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[Identification of the genome of a picornavirus isolate using Random-PCR]
Objective:
To identify the genome of an enterovirus isolate (XJ90).
Methods:
The double-stranded cDNA of isolated virus was synthesized with anchored-random primer and was amplified with anchor primer; the PCR products were cloned into pGEM-T vector for sequencing and analysis.
Results:
Analysis on agarose gel electrophoresis of the Random-PCR products showed a typical smear; 11 randomly selected clones were all homologous to the members of enteroviruses and the homology rate over 80%; the 11 clones were distributed evenly along the whole genome and they assembled 4 contigs totally 2 261 bp long which covered 30% of the genome. The homology of the fourth contig and coxsackie virus B6 was 97.3% at amino acids level. The virus gene was specifically amplified with primers derived from the sequenced clones.
Conclusions:
The picornavirus isolate XJ90 is a member of enteroviruses.