食源性致病菌核酸高效提取方法

High-efficiency extracting method of food-borne pathogen nucleic acid

Abstract

食源性致病菌核酸高效提取方法,PCR技术目前已被广泛用于病原微生物的检测。PCR检测技术的第一步就是模板核酸的制备,即样本中核酸的提取,这直接影响着PCR反应的结果。长期以来,样本中核酸的提取和纯化一直是耗时、繁琐的过程,严重减慢了检测速度。因此,许多学者一直在探索各种核酸的提取方法。本发明其组成包括:将细菌培养液离心,向离心沉淀中加入缓冲液A,震荡并加入溶菌酶,温育后加入蛋白酶K,加入缓冲液B,剧烈振荡后,在温度65℃温育10分钟,制成裂解溶液,通过硅基质SiO2材料的吸附膜收集高质量的核酸提取核酸溶液。本发明应用于病原微生物的检测。
The present invention provides a high-efficiency extracting method of food-borne pathogen nucleic acid. A PCR technology has been widely used for the detection of pathogenic microorganisms currently. A first step of the PCR detecting technology is the preparation of template nucleic acid, that is, the extraction of nucleic acid in a sample, which directly affects the result of PCR reaction. For along time, the processes of extraction and purification of the nucleic acid in a sample consume time, and are fussy all the time, so detection speed is seriously slowed down. Thus, a plurality of scholars searches after the extracting method of various nucleic acids. The high-efficiency extracting method of food-borne pathogen nucleic acid comprises the following steps: bacterial culture solution is centrifuged; buffer solution A is added into centrifugal sediment to be vibrated, and muramidase is added; prolease K is added after incubation; buffer solution B is added to be incubated at the temperature of 65 DEG C for 10 minutes after being vibrated violently to be prepared into cracking solution; and high-quality nucleic acid is collected by an adsorption film of silicon matrix SiO2 material is used for extracting nucleic acid solution. The present invention is applied to the detection of the pathogenic microorganisms.

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Patent Citations (1)

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