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1 Bulletin of the Veterinary Institute in Puławy

1 2001

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Polymerase chain reaction for the differentiation of Marek's disease virus strains

100%

Kozdrun W. , Samorek-Salamonowicz E. , Czekaj H.

tom 45

nr 1

The aim of this study was an attempt to apply PCR for diagnosis of Marek's disease. Eighteen field Marek's disease virus strains and 4 standard strains: virulent HPRS₁₆ strain and attenuated CVI 988 strain (serotype 1 - MDV-1), apathogenic SB 1strain (serotype 2 - MDV-2) and non - pathogenic turkeys HVT FC 126 strain (serotype 3 MDV-3) were used. Chicken anaemia virus (CAV) and infectious laryngotracheitis virus (ILTV) were used as control of PCR. The virus strains were cultured in CEF and total DNA was isolated by A@A Biotechnology kit. Three pairs of primers were used: for gene A of serotype 1, for 132 bp sequence of serotype 1 and for gene A of serotype 3. PCR was carried out under the following conditions: 94°C - 1 min (denaturation), 55°C - 30 s (annealing), 72°C -30 s (elongation). PCR products were analysed by electrophoresis in 2% agarose gel at 100 V/h. The PCR product of 314 bp was obtained from all field strains and HPRS₁₆ standard after the primers for gene A of serotype 1 were used. After the primers for 132 bp sequence of serotype 1 were used, 434 bp fragment for 19 DNA samples derivied from field strains and standard HPRS₁₆ strain and 1033 bp fragment from attenuated CVI 988 strain were obtained. Application of primers for gene A serotype 3 allowed the detection of 436 bp product only from non-pathogenic HVT FC 126 strain (serotype 3). These results indicated that the PCR technique can be used for the differentiation of MDV strains.