denise Gravel ◽  
Jim Hutchinson ◽  
Allison McGeer ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Molecular Characterization ◽  
Clinical Isolates

scholarly journals Molecular Characterization of Glycopeptide-Resistant Enterococcus faecium Isolates from Portuguese Hospitals

2005 ◽  
Vol 49 (7) ◽  
pp. 3073-3079 ◽  
Author(s):  
Carla Novais ◽  
João C. Sousa ◽  
Teresa M. Coque ◽  
Luísa V. Peixe
Keyword(s):  
Molecular Characterization ◽  
Enterococcus Faecium ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field ◽  
Vancomycin Resistant ◽  
Geographical Locations

ABSTRACT Fifty-one pulsed-field gel electrophoresis types and 17 Tn1546 variants were identified among 101 Enterococcus faecium isolates recovered in three distant Portuguese hospitals. Intra- and interhospital dissemination of specific strains and Tn1546 types was detected, which might largely contribute to the endemicity of vancomycin-resistant E. faecium in Portuguese hospitals, as happened previously in other geographical locations.

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