基于自身紅細(xì)胞構(gòu)建的溶血系統(tǒng)檢測(cè)總補(bǔ)體活性及其與MICA基因多態(tài)性的關(guān)系
發(fā)布時(shí)間：2018-07-31 17:18
【摘要】：目的:本文在CH50溶血法的基礎(chǔ)上,以人紅細(xì)胞替代綿羊紅細(xì)胞,將多管法轉(zhuǎn)化為單管法,創(chuàng)建一種操作簡(jiǎn)單,穩(wěn)定可靠的檢測(cè)血漿補(bǔ)體總活性的新方法。同時(shí)本文應(yīng)用該法檢測(cè)ICU患者,冠心病患者,健康體檢者的血漿補(bǔ)體總活性,并在分子水平上研究了冠心病患者補(bǔ)體活性與MICA*008基因攜帶的關(guān)系。方法:1.首先以綿羊紅細(xì)胞替代人紅細(xì)胞,兔抗人紅細(xì)胞抗體替代兔抗羊紅細(xì)胞抗體(溶血素)進(jìn)行溶血試驗(yàn),證明不同紅細(xì)胞濃度和血漿濃度對(duì)紅細(xì)胞破壞后釋放血紅蛋白效能的影響。并通過(guò)CH50多管法溶血試驗(yàn)確定發(fā)生溶血反應(yīng)的各種試劑的最適濃度,創(chuàng)建單管溶血比色法。同時(shí)通過(guò)線性研究和重復(fù)試驗(yàn)驗(yàn)證了該法的準(zhǔn)確度和穩(wěn)定性。2.使用CH50標(biāo)準(zhǔn)法檢測(cè)血漿和血清中的補(bǔ)體活性,并使用單管溶血比色法檢測(cè)以不同人紅細(xì)胞作為溶血指示系統(tǒng)的同一份血漿中的補(bǔ)體活性,進(jìn)一步驗(yàn)證單管溶血比色法的可行性。3.使用CH50法和單管溶血比色法檢測(cè)同一份血漿的補(bǔ)體活性,驗(yàn)證兩種方法的相關(guān)性。4.使用單管溶血比色試驗(yàn)檢測(cè)ICU患者,冠心病患者和健康體檢者的補(bǔ)體總活性,證明該方法有一定的臨床應(yīng)用價(jià)值。同時(shí)采用PCR技術(shù)檢測(cè)冠心病患者和健康者的MICA*008基因攜帶情況,分析冠心病患者補(bǔ)體活性與MICA*008基因的關(guān)系。結(jié)果:1.本文應(yīng)用人紅細(xì)胞和兔抗人紅細(xì)胞抗體替代綿羊紅細(xì)胞和溶血素進(jìn)行溶血試驗(yàn),結(jié)果發(fā)現(xiàn)不同濃度的紅細(xì)胞對(duì)溶血程度的影響低于不同濃度的血漿對(duì)溶血程度的影響,同時(shí),本文得出當(dāng)血漿濃度為20%時(shí),紅細(xì)胞濃度的不同對(duì)紅細(xì)胞破壞后釋放血紅蛋白的效能影響最低。因此本文將20%紅細(xì)胞懸液所對(duì)應(yīng)第九管(20%血漿濃度)中各成分的含量,作為單管溶血比色法中各成分比例的參考。本文應(yīng)用單管溶血比色法檢測(cè)不同濃度血漿的補(bǔ)體活性,并連續(xù)5天測(cè)定同一份血漿的補(bǔ)體活性,結(jié)果證明不同濃度血漿的補(bǔ)體總活性呈線性分布,連續(xù)5天所測(cè)定的補(bǔ)體活性值的變異系數(shù)CV=11.3%,說(shuō)明單管溶血比色試驗(yàn)具有一定的準(zhǔn)確性和穩(wěn)定性。2.通過(guò)CH50標(biāo)準(zhǔn)法檢測(cè)血漿和血清中的補(bǔ)體活性,本文得出血漿和血清中的補(bǔ)體活性完全相同,因此可以使用血漿替代血清標(biāo)本進(jìn)行補(bǔ)體活性測(cè)定,補(bǔ)體檢測(cè)技術(shù)得到了優(yōu)化。同時(shí),通過(guò)不同人紅細(xì)胞作為溶血指示系統(tǒng)檢測(cè)同一份血漿中的補(bǔ)體活性,本文得出血漿中的補(bǔ)體活性大致相同(P0.05),說(shuō)明不同紅細(xì)胞不會(huì)影響血漿中的補(bǔ)體含量,單管溶血法具有可行性。3.該方法中測(cè)定的溶血OD值與CH50溶血法所測(cè)的補(bǔ)體活性值兩者呈線性相關(guān),說(shuō)明單管溶血比色法與CH50法具有顯著的相關(guān)關(guān)系,單管溶血比色法可以替代CH50溶血法檢測(cè)補(bǔ)體總活性。4.ICU患者和冠心病患者的補(bǔ)體活性要顯著低于健康體檢者的補(bǔ)體活性,單管溶血比色法有一定的臨床應(yīng)用價(jià)值。冠心病患者和健康體檢者的MICA*008基因分布大致相同,同時(shí)冠心病患者的補(bǔ)體水平與MICA*008的攜帶沒(méi)有顯著關(guān)聯(lián)。結(jié)論:1.單管溶血法檢測(cè)補(bǔ)體活性具有良好的準(zhǔn)確性,穩(wěn)定性和可行性,并且與CH50法具有顯著的相關(guān)關(guān)系,該法進(jìn)一步優(yōu)化CH50溶血試驗(yàn),具有一定的臨床應(yīng)用價(jià)值。2.ICU患者和冠心病患者的補(bǔ)體活性較低,說(shuō)明ICU患者的機(jī)體免疫功能紊亂,冠心病患者機(jī)體或許處于炎癥反應(yīng)階段。3.冠心病患者的補(bǔ)體水平與MICA*008基因沒(méi)有顯著關(guān)聯(lián)。
[Abstract]:Objective: on the basis of CH50 hemolysis, we use human erythrocytes to replace sheep red blood cells and convert multi tube method into single tube method to create a new method to detect the total complement activity of plasma complement, which is simple and stable and reliable. At the same time, this method is used to detect the total complement activity of ICU patients, coronary heart disease patients and health examiners. The relationship between the complement activity of the patients with coronary heart disease and the carrying of MICA*008 gene was studied. Methods: 1. first, the human red blood cells were replaced by the sheep red blood cells, and the Rabbit anti human erythrocyte antibody replaced the Rabbit anti sheep erythrocyte antibody (hemolysin) to carry out the hemolysis test. It was proved that the different red cell concentration and plasma concentration were able to release the hemoglobin after the red blood cells were destroyed. The most suitable concentration of various reagents in hemolysis was determined by the CH50 multi tube hemolysis test, and a single tube hemolysis colorimetric method was created. Meanwhile, the accuracy and stability of the method was verified by linear and repeated tests..2. was used to detect the complement activity in plasma and serum by CH50 standard method, and the single tube hemolysis colorimetric method was used. The method was used to detect the complement activity