engleski

Differentiation of infectious bursal disease virus strains by single strand conformational polymorphism analysis

Single-strand conformational polymorphism (SSCP) analysis of polymerase chain reaction (PCR) amplified products is a sensitive technique for rapid scanning of mutations occurring in single stranded DNAs. It is based on the fact that single stranded DNAs differing by a single base substitution may form different conformers. The conformational changes can be detected as the alterations in the electrophoretic mobility in non-denaturating polyacrylamide gels. The SSCP method was applied by us for differentiation of infectious bursal disease virus (IBDV) isolates. Field isolates were analyzed for variations in 422-bp fragment of VP2 gene. The single stranded DNAs, obtained by treatment of PCR products by alkali denaturing buffer (ADB), were electrophoretically analyzed in 8 % polyacrilamide gel. The SSCP analysis showed the difference between analyzed IBDV field isolates and confirmed our previous results of isolate differentiation carried out by restriction analysis using Cfo I, Sac I, Ssp I, Stu I and Taq I enzymes (Lojkić et al. (2003) Avian Dis. 47: 59).

infectious birsal disease virus; field isolates; single-stranded DNA; electrophoresis

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