【作者】 杨会娜；

【导师】 蔡家利；

【作者基本信息】 重庆理工大学 ， 微生物与生化药学， 2011， 硕士

【摘要】 禽流感(avian influenza)是由A型流感病毒引起的一种禽类传染性疾病。近年来多项研究已表明,这类病毒可感染人类引起发病,称为禽流感病毒感染。禽流感给人类带来了巨大的经济损失和健康威胁。据中国动物卫生与流行病学中心国际兽医事物综合分析室公布的数据,2010年全球通报发生的诸多重大动物疫情中,禽流感疫情仍排在第一位,这就说明当前禽流感的疫情仍然严重,仍需要投入大量的人力物力,抑制或缓解这一疫情。传统的禽流感检测方法,如血凝和血凝抑制,对设备的要求不高,经济成本低,但对操作者的熟练程度要求很高,尤其对试验结果的分析和判定,不似其他诸如酶标仪等的判断结果具有直观性和机械性,而核酸检测技术中所用到的仪器,虽精确但大都价格昂贵,不适宜在基层做大量推广。液体电谐振传感仪在诊断禽流感上的应用研究,将传统的血凝和血凝抑检测方法之间作对比,旨在建立一种新的禽流感检测技术和开发新的检测仪器,达到以低廉的成本可以便捷的检测禽流感的目的,能够应用于检验检疫部门、防疫部门和医院。本课题研究的液体电谐振传感仪检测禽流感技术是属于生物传感仪技术的一种,其原理是利用抗原与抗体反应的特异性和和石英谐振换能器对表面质量负载变化的高度敏感性相结合,通过测定发生生化反应的液体电谐振频率的变化,来表征试验结果的一种新型压电免疫传感器的检测方法。将液体电谐振传感仪应用于禽流感的检测具有检测速度快、经济、灵敏性高等优点。并且,检测中用到的反应芯片按照规定的程序清洗后,可重复使用,适合大规模测试。一旦疫情发生,可做到有效地检测。本课题首先采用鸡胚培养法将试验中要大量用到的AIV标毒进行增殖,得到约70ml的AIV病毒尿囊液,再采用醛化红细胞吸附法对病毒进行纯化,得到两管病毒,1号管约6ml,2号管约5ml,同时紫外分光光度法计算得知1号管的浓度为18.31mg/ml,2号浓度为15.35mg/ml。将成熟的血凝试验和血凝抑制试验与液体电谐振试验作对比,当待测病毒稀释度在其效价范围内时,并且其谐振频率最小值小于300000Hz,液体电谐振传感仪测得的平均频移值与其稀释度的自然对数呈线性关系,且线性关系系数R2值接近1,判为血凝阳性。当待测病毒的稀释度低于效价时,其谐振频率大于300000 Hz,同时平均频移值与其稀释度的自然对数线性关系有较大的偏差,则此时的待测液的效价判为血凝阴性。更多还原

【Abstract】 Avian flu (avian influenza) is caused by A-type influenza A virus avian infectious diseases. In recent years, several studies have shown that this virus can infect humans cause disease, called avian influenza. Avian flu to humans has brought huge economic losses and health threats. According to the China Animal Health and Epidemiology Center, Room comprehensive analysis of international veterinary matters data published in 2010, many of the world informed of major animal epidemic occurred, the bird flu in the first row, which indicates that the current outbreak of avian flu remains a serious still requires a lot of manpower and resources, inhibit or alleviate the disease.The traditional avian flu detection methods, such as hemagglutination and hemagglutination inhibition, less demanding on the equipment, the economy, low cost, but the proficiency of the operator demanding, especially for the analysis and determine the results, unlike other countries such as microplate reader judge the outcome of such a visual and mechanical detection of the nucleic acid used in the instrument, although accurate but most expensive, is not appropriate to do a lot of promotion at the grassroots level.Liquor Electrical Resonance Sensor in the diagnosis of avian influenza on the instrument the application of research to the traditional method of hemagglutination and hemagglutination inhibition test production compared to the establishment of a new bird flu detection techniques and the development of new detection equipment, to achieve a low The costs can be convenient for the purpose of detection of avian influenza, can be used in the inspection and quarantine departments, epidemic prevention departments and hospitals. Liquor Electrical Resonance Sensor with the detection of avian flu rapid test, economic, and sensitivity advantages. Moreover, detection of the reaction used in accordance with the procedure chips after washing, reusable, suitable for large-scale testing. Once the outbreak can be effectively detected.The research of liquid electric resonance sensor technology is avian influenza detected by a biological sensor technology, a kind of instrument, the principle is the use of antigen specificity and antibody response resonant quartz transducer and the surface quality of the load of the height variation sensitivity of the combination of biochemical reactions occur by measuring the resonant frequency changes liquids, to characterize the results of a novel piezoelectric immunosensor detection methods.First, the subject of embryo culture method used to test a large number of standard drugs used for the proliferation of AIV, AIV virus by about 70ml of allantoic fluid, and then red blood cells using formaldehyde adsorption of the virus was purified by two viruses, tube 1 about 6ml, tube 2 about 5ml, calculated by UV spectrophotometry at the same time that the concentration of tube 1 is 18.31mg/ml, tube 2 concentration 15.35mg/ml.Will mature hemagglutination and hemagglutination inhibition test and compare the liquid electric resonance experiment, when tested in the virus titer dilution range, and its resonant frequency is less than the minimum 300000Hz, liquids Measuring resonant sensor The mean frequency shift value and its natural logarithm of dilution is linear, and linear correlation coefficient R2 value close to 1, ruled hemagglutination positive. When the analyte is lower than the dilution of the virus titer, its resonant frequency is greater than 300000 Hz, while the average frequency shift value and its dilution of the linear relationship between the natural logarithm of a large deviation, the fluid under test at this time hemagglutination titer ruled negative.更多还原