Real-time PCR法检测水产品中异鳞蛇鲭、棘鳞蛇鲭源性成分

doi:10.7506/spkx1002-6630-20190729-394

食品科学 ›› 2020, Vol. 41 ›› Issue (24): 273-280.doi: 10.7506/spkx1002-6630-20190729-394

Real-time PCR法检测水产品中异鳞蛇鲭、棘鳞蛇鲭源性成分

信红梅，姚琳，陆键萍，曲梦，江艳华，李风铃，郭莹莹，王联珠

（1.大连工业大学食品学院，辽宁大连 116034；2.农业农村部水产品质量安全检测与评价重点实验室，中国水产科学研究院黄海水产研究所，山东青岛 266071；3.中国海洋大学食品科学与工程学院，山东青岛 266000）

出版日期:2020-12-25 发布日期:2020-12-28
基金资助:
“十三五”国家重点研发计划重点专项（2016YFF0201805）

Detection of Lepidocybium flavobrunneum- and Ruvettus pretiosus-Derived Components in Aquatic Products by Real-time PCR

XIN Hongmei, YAO Lin, LU Jianping, QU Meng, JIANG Yanhua, LI Fengling, GUO Yingying, WANG Lianzhu

(1. School of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, China; 2. Key Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; 3. College of Food Science and Engineering, Ocean University of China, Qingdao 266000, China)

Online:2020-12-25 Published:2020-12-28

摘要/Abstract

摘要： 目的：建立异鳞蛇鲭、棘鳞蛇鲭实时聚合酶链式反应（real-time polymerase chain reaction，real-time PCR）检测方法。方法：对比异鳞蛇鲭、棘鳞蛇鲭、大西洋鳕等18 种鳕鱼及易掺假品种的线粒体细胞色素C氧化酶I（cytochrome C oxidase subunit I，COI）基因，设计异鳞蛇鲭、棘鳞蛇鲭物种特异性引物/探针，建立两种鱼的real-time PCR检测方法，评估其特异性、绝对灵敏度和相对灵敏度；用该方法对50 份市售标称“鳕鱼”制品进行检测，评估其实际使用效果。结果：两种检测方法特异性强，绝对灵敏度为0.002 ng/μL（异鳞蛇鲭）和0.000 2 ng/μL（棘鳞蛇鲭），相对灵敏度均达0.01%，满足实际检测需求。在50 份市售样品中检出1 份含异鳞蛇鲭成分，1 份含棘鳞蛇鲭成分，与测序比对结果一致。结论：本研究建立的异鳞蛇鲭和棘鳞蛇鲭源性成分的检测方法特异性强、灵敏度高，为保护消费者权益和水产品掺杂使假的检测提供技术支持。

关键词: 异鳞蛇鲭；棘鳞蛇鲭；油鱼；实时聚合酶链式反应；COI基因

Abstract: Objective: To develop a real-time polymerase chain reaction (real-time PCR) method for the detection of Lepidocybium flavobrunneum- and Ruvettus pretiosus-derived components. Methods: The mitochondrial cytochrome C oxidase subunit I (COI) gene sequences of 18 cod species including L. flavobrunneum, R. pretiosus and Gadus morhua and other species commonly used as adulterants were aligned with one another, and L. flavobrunneum- and R. pretiosus-specific primers and probes were designed and used to establish a real-time PCR method for the detection of the two fishes. The specificity, absolute sensitivity and relative sensitivity of the PCR method were assessed. In order to evaluate the practicability, 50 commercial products labeled as cod were tested by this method. Results: This method showed high specificity, with absolute sensitivity of 0.002 and 0.000 2 ng/μL for L. flavobrunneum and R. pretiosus, respectively, and relative sensitivity of 0.01% for both so that it could be applied to detect actual samples. The two fishes were separately detected in one of the 50 samples, consistent with sequence alignment. Conclusion: The proposed method in this study displayed good specificity and sensitivity towards the detection of L. flavobrunneum- and R. pretiosus-derived components, and could provide technical support for the detection of adulterated aquatic products to protect consumer rights.

Key words: Lepidocybium flavobrunneum; Ruvettus pretiosus; oilfish; real-time polymerase chain reaction; COI gene

中图分类号:

TS254.7

信红梅，姚琳，陆键萍，曲梦，江艳华，李风铃，郭莹莹，王联珠. Real-time PCR法检测水产品中异鳞蛇鲭、棘鳞蛇鲭源性成分[J]. 食品科学, 2020, 41(24): 273-280.

XIN Hongmei, YAO Lin, LU Jianping, QU Meng, JIANG Yanhua, LI Fengling, GUO Yingying, WANG Lianzhu. Detection of Lepidocybium flavobrunneum- and Ruvettus pretiosus-Derived Components in Aquatic Products by Real-time PCR[J]. FOOD SCIENCE, 2020, 41(24): 273-280.

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Real-time PCR法检测水产品中异鳞蛇鲭、棘鳞蛇鲭源性成分

Detection of Lepidocybium flavobrunneum- and Ruvettus pretiosus-Derived Components in Aquatic Products by Real-time PCR

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