摘要
目的:建立一种快速、高通量的检测牙膏中7种致病微生物的基因膜芯片方法。方法:设计7种致病微生物的特异性探针固定在尼龙膜上,样品增菌培养后提取DNA,与多重PCR扩增产物杂交,通过酶-底物显色产生肉眼可见的斑点。结果:建立的方法特异性强,重复性好,准确度高,检出限为核酸0.01 ng,满足检测要求。结论:本研究建立了同时检测牙膏中7种致病微生物的基因膜芯片法,提高了检测效率,为牙膏的微生物检测方法提供了新思路。
Objective:To develop a fast and high-throughput method for simultaneous detection of seven pathogenic microorganisms in toothpastes employing multiplex PCR combined with gene membrane chips.Methods:Designed specific probes for seven pathogenic microorganisms in toothpastes were fixed on nylon membranes,hybridizes with multiplex PCR amplification products,and generated visible spots through enzyme substrate color development.Results:The method showed strong specificity,good repeatability and high accuracy,and demonstrated an absolute sensitivity of 0.01 ng in the multiplex PCR and membrane chip hybridization system,meeting the detection requirements.Conclusion:The developed method for simultaneous detection of seven pathogenic microorganisms in toothpastes,employing multiplex PCR combined with gene membrane chips,can enhance the detection efficiency and provide reference for molecular detection of microbials in toothpastes.
作者
刘艳
金鹏
王萍
杭婧
李鑫
邢海艳
LIU Yan;JIN Peng;WANG Ping;HANG Jing;LI Xin;XING Haiyan(Huai’an Institute for Food and Drug Control,Huai’an 223200,China)
出处
《药学与临床研究》
2025年第4期323-327,共5页
Pharmaceutical and Clinical Research
基金
江苏省市场监督管理局科技计划项目(KJ2024090、KJ2025082)
淮安市科学技术局淮安市基础研究计划(指导性)项目(HABZ202331)。
关键词
牙膏
致病微生物
基因膜芯片
多重聚合酶链式反应
Toothpaste
Pathogenic microorganisms
Gene membrane chip
Multiplex polymerase chain reaction
