title: Co-infection with two JC virus genotypes in brain, cerebrospinal fluid or urinary tract detected by direct cycle sequencing of PCR products.
abstract: The human polyomavirus JC (JCV), which exists in different geographically based genotypes, causes the central demyelinating disease known as progressive multifocal leukoencephalopathy (PML). A coding region recombinant JCV Type 1/Type 3 (Type 4) is excreted in the urine of some 16% of individuals in the USA. In addition, occasional 'crossovers' in viral DNA sequence at type-specific sites in the coding region occur between JCV genotypes amplified from PML brain. For recombination to occur requires the existence of two different genotypes in the same host. Here we provide evidence from direct cycle sequencing of PCR products that different genotypes of JCV can be found in a single tissue sample. After non-type-specific PCR amplification, cycle sequencing produced 'split bands' at type determining sites which were resolved into type or subtype-specific sequences by subcloning of the PCR products. PCR products with split bands at typing sites were found in two brain samples and in one cerebrospinal fluid (CSF) from AIDS patients with PML and in the urine of four immunocompetent individuals. This indicates that co-infection with two viral types does not depend on severe immunocompromise. Combinations of genotypes found were Types 1A & 1B, 1A & 2, 1B & 2 and 2 & 3. In one doubly infected patient the major JCV type excreted in the urine changed within 1 week.
authors.affiliation: Laboratory of Experimental Neuropathology, NINDS National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: H T
authors.initials: HT
authors.lastname: Agostini
authors.forename: C F
authors.initials: CF
authors.lastname: Ryschkewitsch
authors.forename: E J
authors.initials: EJ
authors.lastname: Singer
authors.forename: G L
authors.initials: GL
authors.lastname: Stoner
mesh_terms: D000328:Adult; D001483:Base Sequence; D001921:Brain / Q000821:virology*; D002555:Cerebrospinal Fluid / Q000821:virology*; D004279:DNA, Viral / Q000032:analysis / Q000302:isolation & purification; D005260:Female; D005838:Genotype; D006801:Humans; D007577:JC Virus / Q000235:genetics / Q000302:isolation & purification*; D007968:Leukoencephalopathy, Progressive Multifocal / Q000821:virology; D008297:Male; D008969:Molecular Sequence Data; D009103:Multiple Sclerosis / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D014551:Urinary Tract / Q000821:virology*
chemical_list: D004279:DNA, Viral