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중합효소연쇄 반응을 이용한 신생아 소변에서 Cytomegalovirus측정 및 방법
최철석,김은아,이경옥,이규범,이연태 대한바이러스학회 1994 Journal of Bacteriology and Virology Vol.24 No.1
Polymerase Chain Reaction(PCR) was used to detect human cytomegalovirus(HCMV) in urine sample of an infant with neonatal hepatitis. The patient was HCMV seropositive for IgG, IgM by ELISA, and also showed high level of aminotransferase(500I.U.) and direct bilirubine ( >10mg/dl). Several reports have described difficulties of PCR in urine samples. This report is a rapid and reliable detection method of HCMV DNA in urine from a patient by extraction with guanidinium thiocyanate/silica. This method is based on lysing and adsorption of DNA for PCR amplification. Urine sample prepared by this method should good result. In our opinion, PCR with this method is sensitive, rapid and reliable procedure for successful PCR for detection of HCMV in urine sample.
임상 혈청에서 중합효소 연쇄반응을 이용한 HCV RNA 측정
최철석,김은아,박영석,이경옥,이규범,이연태 대한바이러스학회 1994 Journal of Bacteriology and Virology Vol.24 No.1
In 152 sera obtained from probable patients with hepatitis seroprevalence of antibody against hepatitis C virus(HCV) by ELISA and the viral genome by reverse transcriptase polymerase chain reaction (RT-PCR) were detected. Anti-HCV positive rate by enzyme-linked immunosor- bent assay(ELISA) were 23.0% (35/152), hepatitis C virus RNA positive were 40.7% (62/152). Of 152, 25(16.4% ) were positive either anti-HCV or RT-PCR. Ten cases(6.5 ) were anti- HCV positive whereas RT-PCR was shown negative. While, 37 cases(24.3 % ) were negative for anti-HCV with positive result for RT-PCR. Eighty sera of 152 were negat.ive by both methods. In this study the PCR method provided higher sensitivity than ELISA method for detection of HCV. It is our concluded that the presence of hepatitis C viral genome detected by RT-PCR in sera is a better test of infectivity than the anti-HCV method.
질내가검물 및 소변에서 중합효소연쇄반응을 이용한 Chlamydia trachomatis 검사
최철석 ( Cheol Seok Choi ),성상철 ( Sang Chul Seong ),이무주 ( Mu Ju Lee ) 대한임상검사과학회 1993 대한임상검사과학회지(KJCLS) Vol.25 No.1
A practical test using polymerase chain reac-tion(PCR) for detection of Chlamyadia tracho-matis in clinical samples was performed. Also thets for specificity and sensitivity of proim-ers for 537 base pair fragment were deter-mined. In this study, 20 specimens (15 viginal swabs, 5 urines of men) with non-gonocooccal urethritis (NGU) were subjected to the PCR test. For specificity of test, C. trachomatis serovar (ATCC) for positive control, C. psittaci, N. gonorrheae for negative control were used, respectively. For sensitlvity test, extracted C. trachomatis DNA (36ng/㎕) was serial 10-fold diluted to 10-8. DNA was extracted with phenol/chloroform after proteinase K treatment and precipitated with ethanol. The PCR products were analysed by electrophoresis on 2.0% agarose gels and the DNA was stained with ethidium bromide. Seven (35.0%) of 20 samples tested were PCR positive for C. trachomatis. Of these positive specimens, 5 were viginal swabs, 2 were urine. In specificity of detection, C. trachomatis DNA was amplified with 537 base pair DNA fragment, but all negative control strain DNAs were not amplified. The sensitivity of PCR detection with primers used was as-sessed at 2.6fg of input DNA. In this study the PCR provided sensitive and specific tool for detection f Chlamydia tracho-matis. Accordingly it is our opinon that PCR technique with proper primers is rapid and re- liable tool for the diagnosis of Chlamydia trachomatis infection.