Original scientific paper
THE CALCULATION OF RELATIVE QUANTITIES MRNA FOR CHICKEN IFN-γ STIMULATED BY LA SOTA STRAIN OF NEWCASTLE DISEASE VIRUS
Mirta Balenović
; Hrvatski veterinarski institut, Centar za peradarstvo, Heinzelova 55, Zagreb, Hrvatska
V. Savić
; Hrvatski veterinarski institut, Centar za peradarstvo, Heinzelova 55, Zagreb, Hrvatska
Anamaria Ekert Kabalin
; Zavod za stočarstvo, Veterinarski fakultet Sveučilišta u Zagrebu, Heinzelova 55, Zagreb, Hrvatska
L. Jurinović
; Hrvatski veterinarski institut, Centar za peradarstvo, Heinzelova 55, Zagreb, Hrvatska
Abstract
One of significant prerequisites for successful intensive poultry production is healthy and immune competent poultry. Therefore, an interest for understanding the role of cell immunity has increased, that is, for the role of immune cells and molecules involved in the cell immunity with the aim of achieving better immune response. By developing molecular methods, it is possible to monitor the kinetics of immune molecules through a time period to the stimulation with immunogen. With the development of molecular methods, the monitoring of molecule synthesis dynamics after stimulation with immunogen became possible. In the study, the dynamics of interferon γ (IFN-γ) synthesis in the spleens of chicken embryos, after inoculation with the Newcastle disease virus of La Sota strain was determined. Standard curve was prepared in ten-fold dilutions of our gained positive control mRNA for IFN-γ. Through comparable reactions of the product, the number of copies in the sample was estimated, from the value of cycle threshold, that is, the cycle in which reporter colour was changed. Through the gained results for ten-fold dilution series, with mathematical calculation, a standard exponential curve was determined as well as an equation of mRNA quantity for IFN-γ in ten-fold dilution series of our positive control. According to the results of the signal occurrence in ten-fold dilution series of the known standard in which reporter dye was changed in real time reverse transcription - polymerase chain reaction method (real-time RT-PCR), with mathematical calculations, an adjusted exponential curve and an equation was calculated which best describe the gained values.
Keywords
Interferon γ; Newcastle disease; standard curve; exponential equation
Hrčak ID:
41285
URI
Publication date:
15.8.2009.
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