Detection of JC Virus DNA by Polymerase Chain Reaction in Patients with Progressive Multifocal Leukoencephalopathy

Abstract
Polymerase chain reaction (PCR) was used in the detection of JC virus (JCV) DNA in formalinfixed, paraffin-embedded brain tissue sections from 24 patients with progressive multifocalleukoencephalopathy. Brain sections from normal autopsies (n = 17)and other neurologic conditions (n = 4) were used as controls. Specific amplified products were detected in 20 (83%) of 24 patients with progressive multifocalleukoencephalopathy. PCR did not amplify JCV or human β-globingene sequences in four patients with characteristic demyelinating lesions of progressive multifocalleukoencephalopathy that were positive for JCV by in situ hybridization or immunocytochemistry. PCR from biopsy sections resulted in more intense amplification signals than PCR from autopsy tissue. Normal brain sections from 17autopsies were negative by PCR. Low-grade amplification of JCV was observed in one patient with herpes simplex virus encephalitis. PCR served as a more rapid test for confirmation of progressive multifocalleukoencephalopathy than in situ hybridization. However, PCR performance was altered by prolonged formalin fixation of tissue and undetermined inhibitors of the amplification reaction. Laboratories and clinicians should be aware of the difficulties encountered when using paraffin-embedded tissue for PCR.