The Detection of Yersinia enterocolitica in Biological Samples by RTPCR Technique

摘要

The research has been performed in order to optimize Yersinia enterocoliticadiagnosis 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 adaptedand optimized, raising the rapidity of diagnosis. There have been artificially contaminated differenttypes of samples with Yersinia enterocolitica stems, checking the efficiency and accuracy of optimizedmethod. In finally was developed a methodology of Yersinia identification from naturallycontaminated biological samples. As a target-gene there has been used the chromosome gene ail, thisbeing present in the pathogenic strains of Y. enterocolitica. The sample was marked at the 5 end witha report dyeing solution and at the 3 end with an extinction one. The reading was performed at theRT-PCR, with a work protocol consisting of 2 stages, including 45 cycles. This qualitative methoddemonstrated 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 PCRand 56 % using culture methods.