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Ages-Webribo a new deterministic identification algorithm for the characterization of Clostridium difficile isolates by capillary gel electrophoresis-based PCR-ribotyping

Abstract number: P1715

Indra A., Blaschitz M., Schneewe M., Huhulescu S., Kernbichler S., Fiedler A., Allerberger F.

Objectives:Clostridium difficile is a frequently identified cause of hospital-acquired infections. Identification of newly emerging hypervirulent C. difficile strains – like 027, 078 and 053 is mostly done by PCR-ribotyping. Although ribotyping is widely used the method lacks an interlaboratory interchangeable format. In order to overcome capillary-sequencer-dependent result variations of different types of sequencing machines a newly developed deterministic identification algorithm – AGES-WEBRIBO (webribo.ages.at) – was applied for capillary gel electrophoresis based Clostridium difficile-PCR ribotyping.

Methods: 2600 isolates collected between the years 2006 to 2009 at Austrian healthcare facilities were primarily analysed with an Applied Biosystems (ABI) 310 Genetic analyzer yielding 303 different PCR-ribotypes. 65 of the 303 ribotypes were randomly chosen and retested with on an ABI 3130 Genetic Analyzer. Quality control of data gained was done using the AGES-WEBRIBO internet platform.

Results: There was no increase in discrimatory power using different sequencer machines, although there was better narrow double-peak resolution with the ABI-3130 system. Three (4.5%) of the 65 on the ABI-3130 sequencer examined samples showed no correct identification after analysing the data with AGES-WEBRIBO.

Conclusion: 95.5% correct identifications of C. difficile ribotypes were achieved between different ABI-Sequencer models with the newly developed deterministic-data-analysis approach (AGES-WEBRIBO). This result clearly shows that every user is able to determine even rare PCR-ribotypes without a respective reference strain at hand. Further tests will show the ability of AGES-WEBRIBO (webribo.ages.at) to correctly identify data gained with capilliary sequencer models from other suppliers than ABI. We consider the updated version of AGES-WEBRIBO in combination with capillary gel electrophoresis based PCR ribotyping to be a universal tool for inter-laboratory comparability of C. difficile ribotyping.

Session Details

Date: 10/04/2010
Time: 00:00-00:00
Session name: Abstracts 20th European Congress of Clinical Microbiology and Infectious Diseases
Subject:
Location: Vienna, Austria, 10 - 13 April 2010
Presentation type:
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