in the same plasma with different human red cells as a hemolytic indicator system. The feasibility of single tube hemolysis colorimetric method was further verified by.3. and single tube hemolysis colorimetric assay to detect the complement activity of the same plasma by CH50 method and single tube hemolysis colorimetry. The correlation.4. of two methods was tested for the detection of ICU patients by single tube hemolysis test. The total complement activity of the patients with heart disease and health checkup proves that the method has certain clinical value. At the same time, PCR technique is used to detect the MICA*008 gene in patients with coronary heart disease and healthy people, and the relationship between the complement activity and the MICA*008 gene of the patients with coronary heart disease is analyzed. 1. the results of the results are the use of human red blood cells and Rabbit anti human erythrocyte antibodies. The results showed that the effect of different concentrations of red blood cells on the degree of hemolysis was lower than that of the plasma at different concentrations. At the same time, the effect of the different concentration of red blood cells on the efficiency of hemoglobin release after the broken red cells was 20%, so the effect of the different concentration of red blood cells on the release of hemoglobin was the lowest. The content of each component in the ninth tube (20% plasma concentration) of the 20% red cell suspension was used as a reference for the proportion of the components in the single tube hemolysis colorimetric method. The complement activity of the plasma was detected by the single tube hemolysis colorimetry, and the complement activity of the same plasma was measured for 5 days. The results showed that the plasma was supplemental at different concentrations. The total body activity was linearly distributed and the variation coefficient of the complement activity value was CV=11.3% for 5 days. It showed that the single tube hemolysis colorimetric test had certain accuracy and stability..2. was used to detect the complement activity in plasma and serum by CH50 standard method. The complement activity in plasma and serum was the same, so plasma could be used in plasma. The complement activity was measured and the complement detection technique was optimized. At the same time, the complement activity in the same plasma was detected by different human red cells as a hemolytic indicator system. The complement activity in the bleeding pulp was approximately the same (P0.05), indicating that different erythrocytes would not affect the complement content in the plasma and the single tube hemolysis. The method is feasible..3. the hemolysis o value measured in this method is linearly related to the value of the complement activity measured by the CH50 hemolysis method. It shows that the single tube hemolysis colorimetric method has a significant correlation with the CH50 method. The single tube hemolysis colorimetric method can replace the CH50 hemolysis method to detect the complement activity of the total complement active.4.ICU patients and the coronary heart disease patients. The single tube hemolysis colorimetric method has a certain clinical value below the complement activity of the health examiners. The MICA*008 gene distribution in the patients with coronary heart disease and the healthy persons is roughly the same, and the complement level of the patients with coronary heart disease is not significantly associated with the MICA*008. Conclusion: 1. single tube hemolysis has good accuracy for the detection of complement activity. Sex, stability and feasibility, and have a significant correlation with the CH50 method, this method further optimizes the CH50 hemolysis test, which has certain clinical application value of.2.ICU patients and coronary heart disease patients with lower complement activity, indicating the immune function disorder of the body of ICU patients, the body of the patients with coronary heart disease may be in the stage of.3. coronary heart disease in the stage of inflammation. There was no significant correlation between complement level and MICA*008 gene.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R541.4;R446.1
