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DNA of CVI 988 strain of Marek's disease virus quantified by Real-Time PCR in lung and bone marrow tissues after parenteral application to newly-hatched chicks

To determine the pathogenesis of Marek’s disease it is important to investigate kinetics of the causal virus in different tissues of chickens together with quantity of viral load. Previous research results confirmed that lungs are the place of viral entry in chicken organism, but kinetics in bone marrow was never analyzed nor compared with kinetics in other immunocompetent tissues. In this research kinetics of cell associated Marek’s disease virus (MDV), strain CVI 988, contained in Nobilis Rismavac vaccine applied parenteraly was analyzed. The same absolute quantification of mentioned virus in lungs and bone marrow was performed. Whole DNA was isolated from both tissues using commercial kit DNeasy Tissue Kit (Qiagen, USA). Using TaqMan probe and primers specific for the meq genome region of the MDV, Real-Time PCR was performed on Mx3005P® (Stratagene, USA) for DNA detection and kinetics in mentioned tissues, as well as for absolute quantification. Kinetics of the used virus did not differ from previous results for other immunocompetent organs after parenteral application of the same virus, or herpesvirus of turkey (HVT). On the other hand it differed from the kinetics of freeze dried HVT after being delivered by means of nebulisation. This is the first comparative description of MDV kinetics in lungs and bone marrow of chicks.

Marek’s disease; CVI 988; poultry; vaccination; kinetics; quantification; RealTime-PCR

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