Using real-time Polymerase Chain Reaction (PCR) for detection of agents in the eprimental sepsis models

Sepsis is a disease that cause serious problems in the treatment and thus it has high mortality rates. Early diagnosis of sepsis is worth of research, in order to provide rapid diagnosis. PCR methodology is used for early and prompt diagnosis of sepsis.In our study, Real-Time PCR was used for the detection of infectious agents individually or in polymicrobial form.In this study 14 rabbits were used and divided into four groups, as; A, B, C and healthy control groups. C. albicans ATCC 10231 and A. fumigatus ATCC 204305 were injected into A group rabbits, while E. coli ATCC 10536 and S. aureus ATCC 6538 were injected into B group rabbits. Injections were performed by the ear ven. Equal proportion of fungi and bacteria suspension was injected into C group of rabbits. None of the microorganism injected into healthy controls.Blood and tissue samples were cultured and were found positive in all working groups. DNA samples were extracted from blood and tissue samples of the rabbits.Using specific primers and probes, the presence of fungal and bacterial sepsis was confirmed by TaqMan PCR. An additional multiplex PCR assay was performed using SYBR green and melting curve analysis.Pathogens were detected in most of the samples, while control groups were found negative.Molecular methods that used in the diagnosis of sepsis when compared with blood culture, can be provide more rapid results, specificity and sensitivity of these tests are high.Keywords: Bacteria, Fungi, Sepsis, Real-Time PCR, SYBR green, TaqMan