The detection of Yersinia enterocolitica in biological samples by RT-PCR technique.

Abstract

The research has been performed in order to optimize Yersinia enterocolitica diagnosis from blood samples and other biological samples from different human clinical cases. Following this objective, the general technique of collecting and processing the samples was adapted and optimized, raising the rapidity of diagnosis. There have been artificially contaminated different types of samples with Yersinia enterocolitica stems, checking the efficiency and accuracy of optimized method. In finally was developed a methodology of Yersinia identification from naturally contaminated biological samples. As a target-gene there has been used the chromosome gene ail, this being present in the pathogenic strains of Y. enterocolitica. The sample was marked at the 5’ end with a report dyeing solution and at the 3’ end with an extinction one. The reading was performed at the RT-PCR, with a work protocol consisting of 2 stages, including 45 cycles. This qualitative method demonstrated some specific results with high sensitivity, positive ail gene detection rate in Y. enterocolitica in 267 blood samples and other type of biological products, being of 82 % using PCR and 56 % using culture methods.