RPA primer and probe for detecting oil fish and detection method

Abstract

The invention provides a set of RPA primers and probes for detecting oil fish, and the set of primers and probes is designed based on Cytb gene of the oil fish. The forward primer, the reverse primerand the probe are as follows: (1) an upstream primer SEQ ID No.1: 5 '-CCCCCTCCAATATCTCAGTATGGTGAAATTTTGGG-3' (2) a downstream primer SEQ ID No.2: 5'-GACAGATGTGGGCTACGGAGGCAAAGGCTGATCCT- 3', and (3) the probe SEQ ID No.3: 5 '-GGTGAAATTTTGGGTCCCTACTAGGCCTCTGCCTAATTTCCCAAATCCTTAC-3'. Compared with the prior art, the detection method disclosed by the invention has the advantages that the detection iscompleted within 15 to 20 minutes; according to the method, whether the oil fish exists in the sample or not can be rapidly detected, so that production enterprises can monitor and detect incoming materials through the method, the benefits of the enterprises are maintained, the food safety risk of the enterprises is reduced, and the loss of the benefits of the enterprises is reduced. Meanwhile, on-site law enforcement rapid detection is facilitated, and technical support is provided for government supervision.

Description

technical field [0001] The invention belongs to the fields of biosafety technology and molecular biology, and in particular relates to an RPA primer, a probe and a detection method for detecting oily fish. Background technique [0002] Oily fish contains a lot of wax that is difficult for the human body to digest, and it is easy to cause diarrhea symptoms, especially for infants and young children, which poses a greater food safety risk. Foreign countries specifically named the disease "Keriorrhea". Oil fish, commonly known as "round cod", "wax oil fish", "dragon cod", "water cod", "white jade dolphin". Taxonomically, it belongs to the genus Scombro in the order Perciformes, including Lepidocybium flavobrunneum and Ruvettus pretiosus. [0003] Oily fish is a large deep-sea fish. When selling, the head and tail are generally removed, and sold in small packages such as fish pieces and steaks. After cutting, oily fish is almost exactly the same as cod and squid, so it cannot b...

AI Technical Summary

Problems solved by technology

However, these methods need to rely on expensive precision equipment such as sequencers, fluorescent PCR instruments, etc., have high requirements for personnel operations and environmental conditions, and have high costs for equipment calibration and maintenance, so they are not suitable for mobile laboratories and on-site applications. Operational requirements for scenarios such as rapid inspection, factory purchase acceptance, and enterprise self-inspection

In addition, the Sanger sequencing method is limited by the detection principle (requires a single peak for each base to identify the base ATGC), and cannot detect multi-source samples, so it cannot be applied to the detection of adulterated and mixed samples

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