| Project/Area Number |
07670881
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Pediatrics
|
|---|
| Research Institution | Sapporo Medical University School of Medicine |
|---|
Principal Investigator |
NAKATA Shuji Sapporo Medical University, School of Medicine, Department of Pediatrics, Assistant Professor, 医学部, 講師 (70155745)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
KOGAWA Keiko Sapporo Medical University, School of Medicine, Department of Pediatrics, Instru, 医学部, 助手
ADACHI Noriaki Sapporo Medical University, School of Medicine, Department of Pediatrics, Instru, 医学部, 助手
|
|---|
| Project Period (FY) |
1995 – 1996
|
| Project Status |
Completed (Fiscal Year 1996)
|
| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1996: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1995: ¥1,300,000 (Direct Cost: ¥1,300,000)
|
|---|
| Keywords | HuCV / Sa / 82 / J / Infantile gastroenteritis / ELISA / RDRP region / RT-PCR / Sequence analys / Dot Blot Hybridization / Human calicivirus |
|---|
| Research Abstract |
Besed on genome analysis of the RNA dependent RNA polymerase (RDRP) region, it has been proposed that human calicivirus (HuCV) can be classified into at least three genogroups : genogroup I is represented by Norwalk virus (NV), genogroup II by Snow Mountain virus (SMV) and genogroup III by HuCV/Sapporo/82/Japan (HuCV/Sa/82/J) virus. Among three genogroups, the sequnece homology in RDRP region is less than 70% and antigenic relatedness has not been demonstrated. The RDRP region of HuCV/Sa/82/J-related strains collected from children in Sapporo between 1979 and 1990 was amplified by RT-PCR and sequenced. Nucleotide and amino acid sequences of the PCR products showed a high degree of identity among above samples and prototype of HuCV/Sa/82/J.These data indicate a correlation between the antigenicity and sequence similarity of the RDRP region among HuCV/Sa/82/J-related viruses. This virus has been circulating in Sapporo for at least 10 years. These strains detected in USA,UK and Saudi Arabia also showed a high degree of identity in the RDRP region to prototype of virus.
A dot blot hybridization assay with a cDNA probe derived from the RDRP region of HuCV/Sa/82/J was developed for detection of HuCv/Sa/82/J.This assay was specific for HuCV/Sa/82/J and related viruses, and the sensitivity was about 10^5 physical particles or 10pg of cDNA.The entire genome of NV cDNA and feline calicivirus RNA did not hybridize under high stringent conditions of stringency with the HuCV/Sa/82/J cDNA.A higher positive rate for virus detection in stool samples was obtained with the dot blot assay (21%) than ELISA (10%). The dot blot assay is specific, easy to perform and advantageous with unlimited supply of reagents, and should be useful for epidemiological and molecular biological studies of HuCV/Sa/82/J and related strains.
|
