本文選題：納米金棒　切入點(diǎn)：血吸蟲病　出處：《大理大學(xué)》2017年碩士論文

【摘要】：目的納米技術(shù)目前已成為多種學(xué)科研究的熱點(diǎn)問題和前沿方向,納米金作為一種性能優(yōu)良的納米材料在疾病的早期診斷、檢測以及治療方面得到廣泛研究。本研究對納米金棒的制備條件進(jìn)行了優(yōu)化,選擇最具有所需光學(xué)性質(zhì)的納米金棒,并成功將其功能化,制備了實(shí)驗(yàn)室條件下可針對日本血吸蟲病檢測的高特異性、敏感性的納米金生物探針。方法通過種子介導(dǎo)的生長法聚合納米金棒,通過對增長媒質(zhì)中不同成分的比例、反應(yīng)時(shí)間、溫度等控制,系統(tǒng)的研究納米金棒最佳的聚合條件,最大程度的提高單個(gè)納米顆粒的生物功能,聚合具有所需的形貌和波長可控的納米金棒。制備日本血吸蟲尾蚴和成蟲可溶性抗原,并將抗原與納米金棒結(jié)合,構(gòu)建NanoSPR生物探針,將NanoSPR生物探針功能化,通過表面等離子共振峰的LSPR峰變化來探索抗原包被的最佳濃度。將功能化的NanoSPR生物探針檢測不同濃度梯度的感染日本血吸蟲陽性血清抗體,并與ELISA檢測結(jié)果做對照分析研究,探索日本血吸蟲NanoSPR生物芯片的高特異性。將功能化的NanoSPR生物探針檢測感染日本血吸蟲前1-8周的兔血清抗體,并與ELISA檢測結(jié)果做對照分析研究,探索日本血吸蟲NanoSPR生物芯片的高敏感性。結(jié)果通過對GNRs在制備過程中還原劑、活性劑、溫度等條件的優(yōu)化,系統(tǒng)的研究了GNRs的最佳實(shí)驗(yàn)制備條件。在增長媒質(zhì)中CTAB的最佳濃度為0.095mol/L,AA的最佳濃度為0.640m mol/L,AgNO3的最佳濃度為72μmol/L,種子的最佳生長溫度為28°C,種子溶液在增長媒質(zhì)中生長12h基本趨于穩(wěn)定,各項(xiàng)指標(biāo)基本不再發(fā)生變化。在成功制備NanoSPR生物探針后,同過對NanoSPR生物探針不同的實(shí)驗(yàn)分析得出:包被濃度為20μg/m L時(shí)包被前后LSPR峰出現(xiàn)了最大程度的紅移達(dá)到18nm且峰形正常,確定為最佳包被濃度;對比包被兩種不同抗原的GNRs的檢測結(jié)果,其中NanoSPR-SCA生物探針相對于NanoSPR-AWA的識(shí)別日本血吸蟲感染早期的血清具有一定的優(yōu)勢;相比于ELISA檢測結(jié)果的對比發(fā)現(xiàn),通過納米金標(biāo)記對抗原抗體反應(yīng)產(chǎn)生的放大作用,使NanoSPR-AWA生物探針對血清中血吸蟲抗體的檢測更具有靈敏性,最低檢測限可達(dá)到稀釋倍數(shù)的80倍。相比于傳統(tǒng)的ELISA檢測法,NanoSPR生物探針針對日本血吸蟲感染早期的血清體現(xiàn)出了更強(qiáng)的識(shí)別能力。結(jié)論利用種子生長法通過探索最佳CTAB、AgNO3、AA等試劑濃度和影響金棒制備因素的實(shí)驗(yàn)條件,制備了長徑比較高、等離子體共振峰位置和強(qiáng)度可有效調(diào)控的納米金棒。并對基于納米金標(biāo)記的免疫傳感器NanoSPR生物探針進(jìn)行了實(shí)驗(yàn)室階段的基礎(chǔ)研究,證實(shí)基于納米金標(biāo)記的傳感器是一種可以針對日本血吸蟲病具有更高特異性,更靈敏的檢測方法和工具,并為豐富日本血吸蟲病的檢測方法提供了技術(shù)支持的新思路。
[Abstract]:Objective Nano-technology has become a hot topic and a leading direction in many disciplines. As an excellent nano-material, nano-gold can be used in the early diagnosis of disease. In this study, the preparation conditions of gold nanorods were optimized, and the nanocrystalline gold rods with the most needed optical properties were selected, and the nanocrystalline gold rods were successfully functionalized. A highly specific and sensitive nanoscale gold bioprobe for detection of schistosomiasis japonicum was prepared under laboratory conditions. Methods the nanocrystalline gold rods were polymerized by seeded growth method and by the proportion of different components in the growth medium. Under the control of reaction time, temperature and so on, the optimum polymerization conditions of nanocrystalline gold rods were studied systematically, and the biological function of single nanoparticles was maximized. The soluble antigens of cercariae and adults of Schistosoma japonicum were prepared by polymerizing the gold nanorods with the desired morphology and wavelength. The NanoSPR biological probes were constructed by combining the antigens with gold nanorods, and the NanoSPR bioprobes were functionalized. The best concentration of antigen coating was explored by the change of LSPR peak of surface plasmon resonance