| Project/Area Number |
03670750
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Urology
|
|---|
| Research Institution | Gifu University |
|---|
Principal Investigator |
BAN Yoshihito Gifu University School Associate Professor of Medicine, 医学部, 助教授 (30021497)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
DEGUCHI Takashi Gifu University School Assistant Professor of Medicine, 医学部・附属病院, 講師 (40163935)
伊藤 康久 岐阜大学, 医学部, 助手 (50223194)
|
|---|
| Project Period (FY) |
1991 – 1993
|
| Project Status |
Completed (Fiscal Year 1993)
|
| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1993: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1992: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1991: ¥800,000 (Direct Cost: ¥800,000)
|
|---|
| Keywords | Non-gonococcal urethritis / U.urealyticum / PCR / DNA診断 / Ureaplasma urealyticum / 尿道炎 |
|---|
| Research Abstract |
A DNA fragment of 177 bp was amplified by the Polymalase chain reaction(PCR) procedure, with use of a set of oligonucleotides based on sequences within the 16S ribosomal RNA gene from Ureaplasma urealyticum as an extention primer for the PCR.No amplified production was detected from the bacterial DNA including those of Chlamydia trachomatis, Neisseira gonorhoeae, Mycoplasma hominis, Mycoplasma pneumoniae and gram- positive and/or gram- negative uropathogens such as Escherichia coli etc. The amplified DNA fragment of 177 bp was detected on agarose gel electrophoresis, when DNA of 4.4X10^2 cells of U.urealyticum per PCR was used as template for the PCR.This method for the detection of U.urealyticum was very specific and sensitive, and all the procedures can be done quickly within 6 hours.
Because of the need for a painless specimen sampling such as first-voided urine sampling, we tried detection of U.urealyticum by the PCR using first-voided urine as a specimen, and the results was compared with the results obtained by the ordinary T-broth method. Fourty-six male urethritis patients were tested by both methods, resulting a positive coincidence rate of 68%(15/22) and negative coincidence rate of 100%(24/24). The reason of the low detection rate by PCR than the detection rate by ordinary T-broth method was thought that false-negative results of PCR was possibly due to some PCR inhibitting factor in the urine and false-positive results of PCR was possibly due to bacteria other than U.urealyticum.
Therefore, we improved the method culturing the T-broth medium which changed to red color and/or changing the DNA preparation method from urine to prevent the inhibition by some PCR inhibitting factor in the urine.
|