本文編號(hào)：2156294
[Abstract]:Objective: on the basis of CH50 hemolysis, we use human erythrocytes to replace sheep red blood cells and convert multi tube method into single tube method to create a new method to detect the total complement activity of plasma complement, which is simple and stable and reliable. At the same time, this method is used to detect the total complement activity of ICU patients, coronary heart disease patients and health examiners. The relationship between the complement activity of the patients with coronary heart disease and the carrying of MICA*008 gene was studied. Methods: 1. first, the human red blood cells were replaced by the sheep red blood cells, and the Rabbit anti human erythrocyte antibody replaced the Rabbit anti sheep erythrocyte antibody (hemolysin) to carry out the hemolysis test. It was proved that the different red cell concentration and plasma concentration were able to release the hemoglobin after the red blood cells were destroyed. The most suitable concentration of various reagents in hemolysis was determined by the CH50 multi tube hemolysis test, and a single tube hemolysis colorimetric method was created. Meanwhile, the accuracy and stability of the method was verified by linear and repeated tests..2. was used to detect the complement activity in plasma and serum by CH50 standard method, and the single tube hemolysis colorimetric method was used. The method was used to detect the complement activity in the same plasma with different human red cells as a hemolytic indicator system. The feasibility of single tube hemolysis colorimetric method was further verified by.3. and single tube hemolysis colorimetric assay to detect the complement activity of the same plasma by CH50 method and single tube hemolysis colorimetry. The correlation.4. of two methods was tested for the detection of ICU patients by single tube hemolysis test. The total complement activity of the patients with heart disease and health checkup proves that the method has certain clinical value. At the same time, PCR technique is used to detect the MICA*008 gene in patients with coronary heart disease and healthy people, and the relationship between the complement activity and the MICA*008 gene of the patients with coronary heart disease is analyzed. 1. the results of the results are the use of human red blood cells and Rabbit anti human erythrocyte antibodies. The results showed that the effect of different concentrations of red blood cells on the degree of hemolysis was lower than that of the plasma at different concentrations. At the same time, the effect of the different concentration of red blood cells on the efficiency of hemoglobin release after the broken red cells was 20%, so the effect of the different concentration of red blood cells on the release of hemoglobin was the lowest. The content of each component in the ninth tube (20% plasma concentration) of the 20% red cell suspension was used as a reference for the proportion of the components in the single tube hemolysis colorimetric method. The complement activity of the plasma was detected by the single tube hemolysis colorimetry, and