[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.
Du
G
G
Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
Pan
W
W
Chen
W
W
Wang
H T
HT
D000595:Amino Acid Sequence; D004770:Enterovirus / Q000235:genetics* / Q000302:isolation & purification*; D005243:Feces / Q000821:virology; D016751:Hepatitis E / Q000821:virology; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods