| 利用生物条形码技术对蓝舌病毒VP7蛋白进行微量检测 |
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| 引用本文： | 张立营,张红,杨姝,吕茂民,尹惠琼,张改平,李德雪,章金刚. 利用生物条形码技术对蓝舌病毒VP7蛋白进行微量检测[J]. 生物技术通讯, 2008, 19(5): 697-700 |
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| 作者姓名： | 张立营 张红 杨姝 吕茂民 尹惠琼 张改平 李德雪 章金刚 |
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| 作者单位： | 1. 军事医学科学院,野战输血研究所,国家生物医学分析中心,病毒安全检测实验室,北京,100850
2. 河南农业科学院,动物免疫学重点实验室,河南,郑州,450002 |
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| 基金项目： | 国家高技术研究发展计划(863计划) |
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| 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。
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| 关 键 词： | 蓝舌病毒 生物条形码检测 VP7蛋白 |
Bio-Bar Codes Assay for the Ultrasensitive Detection of Bluetonge Virus VP7 Protein |
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| ZHANG Li-Ying,ZHANG Hong,YANG Shu,LV Mao-Min,YIN Hui-Qiong,ZHANG Gai-Ping,LI De-Xue,ZHANG Jin-Gan. Bio-Bar Codes Assay for the Ultrasensitive Detection of Bluetonge Virus VP7 Protein[J]. Letters in Biotechnology, 2008, 19(5): 697-700 |
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| Authors: | ZHANG Li-Ying ZHANG Hong YANG Shu LV Mao-Min YIN Hui-Qiong ZHANG Gai-Ping LI De-Xue ZHANG Jin-Gan |
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| Affiliation: | ZHANG Li-Ying, ZHANG Hong, YANG Shu, LU Mao-Min, YIN Hui-Qiong, ZHANG Gai-Pin, LI De-Xue, ZHANG Jin-Gang (1. Laboratory for Viral Safety of National Centre of Biomedical Analysis, Institute of Transfusion Medicine, Academy of Military Medical Sciences, Beijing 100850; 2. Henan Key Laboratory of Animal Immunology, Zhengzhou 450002; China) |
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| Abstract: | 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. |
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| Keywords: | bluetongue virus bio-bar codes assay VF7 protein |
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