Method for quickly and specifically detecting Alicyclobacillus in apple juice through polymerase chain reaction (PCR)

Abstract

The invention discloses a method for quickly and specifically detecting Alicyclobacillus in apple juice through polymerase chain reaction (PCR). In the method, DNA is directly extracted from a juice sample by microwaves; a pair of specific primers are designed according to a gene sequence of squalene cyclase (Shc) of AlicyclobacillusacidoterrestrisDSM3922 provided by a GenBank; a 25ul system consisting of template DNA, a primer JLX-F and a primer JLX-R is used for PCR amplification; and a target strip is obtained at a position of 648bp through 1 percent agarose gel electrophoresis detection, and a detection result is obtained. The specificity of the primers, the amplification effect of microwave extracted DNA, and detection sensitivity are researched, and through inspection, the method has the advantages of short detection time and low detection cost and can realize online detection of apple juice production.

Description

technical field [0001] The invention relates to a detection method in the food industry, in particular to a rapid and specific PCR detection method for Alicyclobacillus in apple juice. Background technique [0002] Alicyclobacillus was first reported by Japanese scholars in 1967. This genus of bacteria is thermophilic and acid-resistant, and has unique viability. It can survive the pasteurization process under acidic conditions. After contaminating fruit juice and juice drinks, flat-cap rancidity occurs. There is no obvious change in the appearance of the fruit juice in the early stage of pollution. Once the conditions are suitable, the bacteria of this genus will grow and multiply rapidly, producing guaiacol and halophenols and other undesirable flavor substances, resulting in the loss of aroma, poor taste, and increased turbidity of the juice product. Precipitation is formed, and the quality of fruit juice deteriorates within the shelf life, which brings huge economic loss...

AI Technical Summary

Problems solved by technology

[0005] There are also domestic scholars who have established PCR and QC-PCR rapid detection methods for heat-resistant bacteria. The entire detection time is 4h to 5h, which is much shorter than the traditional time of 4d to 5d. However, since these methods cannot break through the conventional DNA extraction, a certain Therefore, the isolation and enrichment culture of the bacteria cannot be surpassed. The required detection time is still long, and the equipment and reagents used are expensive, which still has a certain gap with the application in production practice.

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