3种常见呼吸道病毒检测基因芯片的制备

发布时间：2019-06-06 03:58

【摘要】：目的建立一种可同时检测鼻病毒、呼吸道合胞病毒以及腺病毒的基因芯片方法。方法根据GenBank已发表的鼻病毒、呼吸道合胞病毒和腺病毒的基因序列,应用生物学软件设计相应的引物和寡核苷酸探针,构建检测芯片;使用巢式PCR扩增标记目的片段,将其与检测基因芯片进行杂交和扫描分析;通过特异性、灵敏性和重复性试验评估基因芯片方法。结果设计的全部探针均能与相应的标记样品特异性杂交,其中探针1a和1b可特异检测鼻病毒;探针2a、2b和2c可特异检测呼吸道合胞病毒,探针3a和3b可特异检测腺病毒。基因芯片法检测3种病毒的最低拷贝数分别是2.59×102个/μl、2.21×101个/μl和2.05×102个/μl。结论构建的基因芯片可特异性检测样品中低滴度的鼻病毒、呼吸道合胞病毒以及腺病毒,有望为临床提供确诊依据。
[Abstract]:Objective to establish a gene chip method for simultaneous detection of rhinovirus, respiratory syncytial virus and adenoviruses. Methods according to the gene sequences of rhinovirus, respiratory syncytial virus and adenoviruses published by GenBank, the corresponding primers and oligodeoxynucleotides probes were designed by biological software, and the detection chip was constructed. The target fragment was amplified by nest PCR, crossed with the detection gene chip and analyzed by scanning, and the gene chip method was evaluated by specificity, sensitivity and reproducibility test. Results all the designed probes could specifically cross with the corresponding labeled samples, in which probes 1a and 1b could specifically detect rhinovirus, probe 2a, 2b and 2c could specifically detect respiratory syncytial virus, and probes 3a and 3b could specifically detect adenoviruses. The minimum copy number of the three viruses detected by gene chip method was 2.59 脳 102 / 渭 l, 2.21 脳 101 / 渭 l and 2.05 脳 102 / 渭 l, respectively. Conclusion the constructed gene chip can specifically detect low titer rhinovirus, respiratory syncytial virus and adenovirus in the sample, which is expected to provide diagnostic basis for clinical diagnosis.
【作者单位】：吉林大学畜牧兽医学院;军事医学科学院军事兽医研究所;吉林农业大学动物科技学院;长春中医药大学;
【基金】：吉林省科技支撑计划项目(No.20100416)
【分类号】：R373

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