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Unit Code 81107:
JC Virus (JCV) Detection by In Situ Hybridization

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Method Description

The probes are labeled with digoxigenin and used in an in situ

hybridization procedure with hybridization for 2 hours. After

washing, the tissues are incubated with anti-digoxigenin antibody

conjugated to alkaline phosphatase and the hybridization product

is detected by reaction of the enzyme conjugate with the substrate

5-bromo-4-chloro-3-indolyl phosphate (BCIP) and concomitant

reduction of nitroblue tetrazolium (NBT). Some bizarre, malignant-

appearing astrocytes may stain with the JCV probe. These cells are

infected by JCV in PML.  Morphologically, these cells have enlarged

nuclei (25-60 nm), lobulated nuclei, multiple nuclei, nuclear

hyperchromasia with a coarse chromatin pattern, and occasional frank

mitosis. (Aksamit AJ, Mourrain P, Sever JL, Major EO:  Progressive

multifocal leukoencephalopathy: investigation of three cases using in

situ hybridization with JC virus biotinylated DNA probe. Ann Neurol

1985;18:490-496)

Performing Laboratory Location

Rochester

Key