Nonradioactive in situ hybridization in progressive multifocal leukoencephalopathy
1Department of Neurology, Mayo Clinic, Rochester, MN 55905.
Insights
Biotinylated DNA:DNA in situ hybridization effectively detects JC virus in brain tissue, aiding diagnosis of progressive multifocal leukoencephalopathy (PML). This method correlates viral presence with cellular pathology in patients, including those with AIDS.
Area of Science:
- Neurovirology
- Molecular Pathology
- Diagnostic Techniques
Background:
- Progressive multifocal leukoencephalopathy (PML) is a demyelinating disease often associated with impaired cell-mediated immunity.
- JC virus (JCV) is the causative agent of PML, and its detection in brain tissue is crucial for diagnosis.
- Traditional diagnostic methods may have limitations in sensitivity or specificity.
Purpose of the Study:
- To evaluate the utility of biotinylated DNA:DNA in situ hybridization for detecting JC virus in brain tissues of patients with suspected PML.
- To correlate the presence of JCV with pathological changes at the cellular level.
- To assess the applicability of the technique to archival and biopsy specimens.
Main Methods:
- Biotinylated DNA:DNA in situ hybridization was performed on brain tissues from 67 patients with suspected PML.
- The technique utilized biotinylated probes to detect JC virus DNA.
- Cellular pathology, including changes in oligodendrocytes and astrocytes, was examined in conjunction with viral detection.
Main Results:
- The in situ hybridization method demonstrated specificity and ease of interpretation.
- JC virus DNA was detected in brain tissues, with an average of 1,000 copies per nucleus in infected cells.
- The technique successfully identified infected cells even in small biopsy specimens and correlated viral presence with pathological changes.
Conclusions:
- Biotinylated DNA:DNA in situ hybridization is a sensitive and specific method for diagnosing PML by detecting JC virus in brain tissue.
- The technique allows for single-cell level correlation of viral presence with neuropathology.
- This method is valuable for evaluating both archival and fresh biopsy samples, serving as a useful adjunctive diagnostic procedure.
Abstract:
Biotinylated DNA:DNA in situ hybridization was used for determining the presence of JC virus in brain tissues of 67 patients thought to have progressive multifocal leukoencephalopathy. Sixty patients had acquired immunodeficiency syndrome (AIDS) or other illnesses that decreased the cell-mediated immunity. Two patients had no underlying disease, and five others had chronic illnesses not typically associated with reduced cell-mediated immunity. In situ hybridization with biotinylated probe provides specificity and ease of interpretation. The presence of virus can be correlated at the single-cell level with attendant pathologic changes in oligodendrocytes and astrocytes. Not only archival tissue but also tiny fragments of brain biopsy material can be evaluated successfully. Quantifying the technique suggests that the nucleus of a cell labeling for JC virus DNA averages 1,000 copies of replicating genome. Identification of an infected cell is pathologically significant even when only a few such cells are present in a biopsy specimen. Biotinylated DNA or RNA probes are equally effective in identifying infected cells. In situ hybridization will likely continue to be a useful adjunctive procedure for the evaluation of brain tissue from patients suspected of having progressive multifocal leukoencephalopathy.
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