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目的建立适合国境口岸入境生物材料中快速判断是否含有人源基因的实时荧光PCR检测方法并应用于口岸出入境生物材料的检测,为入境生物材料监管提供技术支撑。方法根据人线粒体ND2基因核酸序列,设计并筛选适合实时荧光PCR检测的引物和探针,优化反应体系,评估方法的灵敏度、特异性、稳定性,建立快速、特异的人源基因荧光PCR检测方法。结果建立的检测方法灵敏度高、特异性强、稳定性好,可在2 h内完成入境生物材料的检测,并有效区分非人灵长类动物及常见哺乳动物基因。结论本方法适用于对入境生物材料中人源基因的快速、准确检测,为口岸科学执法提供依据。
Objective To establish a real-time fluorescence PCR detection method for rapid determination of human gene in immigration biological material at border crossings and apply it to the detection of immigration biological materials at ports to provide technical support for inbound biomaterials regulation. Methods According to the nucleotide sequence of human mitochondrial ND2 gene, primers and probes suitable for real-time PCR detection were designed and screened. The reaction system was optimized and the sensitivity, specificity and stability of the method were evaluated. A rapid and specific fluorescent PCR method was established . Results The detection method established has high sensitivity, strong specificity and good stability. It can detect inbound biological material within 2 h and effectively distinguish non-human primate and common mammalian genes. Conclusion This method is suitable for the rapid and accurate detection of human genes in immigrant biological materials and provides the basis for scientific enforcement at ports.