DETECTION OF THE PORCINE CYTOKINES MRNA LEVEL BY REAL-TIME RT-PCR
African swine fever virus (ASFV) is the etiological agent of a contagious and often lethal disease of domestic pigs that has significant economic consequences for the swine industry. The control of African Swine Fever (ASF) has been hampered by the unavailability of vaccines. The ASFV strains have different genetic and biological characteristics, degree of pathogenicity. The immune response with ASF significantly depends of the strain of the virus. The cytokine status is one of the most important characteristics of an immune response. For a better understanding of the functional properties of immune cells the analysis of cytokine profiles of animals infected with different ASFV strains is of fundamental importance. Reverse transcription polymerase chain reaction (RT-PCR) is widely applied to quantify the expression of cytokines in cells from various sources. The real-time PCR methods permit the sensitive detection and accurate quantification of cytokine mRNAs, which are sometimes expressed at very low levels in the target cells. An increase in the level of proinflammatory cytokines, such as TNF-a, interferons and chemokines, probably plays an important role in the pathology of ASF. We have established an easy real-time PCR assay, which allows the precise quantifica tion of changes in the expression level of 2 porcine pro-inflammatory cytokines (IFN-y and TNF-a), and 2 housekeeping genes.