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

A bacillus and detection method technology, applied in the field of PCR rapid and specific detection of Alicyclobacillus in apple juice, can solve the problems of expensive equipment and reagents, long detection time, etc., and achieve the effect of low cost

Inactive Publication Date: 2011-07-27
NORTHWEST A & F UNIV
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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 conve

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

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Embodiment Construction

[0021] The present invention uses microwaves to directly extract DNA from fruit juice samples, according to Alicyclobacillus acidoterrestris DSM 3922 provided in GenBank T A pair of specific primers were designed based on the squalene cyclase (Shc) gene sequence, and the primer specificity, microwave extraction DNA amplification effect and detection sensitivity were studied respectively. After verification, the detection time was shortened and the detection cost was reduced. It can realize the online detection of apple juice production.

[0022] Specifically follow the steps below:

[0023] 1) Take 1ml fruit juice bacteria suspension, centrifuge at 10000r / min for 2min, discard the supernatant, add 50ul double distilled water to resuspend;

[0024] 2) The resuspended cells were microwaved at 1000W for 60s, centrifuged at 5000r / min for 1min, and the supernatant was taken to obtain the template DNA;

[0025] 3) Design a pair of SHC primers JLX-F and JLX-R, where:

[0026] Prim...

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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...

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
Inventor 樊明涛焦凌霞
Owner NORTHWEST A & F UNIV
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