1999 Fiscal Year Final Research Report Summary
Detection of virus genomic sequence in perilymph of deaf patient by using the polymerase chain reaction
| Project/Area Number |
10671592
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Otorhinolaryngology
|
|---|
| Research Institution | Osaka University Graduate School of Medicine |
|---|
Principal Investigator |
TAMURA Manabu Osaka University, Graduate School of Medicine, Assistant Professor, 医学系研究科, 助教授 (50273644)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
DOI Katumi Osaka University, Graduate School of Medicine, Assistant Professor, 医学系研究科, 助教授 (40243224)
|
|---|
| Project Period (FY) |
1998 – 1999
|
| Keywords | virus infection of the inner ear / measles virus / RT-PCR / Deafness |
|---|
| Research Abstract |
The etiology of the profound sensory hearing loss is mostly unknown. Virus infection of the inner ear infection is one of the important cause of acquired deafness. The aim of our study was to detect measles virus genome in the perilymph of the affected patients using RT-PCR system. Ten μ l of the perilymph was obtained during the surgery of cochlear implant and frozen until the performance of RT-PCR.Total RNA was extracted by the guanidinium thiocyanate-phenol-chloroform method. The extracted RNA was submitted to reverse transcription (RT) With random primers. The cDNA was then amplified by nested PCR using primers specific for nucleoprotein (NP) of measles virus. Identity of PCR product was confirmed by direct sequence method. NP gene of measles virus was detected four out of 5 samples by RT-PCR.Seven nucleotide substitutions were present in 4 samples compared with Edomonston strain and CAM strain. Deduced amino acid sequences showed 2 mutations in 4 samples compared with Edomonston strain and CAM strain. The perilymph obtaqined from the patients of otosclerosis was also analyzed by this RT-PCR method. Three nucleotide substitutions were present in 3 samples compared with Edomonston strain and CAM strain. Deduced amino acid sequences showed 1 mutations in 3 samples compared with Edomonston strain and CAM strain.
|