peak. The positive serum antibody of different concentration gradient infected Schistosoma japonicum was detected by functional NanoSPR biological probe and compared with the result of ELISA detection. To explore the high specificity of NanoSPR biochip of Schistosoma japonicum, the functional NanoSPR bioprobe was used to detect the serum antibodies of rabbits before 1 to 8 weeks of infection with Schistosoma japonicum, and the results were compared with the results of ELISA. To explore the high sensitivity of NanoSPR biochip of Schistosoma japonicum. Results by optimizing the conditions of reducing agent, active agent, temperature and so on during the preparation of GNRs, The optimum preparation conditions of GNRs were systematically studied. The optimum concentration of CTAB was 0.095 mol / L, the optimum concentration of CTAB was 0.640 mmol / L, the optimum concentration of Agno _ 3 was 72 渭 mol / L, the optimum growth temperature of seed was 28 擄C, and the seed solution was stable in growth medium for 12 h. After the successful preparation of NanoSPR biological probe, different experimental analysis on NanoSPR bioprobe showed that the maximum redshift of the LSPR peak before and after the coating was 20 渭 g / mL, and the peak shape was normal, and the maximum red shift of the LSPR peak was observed when the coating concentration was 20 渭 g / mL. Compared with the results of GNRs coated with two different antigens, NanoSPR-SCA biological probe has some advantages over NanoSPR-AWA in identifying the early serum of Schistosoma japonicum infection, and compared with the results of ELISA detection. Through the amplification of the antigen-antibody reaction by nano-gold labeling, the NanoSPR-AWA biological probe is more sensitive to the detection of schistosomiasis antibody in serum. The minimum detection limit can reach 80 times of dilution multiple. Compared with the traditional ELISA method, the detection of ELISA biological probe for early infection of Schistosoma japonicum showed a stronger ability to recognize the serum. Conclusion the seed growth method can be used to explore the method of detection of Schistosoma japonicum in the early stage of infection. The optimum concentration of CTABX Agno _ 3H _ 3AA and the experimental conditions affecting the preparation of gold rods were studied. Nanocrystalline gold rods with high length and high plasma resonance peak position and intensity were prepared. The basic research of NanoSPR biosensor based on nanoscale gold labeling was carried out in laboratory. It is proved that the sensor based on gold nanoparticles is a more specific and sensitive method and tool for detection of schistosomiasis japonicum, and provides a new idea for enriching the detection methods of schistosomiasis japonicum.
【學(xué)位授予單位】：大理大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R532.21

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