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目的多基因表达分析系统(Gene expression Profiler genetic analysis system,Ge XP)是一种全新的基因表达分析平台,检测特异性强,检测速度和灵敏度明显提高。本研究利用Ge XP系统和多重PCR技术,结合我国口岸检疫查验主要呼吸道传染病现状,建立甲型流感病毒、乙型流感病毒、甲型H1N1病毒的检测技术方法。方法制备病毒标准品质粒,收集病原体样本,利用Gen Bank下载3种病毒的核苷酸序列,根据3种病毒的序列特征,利用Ge XP e Xpress Profiler设计特异性引物,优化条件使Ge XP系统能够特异性的检测对应病毒。结果优化后的引物能够检测3种病毒,产物经Ge XP检测,3个峰值位于166.04 bp、193.33 bp、251.42 bp,符合甲型流感病毒、乙型流感病毒、甲型H1N1病毒病原体质粒PCR产物条带大小,特异性强,无交叉反应。对已知临床样本的检测中,检出率高且特异性强。结论 Ge XP系统能够用于病原体高通量检测工作,有益于扩展检测途径,提高检测效率。
Objective Gene expression profiler genetic analysis system (Ge XP) is a new platform for gene expression analysis. It has high detection specificity and high detection speed and sensitivity. In this study, Ge XP system and multiplex PCR technology, combined with China’s quarantine inspection port quarantine of the status of major respiratory infectious diseases, the establishment of influenza A virus, influenza B, H1N1 virus detection technology methods. Methods The virus standard plasmids were prepared and the pathogen samples were collected. GenBank downloaded the nucleotide sequences of the three viruses. Based on the sequence characteristics of the three viruses, the specific primers were designed by using Ge XP e Xpress Profiler. The optimized conditions enabled the Ge XP system Specific detection of the corresponding virus. Results The optimized primers were able to detect three kinds of viruses. The products were detected by Ge XP. The three peaks were 166.04 bp, 193.33 bp and 251.42 bp, which were in line with the PCR products of Influenza A, B and H1N1 With the size, specificity, no cross-reaction. Detection of known clinical samples, the detection rate and specificity. Conclusion The Ge XP system can be used for pathogen high-throughput detection, which is beneficial to extend the detection pathways and improve the detection efficiency.