| 三种禽用疫苗外源病原多重荧光PCR检测方法建立 |
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| 作者姓名： | 王艳 张强 李健 张建峰 朱小清 |
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| 作者单位： | 上海海关;广东省农业科学院动物卫生研究所;北京兰伯瑞生物技术有限公司 |
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| 摘 要： | 根据GenBank中收录的鸡毒支原体、禽白血病病毒和网状内皮细胞增生症病毒的基因序列,分别设计并合成了引物和相应的荧光探针,通过对反应条件和反应程序的优化,建立了一种可以同时检测3种禽用疫苗外源性病原污染因子的三重荧光PCR检测方法。结果显示,建立的三重荧光PCR灵敏度高,最低可检测到10 copies/μL的质粒,并具有良好的特异性,可用于禽用疫苗外源性污染因子的快速检测,也可用于相关疫病的诊断。
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| 关 键 词： | 鸡毒支原体 禽白血病病毒 网状内皮细胞增生症病毒 实时荧光PCR |
Development of a Triplex Real-time PCR Assay for Detection of MG,ALV, REV in Poultry Vaccines |
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| Authors: | WANG Yan ZHANG Qiang LI Jian ZHANG Jian-Feng ZHU Xiao-Qing |
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| Institution: | (Shanghai Customs,Shanghai 200135;Institute of Animal Health,Guangdong Academy of Agricultural Science,Guangzhou 510640;Beiing Laboratory Biology Technology co.,Ltd,Beiing 101300) |
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| Abstract: | Based on the sequences of mycoplasma gallisepticum(MS), Avian Leukosis virus(ALV) and reticuloendotheliosis virus(REV) genome from GenBank, three sets of primers and probes for the three pathogens were designed and synthesized. After the reaction conditions and processes were optimized, a triplex real-time PCR assay was established. The results showed that this assay had reasonable sensitivity, and the detection limits for plasmids was 10 copies/μL without cross reaction to other pathogens. This realtime PCR assay reported in the study can be used for the diagnosis of related diseases and rapid detection of exogenous contamination of poultry vaccines. |
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| Keywords: | MG ALV REV real-time PCR |
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