【摘要】： 嗜酸性粒細(xì)胞陽離子蛋白(ECP)是一組由不同種類組成的堿性蛋白質(zhì),被廣泛應(yīng)用于過敏性患者臨床檢驗(yàn)的標(biāo)志分子,也是酸性粒細(xì)胞活性和更新的一個指標(biāo)。 本實(shí)驗(yàn)以ECP為抗原,制備了多克隆抗體和單克隆抗體,經(jīng)辛酸-硫酸銨和蛋白親和層析及濃縮后,多克隆抗體和單克隆抗體效價均達(dá)到1:10~7。初步建立了基于ECP的雙抗夾心ELISA方法,其檢測范圍為10ng/ml-900ng/ml,檢測下限為5ng/ml。 將方法應(yīng)用于戊型肝炎患者血清樣本進(jìn)行檢測,其中:戊型抗原Ag陽性樣本9個,IgG陽性樣本100個,IgM陽性樣本72個,普通健康人群血清樣本10個。檢測結(jié)果:戊肝陰性樣本ECP水平最高,其次為戊肝IgG陽性樣本,Ag陽性樣本ECP水平較低,IgM陽性樣品ECP水平最低。t檢驗(yàn)結(jié)果顯示:IgM、Ag陽性樣本兩者差異顯著,有統(tǒng)計學(xué)意義(P＜0.005),而戊肝IgG陽性樣本與戊肝陰性樣本無統(tǒng)計學(xué)意義(0.10＜P＜0.25),戊型肝炎病毒可能抑制了嗜酸性粒細(xì)胞的活性,因而戊型肝炎早期陽性血清樣本中ECP的含量較低,感染后期接近健康水平陽性血清樣本中ECP的含量接近陰性血清,該實(shí)驗(yàn)結(jié)果國內(nèi)首次報道。 本結(jié)果為進(jìn)一步研究嗜酸性粒細(xì)胞ECP的釋放水平與戊型肝炎病毒感染的相關(guān)性與及做為戊型肝炎的一種早期診斷標(biāo)志物提供了實(shí)驗(yàn)基礎(chǔ)。
[Abstract]:Eosinophil cationic protein (ECP) is a group of basic proteins composed of different species. It is widely used as a marker for clinical examination in allergic patients and is also an indicator of activity and renewal of acidophilic granulocytes. In this study, polyclonal antibodies and monoclonal antibodies were prepared using ECP as antigen. The titers of polyclonal antibodies and monoclonal antibodies reached 1: 10107after affinity chromatography and concentration of caprylic acid-ammonium sulfate and protein. A double antibody sandwich ELISA method based on ECP is established. The detection range is 10 ng / ml-900ng / ml and the detection limit is 5 ng / ml. The method was applied to the detection of hepatitis E patients' serum samples, including 9 positive samples of Ag, 100 positive samples of IgG, 72 positive samples of IgM, and 10 samples of serum from healthy people. The results showed that ECP level was the highest in hepatitis E negative samples, followed by IgG positive samples in hepatitis E, ECP level in Ag positive samples was lower, ECP level in IgM positive samples was the lowest. T test results showed that IgM,Ag positive samples had significant difference between them. There was statistical significance (P < 0.005), but there was no significant difference between IgG positive samples and negative samples (0.10 < P < 0.005). Hepatitis E virus might inhibit the activity of eosinophils. As a result, the content of ECP in the early positive serum samples of hepatitis E was lower than that in the positive serum samples at the later stage of infection, and the content of ECP in the positive serum samples was close to the negative serum. The results of the experiment were reported for the first time in China. The results provide experimental basis for further study on the correlation between eosinophil ECP release level and hepatitis E virus infection and as an early diagnostic marker for hepatitis E.
【學(xué)位授予單位】：長春理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R392

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