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Diagnosis of progressive multifocal leucoencephalopathy by hybridisation techniques
R H Boerman1, E P Arnoldus, A K Raap
1Department of Neurology, University Hospital, Leiden, The Netherlands.
Abstract:
In situ hybridisation with acetyl-aminofluorene (AAF) and 35S-labelled DNA probes for polyomaviruses, was used to detect JC virus DNA in brain necropsy material in a patient with progressive multifocal leucoencephalopathy (PML). In a second patient PML was diagnosed from a brain necropsy specimen using the same technique. The main infected cell type were oligodendrocytes; dot hybridisation was used to estimate the number of viral copies in each infected cell. Southern blot hybridisation for further analysis of the viral genome was also carried out. In situ hybridisation with non-radioactive labelled polyomavirus DNA provides a simple and specific means for studying viral DNA in formaldehyde fixed tissue sections from patients with suspected PML. Even in small biopsy samples hybridisation results can be correlated with standard histopathological, immunocytochemical, and electron microscopic findings.
Insights
In situ hybridisation detected JC virus DNA in brain tissue from patients with progressive multifocal leucoencephalopathy (PML). This method identifies viral DNA in infected oligodendrocytes, aiding PML diagnosis.
Area of Science:
- Virology
- Neuroscience
- Molecular Biology
Background:
- Progressive multifocal leucoencephalopathy (PML) is a demyelinating disease of the central nervous system.
- JC virus (JCV), a polyomavirus, is the causative agent of PML.
- Accurate and timely diagnosis of PML is crucial for patient management.
Observation:
- In situ hybridisation (ISH) using acetyl-aminofluorene (AAF) and 35S-labelled DNA probes for polyomaviruses was employed.
- JC virus DNA was successfully detected in brain necropsy material from two PML patients.
- The primary infected cell type identified was oligodendrocytes.
Findings:
- ISH enabled the detection of JC virus DNA in formaldehyde-fixed brain tissue sections.
- Dot hybridisation quantified viral copies within infected cells.
- Southern blot hybridisation allowed for further analysis of the viral genome.
Implications:
- Non-radioactive labelled polyomavirus DNA ISH offers a simple, specific method for studying viral DNA in suspected PML cases.
- This technique can be applied to small biopsy samples.
- ISH findings correlate well with standard histopathological, immunocytochemical, and electron microscopic diagnostic methods.