Application of PCR for the detection of FMD viral RNA
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The polymerase chain reaction (PCR) method has been applied to the detection of FMD viral RNA in samples taken during the clinical stage of FMD from calf. Total RNA was extracted with guanidinum thiocyanate-phenol-chloroform method and reversety transcribed using AMV-reverse transcriptase. cDNA was used as a template for the amplification by PCR of the 672 bp of the VP1 coding sequence.The amplified fragment of cDNA was cloned in the phagemid pBS(+). The first DNA strand was sequenced and consistently an amino acid sequence was established. Comparison between VP1 amino acid sequence of FMDV types C and earlier described type O was performed.
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G. Paprocka, Zaklad Pryszczycy, Panstwowy Instytut Weterynaryjny, ul. Wodna 7, 98-220 Zdunska Wola, Poland