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Primer probe, kit and method for detecting heterologous oil adulterated in deep-sea fish oil
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A technology of primer probe and deep-sea fish oil is applied in the field of biological detection to achieve the effects of short detection time, high specificity and accuracy, and less toxic and harmful substances.
Active Publication Date: 2022-02-22
南宁海关技术中心
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[0005] The purpose of the present invention is to provide a primer probe, kit and method for detecting heterogeneous oils in deep-sea fish oil, to solve the technical problems of existing deep-sea fish oil detection technology sensitivity and one-time identification of multiple mixed oils
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Embodiment 1
[0110] This example is to verify the specific detection of marine fish, salmon, and plant triple real-time fluorescent PCR amplification system in animal and vegetable oils
[0111] 1. Purchase 100mL of fat from chicken, duck, pig, cattle, sheep, grass carp (freshwater fish), silver carp (freshwater fish), and bighead carp (freshwater fish) from vegetable markets, supermarkets, and the Internet; salad oil 100mL; purchase about 230 capsules of 100mL genuine deep-sea fish oil with cod as raw material and about 200 genuine deep-sea fish oil with salmon as raw material 100mL from physical stores and online.
[0112] 2. Extract DNA from 11 samples according to the DNA extraction method of fish oil (which may contain other oils and esters) described in step 1, and store them at -20°C.
[0113] 3. The primer probe solution is the main component of the above kit, and 11 kinds of sample DNA are amplified with 18SrRNA and the multiple real-time fluorescent PCR detection system designed ...
Embodiment 2
[0120] This example is to verify the specific detection of marine fish, salmon, and plant triple real-time fluorescent PCR amplification system after salmon oil is mixed with vegetable oil
[0121] 1. Purchasing relevant oils and fats through the same purchasing channels as in Example 1.
[0122] Sample preparation 1-5: The salmon oil sample was mixed with soybean oil at volume ratios of 0.01%, 0.1%, 1%, 10%, and 100%, respectively, with a final volume of 100 mL.
[0123] Sample preparation 6-10: The salmon oil sample was mixed with rapeseed oil at volume ratios of 0.01%, 0.1%, 1%, 10%, and 100%, respectively, with a final volume of 100 mL.
[0124] Sample preparation 11-15: Salmon oil samples were mixed with palm oil at volume ratios of 0.01%, 0.1%, 1%, 10%, and 100%, respectively, with a final volume of 100 mL.
[0125] Sample preparation 16-20: Salmon oil samples were mixed with corn oil at volume ratios of 0.01%, 0.1%, 1%, 10%, and 100%, respectively, with a final volume of...
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Abstract
The invention discloses a primer probe for detecting heterogeneous oil mixed in deep-sea fish oil, including at least one, two or three of the following combined probes: primer probe for detecting marine fish components Group 1, including primer 1-2 and probe 1, wherein the nucleotide sequence of the primer 1-2 is shown in SEQ ID NO.1-2 respectively, and the nucleotide sequence of the probe 1 is as shown in SEQ ID NO.1-2 Shown in ID NO.3; The primer probe group 2 that is used to detect salmon composition comprises primer 4-5 and probe 2, wherein, the nucleotide sequence of described primer 4-5 is respectively as SEQ ID NO.4- 5, the nucleotide sequence of the probe 2 is shown in SEQ ID NO.6; the primer probe set 3 for detecting plant components includes primer 7-8 and probe 3, wherein the primer The nucleotide sequences of 7-8 are respectively shown in SEQ ID NO.7-8, and the nucleotide sequence of the probe 3 is shown in SEQ ID NO.9. The method of the invention can accurately, quickly and high-throughput carry out quality classification and adulteration identification of deep-sea fish oil.
Description
technical field [0001] The invention relates to the technical field of biological detection, in particular to a primer probe, a kit and a method for detecting heterologous oil mixed in deep-sea fish oil. Background technique [0002] Deep-sea fish oil generally refers to the unsaturated fatty acid Omega-3 extracted from Osteichthyes fish deep in the ocean, and its main components are EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid). Acid) is an indispensable and important nutrient for the human body. In recent years, deep-sea fish oil has sprung up in the health care product market. It is regarded as a must-have product for regulating blood lipids, softening blood vessels, and cleaning up vascular garbage. The annual sales volume is about 100 million bottles, and the annual growth rate is as high as 30%. One of the highest quality deep-sea fish oils is extracted from Atlantic salmon (salmon) from Northern Europe and the Atlantic coast of France. However, there ar...
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