摘要
目的:建立高灵敏检测蓝舌病毒VP7蛋白的生物条形码检测方法。方法:制备VP7蛋白的多抗及特异DNA链标记的金纳米颗粒探针(NP)和VP7蛋白单抗标记的磁性微球探针(MMP),形成MMP-VP7蛋白-NP三明治复合物后,再利用去杂交将NP探针上标记的DNA链释放出来,通过PCR或芯片检测方法鉴定释放的DNA链,确定VP7蛋白的存在。结果:建立了蓝舌病毒VP7蛋白的生物条形码检测体系,检测灵敏度可达10fg/mL,为常规ELISA检测的106倍。结论:为发展高灵敏度的蓝舌病毒生物条形码检测试剂盒鉴定了基础。
Objective： To establish a high sensitive bio-bar codes assay method for detecting bluetongue virus（BTV） VP7 protein. Methods： Nanoparticle（NP） probes encoded with DNA that wass unique to VP7 protein, polyclonal antibodies agaist VP7 protein and magnetic microparticle （MMP） probes with monoclonal antibodies that bind VP7 protein specifically were prepared. After forming MMP-VP7-NP sandwich complex, dehybridization of the oligonucleotides on the NP surface allows the determination of the presence of VP7 protein by identifying the oligonucleotide sequence released from the NP through PCR or chip-based detection. Results： The bio-bar codes assay system for detecting VP7 protein was established, and the sensitivity limit was about 10 fg/mL, comparable clinically accepted conventional ELISA assays for detecting the same targe, six orders of magnitude less sensitive than what was observed with this method. Conclusion： This study lays a foundation for establishing high sensitive BTV bio-bar codes assay kit.
出处
《生物技术通讯》
CAS
2008年第5期697-700,共4页
Letters in Biotechnology
基金
国家高技术研究发展计划(2006AA10446)
