加工方法对猪肉成分RQ-PCR检测结果影响的研究  

Influence of Different Processing Methods on Pork Relative Quantification by Real-time PCR

作　　者：杨艳[1] 王桂姬[1] 周广运[1] 任皓威[1] 夏秀芳[1] 刘宁[1] YANG Yan WANG Gui-ji ZHOU Guang-yun REN Hao-wei XIA Xiu-fang LIU Ning＊(College of Food Science, Northeast Agricultural University, Harbin 150030, Heilongjiang, China)

机构地区：[1]东北农业大学食品学院,黑龙江哈尔滨150030

出　　处：《食品研究与开发》2017年第1期147-151,共5页Food Research and Development

基　　金：黑龙江省应用技术与开发计划(重大项目)"调理肉制品加工关键技术及安全质量控制"(GA15B302)

摘　　要：为评价不同加工处理方法对肉制品猪肉成分实时定量PCR(RQ-PCR)检测结果的影响,研究建立混合肉制品标准品(猪肉质量百分比0.05%~20%),采用5种方法(煮制、微波加热、超声波、烤制、高温高压灭菌)对混合肉制品进行处理,提取肉制品总DNA,采用RQ-PCR扩增线粒体细胞色素b(Cytb),以真核生物18S r RNA为内参,通过△Ctt建立标准曲线并进行相对定量,比较△Ct之间的差异,并由此判断不同加工方法对相对定量的影响,结果显示,不同的加工处理方法均能引起DNA的降解。而通过RQ-PCR检测发现,虽然Ct值随DNA降解程度加深而升高,但△Ct变化较小。因此,△Ct法可作为分析肉制品中猪肉成分的可靠定量手段。To evaluate the influence of different processing methods on real-time PCR relative quantification,this study set up reference meat mixture（with pork adulteration from 0.05 % to 20 %）,five different methods（boiling,microwaving,sonication,baking,autoclaving）were applying to the reference meat mixture,then verified the total DNA degradation by gel electrophoresis. △Ct and the percentage of pork adulteration were used to establish standard curve,comparing the differences of △Cts to evaluate the influence of processing methods on relative quantification.The results showed that all the processing method could cause the degradation of DNA,among which autoclaving play the greatest role in DNA degradation,meanwhile the sonication was the slightest. In real-time PCR,Ct raising with processing continue but the changing of △Ct were small,△Ct can be used as a reliable analysis method for quantification pork in meat products.

关 键 词：实时荧光PCR 相对定量 猪肉 加工处理 影响 

分 类 号：TS251.7[轻工技术与工程—农产品加工及贮藏工程]