Rapid nano PCR detection method for heat-proof acid-resistant bacterium in concentrated fruit juice

A technology of concentrated fruit juice and detection method, which is applied in the field of detection of heat-resistant acidophilus in concentrated fruit juice, can solve the problems of concentrated fruit juice safety and quality control, restrict the export of concentrated fruit juice, and cannot give timely feedback, etc., and achieve huge direct economic benefits and improve Juice quality and safety level, time-saving effect

Inactive Publication Date: 2012-03-28
TIANJIN UNIV OF SCI & TECH
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Problems solved by technology

These traditional culture counting detection methods take a long time, and generally take 3 days or even a week to produce a test report, which cannot be fed back to the production line in time, thus becoming a difficult problem in the safety and quality control of concentrated fruit juices, which seriously restricts the export of concentrated fruit juices in my country

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  • Rapid nano PCR detection method for heat-proof acid-resistant bacterium in concentrated fruit juice
  • Rapid nano PCR detection method for heat-proof acid-resistant bacterium in concentrated fruit juice
  • Rapid nano PCR detection method for heat-proof acid-resistant bacterium in concentrated fruit juice

Examples

Experimental program
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Effect test

Embodiment 1

[0070] Rapid PCR Detection of Heat-resistant and Acid-resistant Bacteria in Concentrated Apple Juice

[0071] 1. Isolation of thermotolerant acidophilus from concentrated apple juice:

[0072] Take 100ml of concentrated apple juice and dilute it 10 times with sterile water, heat shock the diluted juice in a water bath at 80°C for 10 minutes, then cool it to room temperature with cold water, and filter it with a 0.45 μm sterile filter membrane. After the filter membrane, use sterile medical tweezers Take it off and put it into a small Erlenmeyer flask containing 25ml of liquid culture medium and cultivate it on a shaker for 6 hours before use.

[0073] The preparation method of the liquid medium is (weight ratio): 0.25% of yeast extract powder, 0.5% of peptone, 0.1% of glucose, and 0.1% of Tween-80 are adjusted to a pH value of 3.7 with 250g / L sterile malic acid, and the The temperature is 43°C and the rotating speed of the shaker is 120 rpm.

[0074] 2. Extraction of the who...

Embodiment 2

[0088] 1. Isolation of thermotolerant acidophilus from concentrated orange juice:

[0089] Take 100ml of concentrated orange juice and dilute it 10 times with sterile water, heat shock the diluted juice in a water bath at 80°C for 10 minutes, then cool it to room temperature with cold water, and filter it with a 0.45 μm sterile filter membrane. After the filter membrane, use sterile medical forceps Take it off and put it into a small Erlenmeyer flask containing 25ml of culture solution, and cultivate it on a shaker for 6 hours before use. (The liquid medium is composed of yeast extract powder 0.25%, peptone 0.5%, glucose 0.1%, Tween-800.1%, adjust its pH value to 3.7 with 250g / L sterile malic acid.)

[0090] 2. Extraction of the whole genome of thermostable acidophilus:

[0091] Use a 15ml conical centrifuge tube to centrifuge at 5000rpm for 10min, discard the supernatant, add 20μl of 5% Chelex100 to the bacterial pellet, vortex and oscillate to mix, bathe in water at 100°C f...

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Abstract

The invention discloses a rapid nano-PCR detecting method of thermotolerant acid-resistant bacteria, which comprises the following steps: (1)separating thermotolerant acidophilic bacteria from the concentrated juice; (2)extracting the whole genome from thermotolerant acidophilic bacteria; (3)allocating rapid PCR system depended on nanometer silver; (4)setting and operating the rapid PCR condition; (5)detecting the PCR product with total time at 7.5h. The invention can monitor the thermotolerant acid-resistant bacteria of the condensed juice in the industrializing manufacturing effectively, which enlarges the directly economic benefit of export trade.

Description

Technical field: [0001] The invention belongs to the technical field of molecular biology, in particular to a method for detecting heat-resistant acidophilus in concentrated fruit juice by using a rapid nano-PCR method. Background technique: [0002] my country is currently one of the largest producers of juice in the world, and concentrated juice is exported in large quantities every day. However, a new type of contaminating bacteria in fruit juice - thermostable acidophilus has been reported abroad. The scientific name of heat-resistant and acid-resistant bacteria is Alicyclobacillus acidoterrestris, which is a thermophilic, acidophilic and aerobic bacillus. Its spores survive pasteurization in juice processing. Due to the failure of pasteurization to remove this bacteria and insufficient washing of raw fruit, etc., heat-resistant and acid-resistant bacteria are carried into the final product in the production of concentrated juice. Although the bacterium does not multi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04
Inventor 张治洲沈涔超刘常金
Owner TIANJIN UNIV OF SCI & TECH
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