Use of denaturing gradient gel electrophoresis for human polyomavirus JC sequence analysis.
Five genotypes of human polyomavirus JC (JCV) have been detected so far by nucleotide sequencing and by restriction fragment length polymorphism analysis. These genotypes are the result of geographically based virus evolution. However, interesting aspects of genotyping could involve both pathogenetic and diagnostic aspects. The product from amplification of JCV sequences, found in urine from 12 healthy individuals from different countries, have been analysed by denaturing gradient gel electrophoresis (DGGE) and by nucleotide sequencing. The aim of this study was to assess if the DGGE analysis could be used to study the variability of the JCV genome. The target sequence of this study was 233 bp long, within the gene coding for the VP1, known to contain several type-determining sites. Four DGGE patterns have been observed among our strains. Five strains, from African individuals, were of type 3 and exhibited the same electrophoretic pattern, clearly distinguishable from that of the type 1 strains detected in the urine from 6 European individuals. Five type 1 strains shared a similar DGGE pattern, slightly different from that of the sixth. A different pattern characterised as a type 2 strain was detected in the urine from a Peruvian individual. These results suggest that DGGE analysis could be used as a screening assay for choosing strains for nucleotide sequencing. The analysis of a fragment larger than the one used in this study could allow the identification of more types and subtypes.
De Santis
R
R
Department of Public Health, Epidemiology and Environmental Chemistry, University of Florence, Firenze, Italy.
Azzi
A
A
D000293:Adolescent; D000328:Adult; D000368:Aged; D001739:BK Virus / Q000235:genetics; D001483:Base Sequence; D002213:Capsid / Q000235:genetics / Q000652:urine*; D036022:Capsid Proteins; D004279:DNA, Viral / Q000032:analysis / Q000652:urine*; D004586:Electrophoresis; D005260:Female; D006801:Humans; D007577:JC Virus / Q000235:genetics; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D011120:Polyomavirus / Q000235:genetics*; D017422:Sequence Analysis, DNA
D036022:Capsid Proteins; D004279:DNA, Viral; C062481:VP1 protein, polyomavirus