摘要
目的：研究血吸虫病肝纤维化发病机理、确定肝纤维化活动程度、判断治疗效果和预后等，建立定量ＰＣＲ法。方法：从引物合成构建质粒始，建立ＲＴ－ＰＣＲ加ｄｏｔｂｌｏｔ法测ＰＢＭＣ中ＴＧＦ－β１ｍＲＮＡ水平，以构建的ＴＧＦ－β１质粒，精确定量后作标准对照，以看家基因β－ａｃｔｉｎ同管进行ＲＴ－ＰＣＲ为内对照，ｄｏｔｂｌｏｔ结果以多功能凝胶成像仪ＩＳ－１０００定量分析，检测梯度稀释的质粒和１１份不同血标本的复管。结果：８个梯度稀释的ＴＧＦ－β１质粒ＰＣＲ产物定量结果与原浓度对数成正比，１１份标本两次结果为１．７１±０．９０和１．５４±０．８８，显示较好的重复性和稳定性。结论：该方法可用于定量分析ＰＢＭＣ中ＴＧＦ－β１ｍＲＮＡ含量。
AIM: To establish a method to measure the TGF β 1 mRNA level for studying the mechanism of fibrogenesis caused by schistosomiasis japonica.METHODS: Reverse transcription polymerase chain reaction and dot blot analysis were used. A plasmid of TGF β 1 was constructed for standardization,and β actin was used as control. Eight different concentrations of the plasmid and 11 double tube of TGF β 1 mRNA in the peripheral blood mononuclear cell (PBMC) of different persons were measured. RESULTS: Quantitative results of 8 different dilutions of TGF β 1 plasmid had positive with the logarithm of the original concentration. The results of the 11 double tube were 1.71±0.90 and 1.54±0.88.CONCLUSION: The duplicability and stability of the method showed it can be used to analyse the TGF β 1 mRNA level of the peripheral blood mononuclear cells.
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
1998年第3期219-222,共4页
Chinese Journal of Parasitology and Parasitic Diseases
基金
总理基金
