The detection of the Nucleic Acid of Human Coronaviruses 229E and OC43 in Tissue Specimens Using the Reverse Transcription-Polymerase Chain Reaction

Stewart, Janet (1992). The detection of the Nucleic Acid of Human Coronaviruses 229E and OC43 in Tissue Specimens Using the Reverse Transcription-Polymerase Chain Reaction Mémoire. Québec, Université du Québec, Institut National de la Recherche Scientifique, Maîtrise en virologie et immunologie, 173 p.

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Résumé

On the basis of epidemiologie studies, an environmental agent, particularly a virus, is suspected of being the cause of multiple sclerosis. Human coronaviruses number among the potential etiologie agents due to several observations. Coronavirus-like particles were observed by the electron microscope in a brain section from a patient with multiple sclerosis and antibody to human coronaviruses 229E and OC43 are detectable in the cerebrospinal fluid of a significant proportion of MS patients. The present project involved the setting up of the reverse transcription-polymerase chain reaction technique for the detection of human coronaviruses, HCV-229E and HCV-OC43, and a preliminary study of human tissue specimens, including CNS tissues from multiple sclerosis patients, for the presence of the nucleic acid of the two viruses. The quality of the RNA extracted from archival tissues was verified using an additional RT-PCR test to detect cellular messenger RNAs in them, either human MBP RNA in white matter or r-actin mRNA in PBLs and in grey matter. HCV-229E RNA could be detected in sorne specimens and also the limiting number of targets detectable by this RT-PCR method was estimated at 60,000. Peripheral blood lymphocytes from 13 healthy adults were analyzed for the presence of nucleic acid from beth viruses and three specimens of the 13 were positive in two consecutive tests for HCV-229E RNA. While this is a small number of specimens, this result is very promising in terms of future characterization of the tropism of HCV-229E. Failure to detect HCV-OC43 RNA indicates that these cells are not susceptible to infection by this virus, however, as in the case of HCV-229E, a larger number of individuals should be tested in arder to confirm these observations. A study of CNS tissue specimens from multiple sclerosis patients and controls was also conducted. Four of 21 specimens from 11 MS patients were positive when tested for HCV-229E RNA and were confirmed positive in subsequent tests. None of 11 specimens from the control brains tested positive nor did any of the CNS tissues show evidence for the presence of HCV-OC43 RNA. Our results in CNS tissues emphasize the importance of continuing studies on the possible implication of HCV-229E in neurological diseases.

Type de document:	Thèse Mémoire
Directeur de mémoire/thèse:	Talbot, Pierre
Mots-clés libres:	-
Centre:	Centre INRS-Institut Armand Frappier
Date de dépôt:	08 août 2019 01:50
Dernière modification:	23 mai 2023 14:06
URI:	https://espace.inrs.ca/id/eprint/8527

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