本文選題：萊克多巴胺 + 免疫層析�。� 參考：《暨南大學》2012年碩士論文

【摘要】：目的：依據(jù)免疫競爭層析方法的原理，建立萊克多巴胺（RAC）熒光微球免疫層析快速檢測方法。研制萊克多巴胺熒光微球免疫層析試紙條，用于豬尿中萊克多巴胺殘留的檢測。 方法：采用飽和硫酸銨沉淀法純化抗RAC單抗小鼠腹水，并用BCA試劑盒測定其抗體蛋白濃度。以EDC作為偶聯(lián)劑，將本實驗室制備的抗RAC單抗偶聯(lián)到商用聚苯乙烯熒光微球表面，合成出熒光微球抗體復(fù)合物。將標記好的復(fù)合物噴涂于結(jié)合墊上；采用活性酯法合成RAC-BSA檢測抗原并將其包被在NC膜表面作為檢測線(T線)；羊抗鼠IgG包被在硝酸纖維素（NC）膜表面作為質(zhì)控線(C線)；篩選并優(yōu)化復(fù)溶液及樣品墊預(yù)處理液中的各項成分及濃度。將結(jié)合墊、吸水紙和處理過的樣品墊最后組裝成試紙條。試紙條檢測過程中RAC-BSA與待測樣品中RAC競爭結(jié)合有限的熒光微球標記RAC單抗，如果樣品中的RAC含量多則其結(jié)合熒光微球標記的抗體多，則結(jié)合到T線的熒光微球抗體復(fù)合物越少，于450nm激發(fā)光下觀察，則T線的熒光強度越低，因此可根據(jù)熒光強度的強弱檢測判斷樣品中RAC的多寡。 結(jié)果：合成出檢測抗原RAC-BSA。合成得到了熒光微球抗體復(fù)合物，該復(fù)合物與檢測抗原有很好的結(jié)合，且與BSA不存在非特異性結(jié)合。建立了RAC熒光快速檢測方法，制備出了RAC熒光微球免疫層析試紙條，該試紙條檢測豬尿中RAC的殘留，，靈敏度最低值為2ngmL~(-1)，與克倫特羅（CL），沙丁胺醇（SAL）等類似物無交叉反應(yīng)。 結(jié)論：本研究建立的萊克多巴胺熒光微球免疫層析快速檢測技術(shù)操作便捷，穩(wěn)定可靠，靈敏度高，可作為豬尿中萊克多巴胺殘留現(xiàn)場檢測和監(jiān)控的有效手段。
[Abstract]:Aim: to establish a rapid immunochromatographic method for detection of ractopamine rac fluorescent microspheres according to the principle of immunocompetitive chromatography. The ractopamine fluorescent microspheres immunochromatographic strip was developed for the detection of ractopamine residues in pig urine. Methods: the mouse ascites of anti RAC monoclonal antibody were purified by saturated ammonium sulfate precipitation method and the concentration of antibody protein was determined by BCA kit. Using EDC as coupling agent, the anti RAC monoclonal antibody prepared in our laboratory was coupled to the surface of commercial polystyrene fluorescent microspheres to synthesize fluorescent microspheres antibody complexes. Spraying the labeled composite onto the binding pad; RAC-BSA detection antigen was synthesized by active ester method and coated on the surface of NC membrane as T line; goat anti rat IgG coating on nitrocellulose membrane surface was used as quality control line C line; complex solution and sample pad pretreatment were selected and optimized. The composition and concentration of the liquid. Combine mats, absorbent paper and treated mats to form test strips. In the process of test strip detection, RAC-BSA competes with RAC in the test sample to bind to the limited fluorescent microsphere labeled RAC McAb. If the sample contains more RAC, it binds to the fluorescent microsphere antibody more, then the less fluorescent microsphere antibody complex binds to the T line. Observed under 450nm excitation light, the fluorescence intensity of T line is lower. Therefore, the amount of RAC in the sample can be judged by the detection of fluorescence intensity. Results: the antigen RAC-BSA was synthesized. The fluorescent microsphere antibody complex was synthesized, which binds well to the detection antigen and has no nonspecific binding to BSA. A rapid RAC fluorescence detection method was established, and a RAC fluorescent microsphere immunochromatographic strip was prepared. The test strip was used to detect RAC residues in porcine urine. The lowest sensitivity was 2ngmLnLX, and there was no cross reaction with clenbuterol and clenbuterol (clenbuterol) and the other analogs, such as clenbuterol or clenbuterol. Conclusion: the rapid detection technique of ractopamine fluorescence microsphere immunochromatography is easy to operate, stable, reliable and sensitive. It can be used as an effective method for detection and monitoring of ractopamine residues in pig urine.
【學位授予單位】：暨南大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R392.1

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