Real-time fluorescence PCR (polymerase chain reaction) reagent kit and real-time fluorescence PCR detection method for specific detection of salmonidae fishes

Abstract

The invention discloses a real-time fluorescence PCR (polymerase chain reaction) reagent kit and a real-time fluorescence PCR detection method for specific detection of salmonidae fishes. The real-time fluorescence PCR reagent kit comprises a PCR buffer solution, a dNTP (diethyl-nitrophenyl thiophosphate) and DNA (deoxyribonucleic acid) polymerase, an upstream primer shown in the SEQ (sequence) ID (identification code) NO.1 or NO.2, a downstream primer shown in the SEQ ID NO.3, and a fluorescent probe shown in the SEQ ID NO.4. The detection method includes: (1) extracting DNA of a sample to be detected; (2) subjecting the DNA of the sample and a negative reference sample to PCR reaction, and performing fluorescence detection; (3) selecting an FAM fluorescence detection mode, adjusting a base line to take 3-15 circulating fluorescence signals, setting a threshold value line to just exceed the highest point of a normal negative reference sample; and if a fluorescence increasing curve of the sample to be detected exceeds the threshold value line and increases in excellent logarithms, judging the sample to be positive. By the aid of the reagent kit and the method, ingredients of the salmonidae fishes can be detected and identified quickly and accurately, false positive rate of normal PCR amplification is reduced.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and nucleic acid detection, in particular to a real-time fluorescent PCR detection kit and detection method for specific detection and identification of salmonid fish in food, feed and other processed fish products. Background technique [0002] In recent years, food shoddy and counterfeit and shoddy incidents have occurred from time to time in our country and even in the world, and they are becoming more and more serious. Most salmonids are deep-sea migratory fishes. So far, scientists have discovered 66 species of salmonids in the world. Because the salmon meat, especially the "salmon" among them, contains high protein and OMEGA-3 fatty acids, the fat content is very low, and has high nutritional value and therapeutic effect, so it is loved by consumers. At present, the demand for salmonids in the domestic market is increasing, but its price is high. Driven by profit, salmonids in th...

AI Technical Summary

Problems solved by technology

[0004] In summary, in view of the fact that there is no specificity and high sensitivity detection method for salmonids in food, feed and fish processing products

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