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Published on: February 29, 2020
No evidence of measles virus in stapes samples from patients with otosclerosis
A B Grayeli1, P Palmer, P Tran Ba Huy
1INSERM U.426, Faculté Xavier Bichat, Université Paris 7, France. grayeli@bichat.inserm.fr
Abstract:
Otosclerosis is a localized bone dystrophy of unknown etiology mainly involving the stapes. The hypothesis of a persistent infection by the measles virus was based on the inconstant detection of the virus by various methods, including reverse transcription-PCR (RT-PCR) of patients' stapes samples. The aim of this work was to investigate the presence of the measles virus in stapedial otosclerosis foci by different sensitive methods. Pathologic stapes samples were obtained from 35 patients suffering from otosclerosis. Measles virus detection was performed by (i) cocultures of Vero cells and primary cell cultures of bone samples (n = 7), (ii) immunofluorescence study of these cocultures (n = 3), and (iii) RT-PCR on RNA directly obtained from fresh frozen samples (n = 28) and on RNA extracted from the primary cell cultures (n = 2). Viral genomic regions coding for N (nucleoprotein) and M (matrix) proteins were separately amplified. PCR sensitivity was optimized on the measles virus Edmonston strain. Glyceraldehyde-3-phosphate dehydrogenase mRNA was used as a marker of total RNA recovery. PCR products were tested by Southern blot hybridization technique to improve sensitivity and specificity. PCRs amplifying the M and the N protein genes were able to detect the control measles virus RNA at titers as low as 0.1 and 0.01 50% tissue culture infective dose, respectively. With these highly sensitive methods, we could not evidence the presence of the measles virus in any of our bone samples or primary bone cell cultures. Our results do not confirm the hypothesis of persistent measles virus infection in otosclerosis.
Insights
This study investigated measles virus in otosclerosis bone samples using sensitive methods. Researchers found no evidence of measles virus, refuting the persistent infection hypothesis in otosclerosis patients.
Area of Science:
- Otolaryngology
- Virology
- Molecular Biology
Background:
- Otosclerosis is a bone disorder affecting the stapes, with an unknown cause.
- A persistent measles virus infection has been hypothesized as a potential cause of otosclerosis.
- Previous detection methods for measles virus in otosclerosis samples have yielded inconsistent results.
Purpose of the Study:
- To investigate the presence of measles virus in stapedial otosclerosis foci using highly sensitive detection methods.
- To determine if measles virus plays a role in the etiology of otosclerosis.
Main Methods:
- Pathologic stapes samples from 35 otosclerosis patients were analyzed.
- Methods included cell cocultures, immunofluorescence, and highly sensitive reverse transcription-polymerase chain reaction (RT-PCR).
- RT-PCR targeted viral nucleoprotein (N) and matrix (M) genes, with results confirmed by Southern blot hybridization.
Main Results:
- The optimized RT-PCR methods demonstrated high sensitivity, detecting control measles virus RNA at low titers.
- Despite employing sensitive techniques, the measles virus was not detected in any of the bone samples or primary bone cell cultures.
- No evidence of measles virus was found in the stapedial otosclerosis foci examined.
Conclusions:
- The findings do not support the hypothesis of a persistent measles virus infection in otosclerosis.
- The etiology of otosclerosis remains unknown, and measles virus is unlikely to be a contributing factor based on this study.
- Further research may be needed to explore other potential causes of this localized bone dystrophy.
