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目的建立一种SYBR Green荧光PCR法定性检测太平洋无须鳕鱼成分的分析SYBR Green荧光PCR定性检测方法。方法根据太平洋无须鳕鱼线粒体细胞色素氧化酶CoxⅠ基因中的保守基因序列设计一对太平洋无须鳕鱼特异性引物,通过SYBR Green荧光PCR法进行PCR扩增反应,可对DNA提取试剂盒提取的太平洋无须鳕鱼成分DNA进行特异性检测。结果该SYBR Green荧光PCR定性检测方法的引物对于太平洋无须鳕鱼成分的检测特异性良好;方法的灵敏度分别为0.06 ng/μL太平洋无须鳕鱼DNA和质量分数为0.01%太平洋无须鳕鱼肉粉。通过对市售太平洋无须鳕鱼片、太平洋鳕鱼片和太平洋无须鳕鱼肝油的检测,该方法可检测出鱼片中的太平洋无须鳕成分。结论该检测方法特异性强,灵敏度高,能够用于食品中太平洋无须鳕鱼成分的真实性鉴别。
Objective To establish a SYBR Green fluorescence PCR method for the qualitative detection of Pacific Cod ingredient analysis SYBR Green fluorescent PCR qualitative detection method. Methods A pair of pacific cod-free primers were designed according to the conserved gene sequences in Cox-I gene of Pacific Cod. The SYBR Green fluorescence PCR method was used to amplify the PCR products. The DNA extracted from the Pacific Cod Component DNA is specifically tested. Results The primers of the SYBR Green qualitative PCR assay showed good specificity for detection of Pacific Cod ingredients. The sensitivity of the method was 0.06 ng / μL Pacific cod-free DNA and 0.01% Pacific cod-free meat meal. This method detects Pacific hake ingredients in fish fillets through the testing of Cod Fillet, Pacific Cod Fillet and Pacific Cod Liver Oil in the commercial Pacific. Conclusion The method is highly specific and sensitive and can be used to authenticate cod-free ingredients in the Pacific Ocean.