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Development of duplex real-time polymerase chain reaction for simultaneous detection of oilfish- and escolar-derived components

文献类型： 外文期刊

作者： Yao, Lin 1 ; Xin, Hongmei 1 ; Qu, Meng 1 ; Jiang, Yanhua 1 ; Guo, Yingying 1 ; Li, Fengling 1 ; Li, Na 1 ; Tan, Zhijun 1 ; W 1 ;

作者机构： 1.Minist Agr & Rural Affairs, Key Lab Testing & Evaluat Aquat Prod Safety & Qua, 106 Nanjing Rd, Qingdao 266071, Peoples R China

2.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Qingdao, Peoples R China

关键词： oilfish; escolar; duplex real-time polymerase chain reaction; 16S rRNA gene

期刊名称：JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE （ 影响因子：3.638； 五年影响因子：3.802 ）

ISSN： 0022-5142

年卷期：

页码：

收录情况： SCI

摘要： BACKGROUND Oilfish (Ruvettus pretiosus) and escolar (Lepidocybium flavobrunneum) are often used to adulterate high-value aquatic products, causing serious economic losses to consumers, and even keriorrhea in severe cases. It was difficult to identify them by morphological features for these two fish were processed into steak or fillet. The purpose of this study, therefore, is to develop an accurate and efficient method for detecting the oilfish- and escolar-derived components. RESULTS By comparing the mitochondrial 16S ribosomal RNA gene sequences of oilfish, escolar, and 16 varieties susceptible to adulteration, specific primers/probes were designed, and a duplex real-time polymerase chain reaction (PCR) method was established to detect oilfish- and escolar-derived components. The specificity and sensitivity of the method were analyzed, and the method was used to analyze 70 commercial samples to evalutate its applicability to actual samples in the market. The method developed was highly specific, without any cross-reaction on the other 16 species, with a limit of detection (LOD) for DNA of 0.0002 ng mu L(-1)forR. pretiosusand 0.002 ng mu L(-1)forL. flavobrunneum. In addition, the LOD for mixed muscle tissues was 0.1 g kg(-1). Oilfish- and escolar-derived components were detected in 12 of the 70 commercial samples, a result that is consistent with the classic DNA barcoding test results. CONCLUSION The duplex real-time PCR method developed can be used to detect oilfish and escolar specifically, sensitively and accurately; it provides a good technical support for the identification of authentic aquatic products. (c) 2020 Society of Chemical Industry

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