Application of Molecularbiological Methods in the Toxin-typing of Clostridium difficile

Abstract

Clostridium difficile is both a member of the normal human flora and a human pathogen with clinical significance, a causative agent of Clostridium difficile-associated diarrhoea. Determination of the strains and related information about their virulence is very important for the correct diagnosis and therapy of patients, the prophylaxis and the control of the spreading of the infection in society. The precise phenotyping of this organism is complicated because of the relatively low sensitivity of the classical microbiological methods and a low vitality of the anaerobe Clostridium difficile in an atmosphere of oxygen. The methods developed in the last decade for PCR-techniques directly from clinical samples have specific disadvantages.That was why we genotyped C. difficile using primers for the genes tcdA, tcdB and tcdC for a direct DNA isolation from pure culture suitable for work in our lab.