Detection of TGF-?RⅡ mRNA in peripheral blood mononuclear cells from rheumatoid arthritis patients using nested-real-time fluorescence quantitative PCR
- VernacularTitle:巢式实时荧光定量PCR检测RA患者外周血单个核细胞TGF-?RⅡmRNA
- Author:
Chunyan LIU
;
Yun WANG
;
Liwen WANG
- Publication Type:Journal Article
- Keywords:
TGF-? RLL;
rheumatoid arthritis;
nested-real-time fluorescence quantitative PCR;
gene expression
- From:
Chinese Journal of Clinical Laboratory Science
2006;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop a nested-real-time fluorescence quantitative PCR method for detection of TGF-?RII mRNA in human peripheral blood mononuclear cells(PBMC),and analyze the clinical implications of TGF-? RII in RA patients.Method The outer primers of span intron and inner primers were designed for nested-real-time fluorescence quantitative PCR.To increase specific template,the concentration of primers and enzyme,cycle times in the first amplification were decreased,and the real-time fluorescent quantity assay was performed in the second amplification.The ?-actin gene was used as internal reference,and the results were presented as the ratios of TGF-? RII mRNA to ?-actin mRNA.Results The standard curve showed a fine linear relationship between Ct(cycle threshold)and logarithm of template concentration,and the correlation coefficient was 0.999.The sensitivity reached 101 copies.The TGF-? RII gene expression in PBMC of RA patient was increased compared to healthy controls[(0.45?0.11)vs(0.37?0.10),P
