本文選題：熒光微球 + 血液篩查��； 參考：《西北大學(xué)》2005年碩士論文

【摘要】：因輸血導(dǎo)致感染乙肝、丙肝、艾滋、梅毒等疾病的現(xiàn)象在當(dāng)今社會已經(jīng)越來越普遍,引起國家和社會的高度重視。目前最主要的任務(wù)就是嚴(yán)格控制對外來血源的篩查,而傳統(tǒng)的ELISA方法檢測血液傳播疾病的方法仍在不同方面存在局限性,如檢測通量低、需時長、靈敏度較低等。本研究采用流式熒光微球技術(shù),應(yīng)用ELISA雙抗體夾心法和間接法原理以及生物素-親和素系統(tǒng),目的在于建立一種高通量、快速、靈敏檢測HBV、HCV等病毒或病菌血清標(biāo)志物的方法。 具體研究工作可分為三部分: 1.用Hb-PcAb和羊抗人IgG優(yōu)化抗體生物素化條件。 2.用Hb和人IgG建立抗原(抗體)與熒光微球的偶聯(lián)條件。 3.以聚苯乙烯羧基熒光微球?yàn)楣滔噍d體,將HBsAg-McAb-S6和HCV Core Ag與不同編號的聚苯乙烯羧基熒光微球偶聯(lián);應(yīng)用ELISA雙抗體夾心法和間接法原理在Luminex-100~(TM)流式熒光微球檢測系統(tǒng)下檢測人血清中的HBsAg和HCV-Ab。 結(jié)果表明抗體在以PBS作為第一次透析液、DMF作為BNHS溶劑的條件下生物素化效果較好;抗體生物素化后活性保持較好,生物素化效率較高�？乖�/抗體分別在100mM MES(PH6.0)和50mM MES(PH5.0)中,濃度為100μg/ml條件下與熒光微球的偶聯(lián)效率最高。熒光微球檢測128例HBV患者血清中HBsAg的特異性和靈敏度均為100%,大大高于ELISA檢測的特異性(71.43%)和靈敏度(66.67%),顯示了熒光微球技術(shù)檢測的優(yōu)越性;熒光微球檢測74例HCV患者血清中HCV-Ab的靈敏度為89.19%,檢測方法還有待于提高。
[Abstract]:Infection with hepatitis B, hepatitis C, AIDS, syphilis and other diseases caused by blood transfusion has become more and more common in today's society. At present, the most important task is to strictly control the screening of foreign blood sources. However, the traditional ELISA method for detecting blood-borne diseases still has some limitations in different aspects, such as low detection flux, long detection time, low sensitivity and so on. In this study, ELISA double antibody sandwich method and indirect method and biotin avidin system were used to establish a high throughput, rapid and sensitive method for the detection of HBV virus or virus markers. The specific research can be divided into three parts: 1. Hb-PcAb and sheep anti-human IgG were used to optimize the biotinylation conditions. 2. The coupling conditions between antigens (antibodies) and fluorescent microspheres were established by HB and human IgG. 3. Using polystyrene carboxyl fluorescent microspheres as solid phase carrier, HBsAg-McAb-S6 and HCV Core Ag were coupled with polystyrene carboxyl fluorescent microspheres with different numbers. ELISA double antibody sandwich method and indirect method were used to detect HBsAg and HCV-Abin in human serum under the Luminex-100T TM flow fluorescence microsphere detection system. The results showed that the biotinylation effect of antibody was better when PBS was used as the first dialysate and the biotinylation efficiency was higher. When the concentration of antigen / antibody was 100 渭 g/ml, the coupling efficiency with fluorescent microspheres was the highest. The specificity and sensitivity of fluorescent microspheres in the detection of HBsAg in serum of 128 patients with HBV were 100, which was much higher than that of ELISA (71.43) and the sensitivity was 66.67%, which showed the superiority of fluorescent microspheres in the detection of HBsAg. The sensitivity of fluorescent microspheres in the detection of HCV-Ab in 74 patients with HCV was 89.19, and the detection method needed to be improved.
【學(xué)位授予單位】：西北大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號】：R373

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 趙化陽;陳立超;李偉;白玉;尹良宏;盧勝明;王小珂;;流式熒光微球技術(shù)在實(shí)驗(yàn)動物質(zhì)量控制中的應(yīng)用[J];實(shí)驗(yàn)動物科學(xué);2012年01期

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本文編號：1931989