PCR技术在食品有害微生物检测中的应用.docVIP

PCR技术在食品有害微生物检测中的应用 摘 要：食品安全是一个越来越引起人们重视的问题，不仅影响到人们的生活、工作和健康，更是关系到国计民生的大事。目前我们国家虽然制定相关的食品病原菌检验方法，由于检测方法均采用经典、耗时较长的方法，且存在国内试剂供应不配套等问题，很难适应公共卫生事件应急处理快速反应的需要，随着时代的进步，检测方法的先进性、灵敏度、特异性要求都更高。分子生物学检测技术的发展，为微生物食物中毒快速检提供了良好的技术平台。本文详细介绍了PCR技术的原理特点和采用PCR技术检测食品有害微生物的操作流程，同时介绍了当前广泛应用的4种基于PCR技术基础上改进的PCR检测新技术，在此基础上提出了该技术的缺点和在应用过程中存在的问题，同时对其应用前景作了展望。 关键词： PCR检测；微生物；食品安全 Applications of PCR in the detection of food borne pathogens Abstract: The security of Food is an increasingly important issue people will not only affect peoples lives, work and health, it is also a major event related to the peoples livelihood. Although our country is currently formulating the relevant methods of food pathogen deteting, the classical of a longer time-consuming detection methods are used, reagents and supplies of the existence of domestic support and other issues, it is difficult to adapt to deal with public health emergencies need rapid response, with the the progress of the times, advanced detection methods, sensitivity and specificity requirements are higher. Detection of molecular biology technology for the rapid seizure of food poisoning micro-organisms to provide a good technology platform.This paper formulates the characteristics of PCR technique and the use of PCR detection of harmful microorganisms in food processes detailly, while the current widely used PCR technology based on four kinds of improvements based on PCR detection of new technologies, in this based on the technology in the application of the shortcomings and problems that exist in the process, while its application prospects are looked ahead. Keywords: PCR inspection； Microbiology； Food safety实时荧光定量PCR技术于1996年由美国Applied Biosystems公司推出，由于该技术不仅实现了PCR从定性到定量的飞跃，而且与常规PCR相比，它具有特异性更强、有效解决PCR污染问题、自动化程度高等特点，目前已得到广泛应用。所谓实时荧光定量PCR技术，是指在PCR反应体系中加入荧光基团，利用荧光信号积累实时监测整个PCR进程，最后通过标准曲线对未知模板进行定量分析的方法。 Ct 值的定义 在荧光定量PCR技术中，有一个很重要的概念Ct值。C代表Cycle，t代表threshold，Ct值的含义是：每个反应管内的荧光信号到达设定的域值时所经历的循环数（如图1所示）。荧光域值（threshold）的设定 PCR反应的前15个循环的荧光信号作为荧光本底信号，荧光