Jiang Lixiang, Luo Jing, Dong Wenjie, Wang Chengmin, Jin Wen, Xia Yuetong, Wang Haijing, Ding Hua, Jiang Long, He Hongxuan
National Research Center for Wildlife Born Diseases, Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China.
National Research Center for Wildlife Born Diseases, Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
J Virol Methods. 2015 Jul;219:38-45. doi: 10.1016/j.jviromet.2015.03.013. Epub 2015 Mar 26.
H5N1 avian influenza has caused serious economic losses as well as posed significant threats to public health, agriculture and wildlife. It is important to develop a rapid, sensitive and specific detection platform suitable for disease surveillance and control. In this study, a highly sensitive, specific and rapid biosensor based on polydiacetylene was developed for detecting H5 influenza virus. The polydiacetylene based biosensor was produced from an optimized ratio of 10,12-pentacosadiynoic acid and 1,2-dimyristoyl-sn-glycero-3-phosphocholine, with the anti-H5 influenza antibody embedded onto the vesicle surface. The optimized polydiacetylene vesicle could detect H5 influenza virus sensitively with a detection limit of 0.53 copies/μL, showing a dramatic blue-to-red color change that can be observed directly by the naked eye and recorded by a UV-vis spectrometer. The sensitivity, specificity and accuracy of the biosensor were also evaluated. The sensor could specifically differentiate H5 influenza virus from H3 influenza virus, Newcastle disease virus and porcine reproductive and respiratory syndrome virus. Detection using tracheal swabs was in accord with virus isolation results, and comparable to the RT-PCR method. These results offer the possibility and potential of simple polydiacetylene based bio-analytical method for influenza surveillance.
H5N1禽流感已造成严重经济损失，并对公众健康、农业和野生动物构成重大威胁。开发一个适用于疾病监测和控制的快速、灵敏且特异的检测平台非常重要。在本研究中，开发了一种基于聚二乙炔的高灵敏、特异且快速的生物传感器用于检测H5流感病毒。该基于聚二乙炔的生物传感器由10,12-二十五碳二炔酸和1,2-二肉豆蔻酰-sn-甘油-3-磷酸胆碱的优化比例制备而成，抗H5流感抗体包埋在囊泡表面。优化后的聚二乙炔囊泡能够灵敏地检测H5流感病毒，检测限为0.53拷贝/微升，呈现出肉眼可直接观察到的显著的蓝到红颜色变化，并可用紫外可见光谱仪记录。还评估了该生物传感器的灵敏度、特异性和准确性。该传感器能够特异性地区分H5流感病毒与H3流感病毒、新城疫病毒和猪繁殖与呼吸综合征病毒。使用气管拭子进行检测与病毒分离结果一致，且与RT-PCR方法相当。这些结果为基于聚二乙炔的简单生物分析方法用于流感监测提供了可能性和潜力。
