不同食品加工方式对提取鮟鱇鱼肉DNA的影响

Effects of different food processing methodson DNA quality of Lophius lituion

中文关键词:  食品加工  鮟鱇鱼  DNA提取  实时荧光PCR

英文关键词:food processing  Lophius lituion  DNA extraction  real-time PCR

中文摘要:

      目的 研究不同食品加工方式对提取鮟鱇鱼肉DNA的影响。方法 以实时荧光PCR方法检测鮟鱇鱼成分标准为例, 采取水煮、微波、油炸3种处理方式, 选用试剂盒提取DNA, 利用核酸蛋白测定仪测定其在260 nm、280 nm处的吸光度, 通过A260来计算核酸浓度, A260/A280来评估核酸的纯度。以提取的DNA为模板, 进行实时荧光PCR扩增。通过Ct值判断不同加工方式对实时荧光PCR结果的影响。结果 水煮、微波、油炸3种加工方式中, 油炸对DNA的影响最明显, 当在高温下油炸时间过久会导致鱼肉焦糖化反应及蛋白质变性严重, 变成具有一定孔隙的焦状物, 此时鱼肉在提取DNA的过程中难以消化, 造成DNA提取量少, 最终导致实时荧光PCR结果假阴性。而水煮、微波两种加工方式即使处理时间过久也没有对实时荧光PCR结果产生明显影响。结论 在鱼制品使用实时荧光PCR方法检测时, 应充分考虑加工方式对鱼制品DNA的影响。

英文摘要:

      Objective To investigate the impact of several common food processing methodson DNA quality ofLophius lituion. Methods The standard of real-time PCR method was used to detect DNA quality of Lophius lituion. Lophius lituion was processed by 3 different processing ways(water-boiling, microwave, frying). DNA was extracted by commercial DNA extractionkits.The purity of the DNA obtained from the samples was calculated according to A260/A280 values and the concentration of DNA was calculated by A260. The extracted DNA was amplified by real-time PCR. The influence of different processing methods on real-time PCR results was judged by Ct value. Results Among the 3 kinds of processing methods, frying revealed the most damaging effect on DNA. Long-time frying wouldlead to the occurrence ofserious caramelization and protein denaturation.Fish was turned into a porous cokeand difficult to extract DNA, whichledto less extraction of DNA and false negative results in real-time PCR.However, boiling and microwave processinghad no significant impact on the results of real-time PCR, even if the processing time wastoo long. Conclusion The effects of processing methods on DNA of fish products should be fully considered when using real-time PCR method for detecting fish products.