Primer combination and application thereof in detection of lactobacillus paracasei in dairy products

A primer combination, dairy product technology, applied in the field of bioengineering, to achieve the effect of shortening the detection time

Pending Publication Date: 2022-02-01
JUNLEBAO DAIRY GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the rapid and accurate detection of Lactobacillus paracasei

Method used

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  • Primer combination and application thereof in detection of lactobacillus paracasei in dairy products
  • Primer combination and application thereof in detection of lactobacillus paracasei in dairy products
  • Primer combination and application thereof in detection of lactobacillus paracasei in dairy products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 A primer for detecting a via cheese lactoforma in a dairy product

[0053] (1) Primitive design principle

[0054] Using system development trees to homologously cluster analysis of amplification sequences, the GYRB gene amplification region of Lactobacillus CGMCC4691 is listed as a target, and system development trees are constructed; using ClustAlX1.81 to genbank lactic acid bacteria and its nearby The GyRB sequence is compared, and 30 sequences are selected;

[0055]After the above 30 sequences were compared with DAMBE 4.2.13 software, the same sequence was removed, and the system development trees were constructed using the 18 sequences retained;

[0056] Based on the GYRB gene, the system development tree of L.Paracasei is built using the neighbor-joining method. figure 1 As shown, this tree is divided into four main branches: branch I, branch II, branch III and branch IV; branch i contains only a species, L.PaCTobacillus Paracasei; branch II consists of two ...

Embodiment 2

[0066] Example 2 Determination of auxiliary queen cheese emulsion heat

[0067] This example is a determination experiment of auxiliary cleaner heat tranout time, and the specific experimental step is:

[0068] A 1 mL of the sub-cheese lactofachalona was taken in 8 centrifugal tubes, and the centrifuge X1-X8 was removed, and the cured tube X1-X8 was treated with a water bath for 0min, 1 min, 2 min, 4 min, 6 min, 7 min, respectively. After 8 min or 9min, immediately cooled, and the number of sub-cheese lactobacillus in each dispersion was measured by a flat plate count method;

[0069] The results showed that the number of centrifugal tubes X1-X8 showed that the number of live bacteria was: 4.5 × 10, respectively. 8 CFU / ml, 2.3 × 10 6 CFU / ml, 1.2 × 10 6 CFU / ml, 4.8 × 10 3 CFU / ml, 1.7 × 10 2 CFU / mL, 6CFU / mL, 0CFU / mL, 0CFU / mL, which are known, heat treatment 8 min lactophilica all fire extinguishes, so the 90 ° C water bath is selected for 8 min to perform inactivatio...

Embodiment 3

[0070] Example 3 Optimization Experiment of PMA processing conditions

[0071] This example is an optimized experiment of PMA processing conditions, in order to obtain optimum PMA concentration, exposure time exposure time to the exposure time to eliminate the DNA amplification of the dead strain, according to Table 1 Design three factors three levels of orthogonal test, According to the aseptic water was washed twice with the aseptic water after treatment, the genomic DNA was subjected to 100 L of sterile water, and genomic DNA was extracted with a boiling water method, and the genomic DNA extracted by QLAMP was used, according to PMA pairs The inhibition rate of cheese DNA determines the best conditions for PMA treatment:

[0072] Table 1

[0073]

[0074] The specific experimental steps are:

[0075] The vicequarter CGMCC4691 was cultured in MRS medium at 37 ° C for 24 h, and the cells were rinsed with sterile saline, diluted to 10 5 CFU / ml, the bacterial suspension is name...

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Abstract

The invention discloses a primer combination and an application thereof in detection of lactobacillus paracasei in dairy products, wherein the primer combination is based on a gyrB gene, and the specific primer combination is designed by establishing a phylogenetic tree of lactobacillus paracasei and analyzing strain characteristics of lactobacillus paracasei; and the designed primer combination is applied to a real-time fluorescent quantitative loop-mediated isothermal amplification (qLAMP) technology, so that the lactobacillus paracasei in the dairy products is detected. The primer combination is high in specificity and sensitivity, and lactobacillus paracasei in the dairy products can be rapidly detected. The primer combination is suitable for qualitative and quantitative detection of lactobacillus paracasei in the dairy products.

Description

Technical field [0001] The present invention belongs to the field of bioengineering, involving primers and methods of detection bacteria, specifically, the primers and corresponding detection methods of detecting parstrutinia. Background technique [0002] L. Paracasei, belongs to the thick-wallet door, bacillia, lactate, lactate, lactate species, belonging to Gram-positive bacteria, often used in milk and dairy products or auxiliary screwdriver, A probiotic, lactic acid bacteria, lactic acid bacteria, lactic acid bacteria in recent years, after the gastrointestinal tract digestion, the sub-cheese Lactobacillus can still survive in the intestine, and play a probiotics that promotes human digestion and absorption, regulates immunization, and improve intestinal microorological balance. effect. [0003] The tablet counting method is a traditional colonies, which is subjective, cumbersome, complicated, time-consuming, error, and unable to survive but not cultivated bacterial cells, a...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/6851C12N15/11C12R1/225
CPCC12Q1/689C12Q1/6851C12Q2531/119C12Q2563/107C12Q2531/113
Inventor 王世杰胡连霞杨婉秋袁庆彬薛玉玲冯丽莉荀一萍张栋封肖颖武明月
Owner JUNLEBAO DAIRY GRP CO LTD
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