Detection of measles virus genomic sequences in sspe brain tissue by the polymerase chain reaction
- 1 April 1990
- journal article
- research article
- Published by Wiley in Journal of Medical Virology
- Vol. 30 (4), 237-244
- https://doi.org/10.1002/jmv.1890300402
Abstract
The polymerase chain reaction (PCR) was modified to detect RNA genomic sequences by generating cDNA copies of those sequences as a preliminary step. Oligonucleotide primer pairs complementary to sequences in each of the five major structural protein genes of the measles virus (nucleocapsid protein, phosphoprotein, matrix protein, fusion protein, and hemagglutinin protein) were synthesized. PCR products were tentatively identified by visualization of bands of the appropriate size by ethidium bromide staining after gel electrophoresis, and identity was confirmed by subsequent restriction enzyme cleavage of the products at predetermined sites to yield fragments of predicted size. This method successfully amplified 400–500 base regions from each of these five genes in RNA extracts of wild measles virus cultured in Vero cells and in RNA extracted from most of the SSPE brain tissues tested, but not in RNA from any control brain tissues. Measles virus genome was detected in SSPE brain tissues stored frozen for as long as 27 years and formalin-fixed paraffin-embedded subacute sclerosing panencephalitis (SSPE) brain tissues as old as 9 years. This method provides a simple, rapid and highly sensitive means of detecting and identifying sequences of RNA genomes by PCR. The success of this method in detecting measles virus in SSPE brain tissues suggest that PCR is appropriate to investigate the possible presence of RNA viruses in other neurological disorders of unknown etiology.Keywords
This publication has 27 references indexed in Scilit:
- Avoiding false positives with PCRNature, 1989
- Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA PolymeraseScience, 1988
- Fusion Glycoprotein of Measles Virus: Nucleotide Sequence of the Gene and Comparison with Other ParamyxovirusesJournal of General Virology, 1987
- Detection of Measles Virus RNA in Lymphocytes from Peripheral-Blood and Brain Perivascular Infiltrates of Patients with Subacute Sclerosing PanencephalitisNew England Journal of Medicine, 1985
- Persistent Measles Infection of the Central Nervous System: Subacute Sclerosing PanencephalitisClinical Infectious Diseases, 1983
- Isolation of biologically active ribonucleic acid from sources enriched in ribonucleaseBiochemistry, 1979
- Integration of viral genomesNature, 1975
- Subacute Sclerosing Panencephalitis: Propagation of Measles Virus from Brain Biopsy in Tissue CultureScience, 1969
- Subacute Sclerosing Panencephalitis: Isolation of Measles Virus from a Brain BiopsyNature, 1969
- MEASLES-VIRUS ANTIBODY AND ANTIGEN IN SUBACUTE SCLEROSING PANENCEPHALITISThe Lancet, 1967