C. Sugimoto et al., AMPLIFICATION OF JC VIRUS REGULATORY DNA-SEQUENCES FROM CEREBROSPINAL-FLUID - DIAGNOSTIC-VALUE FOR PROGRESSIVE MULTIFOCAL LEUKOENCEPHALOPATHY, Archives of virology, 143(2), 1998, pp. 249-262
Progressive multifocal leukoencephalopathy (PML) is a fatal demyelinat
ing disease in the central nervous system caused by a ubiquitous human
polyomavirus designated as JC virus (JCV). PML affects individuals wi
th decreased immune competence and is now one of the common opportunis
tic infections in patients with AIDS. JCV DNAs in the brain of PML pat
ients contain various PML-type regulatory regions that were generated
from the archetypal regulatory region during persistence. Recently, ma
ny studies have suggested that detection of JCV DNA from the cerebrosp
inal fluid (CSF) may offer a tool for diagnosing PML. However, in all
of these studies, coding Sequences within the T antigen or capsid prot
ein gene have been targeted for amplification. To amplify the JCV regu
latory region, we established a nested PCR that could efficiently ampl
ify the regulatory region from most JCV subtypes prevalent in the worl
d. Using this PCR, we amplified JCV regulatory regions from the CSF sa
mples from 4 patients strongly suspected of PML, whereas amplification
was negative from 80 CSF samples from patients without PML. Sequencin
g of the amplified fragments PML patients, we analyzed the structures
of regulatory regions derived from the brain as well as CSF. In each o
f these cases, the major regulatory sequence of both origins were iden
tical. This finding indicates that JCV DNA in brain lesions is excrete
d in the CSF. Since the structures of PML-type JCV regulatory regions
are unique to individual patients, the current PCR, if the amplified f
ragments are sequenced, can eliminate false positives that may arise f
rom contaminations.