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Detection of virus genomic sequence in perilymph of deaf patient by using the polymerase chain reaction

Research Project

Project/Area Number 10671592
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Otorhinolaryngology
Research InstitutionOsaka 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
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1999: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1998: ¥1,800,000 (Direct Cost: ¥1,800,000)
Keywordsvirus infection of the inner ear / measles virus / RT-PCR / Deafness / ウイルス / 内耳炎
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.

Report

(3 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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