H5NI型禽流感病毒纳米量子点快速检测方法的建立

Studies on quantum Dots for Quick Detection of H5N1 subtype Avian Influenza Virus

本文介绍了一种用CdSe量子点标记技术用于H5NI型禽流感病毒检测的方法。采用CdSe量子点标记鸡抗AIVIgG,并对标记的抗体量子点复合产物进行了纯化。H5N1型禽流感病毒单克隆抗体作为包被抗体,CdSe量子点标记的鸡抗AIVIgG作为检测抗体,建立了纳米量子点应用于禽流感病毒快速检测的方法。此方法的最佳检测组织为气管和肺,整个检测过程只需3小时。实验结果表明,对已知的阳性样品其敏感性比血凝试验要高出4倍以上,与新城疫病毒、鸭瘟病毒和鸭肝炎病毒等无交叉反应。对65份病料进行检测,结果有5份阳性病例,60份阴性病例,与鸡胚分离法作为比对,结果符合率为98.46%,说明此方法具有较好的特异性和灵敏度,该方法操作简单、检测快速快,在进出口检疫方面有良好的应用前景。

A test method using quantum dots conjugation was developed for the detection of H5N1 subtype avian influenza viros（AIV）.CdSe quantum dots（QDs） was used to label chicken anti-AIV IgG, and the production of the CdSe quantum Dots labeled chicken anti-AIV IgG was purified. The purified conjugate of chicken anti-AIV IgG labeled with CdSe quantum dots used as second antibody, and anti-H5N1 subtype AIV mAhs was coated on the ELISA plate. Trachea and lung were the most suitable tissues for the diagnosis of AIV for the quantum dots test. The entie test takes only 3 hours. The sensitivity of this method used to detect AIV positive sample is 4 times higher than that by hemagglutination test. No cross reaction was observed with Newcastle disease viros（NDV）,duck plague viros（DPV） and duck hepatitis（DVH）.65 clinical samples were detected by this method, and 5 samples were positive, and 60 samples were negative. Compared with virus isolation, this diagnostic accordance rate is 98.46 %. The QDs test method established in this study had been demonstrated to he a specific, sensitive and rapid test for the diagnosis of AIV, and it offers a promising possibility to he used in entry-exit inspection and quarantine.