the complement activity of the same plasma was measured for 5 days. The results showed that the plasma was supplemental at different concentrations. The total body activity was linearly distributed and the variation coefficient of the complement activity value was CV=11.3% for 5 days. It showed that the single tube hemolysis colorimetric test had certain accuracy and stability..2. was used to detect the complement activity in plasma and serum by CH50 standard method. The complement activity in plasma and serum was the same, so plasma could be used in plasma. The complement activity was measured and the complement detection technique was optimized. At the same time, the complement activity in the same plasma was detected by different human red cells as a hemolytic indicator system. The complement activity in the bleeding pulp was approximately the same (P0.05), indicating that different erythrocytes would not affect the complement content in the plasma and the single tube hemolysis. The method is feasible..3. the hemolysis o value measured in this method is linearly related to the value of the complement activity measured by the CH50 hemolysis method. It shows that the single tube hemolysis colorimetric method has a significant correlation with the CH50 method. The single tube hemolysis colorimetric method can replace the CH50 hemolysis method to detect the complement activity of the total complement active.4.ICU patients and the coronary heart disease patients. The single tube hemolysis colorimetric method has a certain clinical value below the complement activity of the health examiners. The MICA*008 gene distribution in the patients with coronary heart disease and the healthy persons is roughly the same, and the complement level of the patients with coronary heart disease is not significantly associated with the MICA*008. Conclusion: 1. single tube hemolysis has good accuracy for the detection of complement activity. Sex, stability and feasibility, and have a significant correlation with the CH50 method, this method further optimizes the CH50 hemolysis test, which has certain clinical application value of.2.ICU patients and coronary heart disease patients with lower complement activity, indicating the immune function disorder of the body of ICU patients, the body of the patients with coronary heart disease may be in the stage of.3. coronary heart disease in the stage of inflammation. There was no significant correlation between complement level and MICA*008 gene.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R541.4;R446.1
【相似文獻(xiàn)】
相關(guān)期刊論文 前10條
1 歐陽(yáng)錄明;總補(bǔ)體活性檢測(cè)實(shí)驗(yàn)技術(shù)改革[J];郴州醫(yī)專學(xué)報(bào);2000年02期
2 謝秩勛,張萍,李素華,朱國(guó)標(biāo);高溫環(huán)境對(duì)補(bǔ)體活性的影響[J];西南國(guó)防醫(yī)藥;1994年02期
3 謝軍,周莉,謝冰,朱國(guó)標(biāo),謝秩勛;軍用固體運(yùn)動(dòng)飲料對(duì)機(jī)體補(bǔ)體活性影響的實(shí)驗(yàn)研究[J];西南國(guó)防醫(yī)藥;2003年01期
4 謝fs子;;血清總補(bǔ)體活性測(cè)定(摘要)[J];醫(yī)衛(wèi)通訊;1980年02期
5 衣承東;;大手術(shù)后和ARDS病人的補(bǔ)體活性及肺泡毛細(xì)血管通透性增加[J];國(guó)外醫(yī)學(xué).創(chuàng)傷與外科基本問(wèn)題分冊(cè);1988年04期
6 陳晏珍;;處于成人呼吸窘迫綜合征危險(xiǎn)時(shí)的補(bǔ)體活性[J];國(guó)外醫(yī)學(xué).呼吸系統(tǒng)分冊(cè);1985年04期
7 方蕾;檢測(cè)補(bǔ)體活性的方法[J];國(guó)外醫(yī)學(xué)(免疫學(xué)分冊(cè));1994年01期
8 張進(jìn)友;王秀文;;瓊脂糖凝膠內(nèi)溶血法定量檢測(cè)人血清補(bǔ)體活性的研究[J];首都醫(yī)學(xué)院學(xué)報(bào);1993年02期
9 馮偉華,何新全;總補(bǔ)體活性（CH50）的簡(jiǎn)易測(cè)定法——公式計(jì)算法[J];華西醫(yī)學(xué);1996年02期
10 仝連信,姜玉祥,梁曙光;脂質(zhì)體免疫檢測(cè)法測(cè)定血清總補(bǔ)體活性臨床價(jià)值的探討[J];牡丹江醫(yī)學(xué)院學(xué)報(bào);2000年01期
相關(guān)碩士學(xué)位論文 前1條
1 董蕊銳;基于自身紅細(xì)胞構(gòu)建的溶血系統(tǒng)檢測(cè)總補(bǔ)體活性及其與MICA基因多態(tài)性的關(guān)系[D];大連醫(yī)科大學(xué);2016年
，本文編號(hào)：2156294
本文鏈接：http://sikaile.net/kejilunwen/jiyingongcheng/2156294.html
最近更新
教材專著
