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Primer set and application of real-time fluorescence quantitative PCR for detection of luxs gene of lactic acid bacteria

A real-time fluorescent quantitative, lactic acid bacteria technology, applied in the field of lactic acid bacteria LuxS gene detection, to achieve the effect of strong primer specificity

Active Publication Date: 2022-04-01
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Primers are one of the materials needed for real-time fluorescent quantitative PCR detection. After investigation, there are few reports on the primer sets of Lactobacillus bulgaricus, Streptococcus thermophilus and Lactobacillus acidophilus

Method used

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  • Primer set and application of real-time fluorescence quantitative PCR for detection of luxs gene of lactic acid bacteria
  • Primer set and application of real-time fluorescence quantitative PCR for detection of luxs gene of lactic acid bacteria
  • Primer set and application of real-time fluorescence quantitative PCR for detection of luxs gene of lactic acid bacteria

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Experimental program
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Embodiment Construction

[0023] Below in conjunction with embodiment the present invention is described in further detail.

[0024] 1. Selection of internal reference genes

[0025] Taking Lactobacillus acidophilus CICC6074 before and after gastrointestinal fluid treatment as the research object, dp3D (DNA polymerase Ⅲ, delta subunit), ldhD (L-lactate dehydrogenase), gapdH (Glyceraldehyde-3-phosphate dehydrogenase) and gryA (gryA CDS) expression stability was evaluated.

[0026] The gene sequences of dp3d, ldhD, gapdH and gryA were obtained from the NCBI database and Geneious software, and the corresponding primers were designed using primer premier 6 software, and the specificity of the primers was verified by BLAST analysis. The selection criteria followed when designing the primers are: the primer length is 18-25bp, the GC content is about 50%, and the Tm is about 55°C. The results of primer design are shown in Table 1.

[0027] Table 1

[0028]

[0029]

[0030] Lactobacillus acidophilus...

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Abstract

The invention relates to a primer set for detecting the LuxS gene of lactic acid bacteria by real-time fluorescent quantitative PCR, including a primer pair for detecting the internal reference gene of lactic acid bacteria and a primer pair for detecting the LuxS gene of lactic acid bacteria, and the lactic acid bacteria include Lactobacillus bulgaricus, Streptococcus thermophilus and acid Lactobacillus, the internal reference gene is dp3d. It can more accurately reflect the synthesis of AI-2 signal molecules. The invention also relates to the application of the primer set in the preparation of the real-time fluorescent quantitative PCR detection kit for the LuxS gene of lactic acid bacteria, which can more accurately reflect the synthesis of AI-2 signal molecules when the three bacteria are co-cultured.

Description

technical field [0001] The invention relates to the technical field of detection of lactic acid bacteria LuxS gene, in particular to a primer set for real-time fluorescent quantitative PCR detection of lactic acid bacteria LuxS gene and its application. Background technique [0002] Lactic acid bacteria are a ubiquitous physiological flora present in the human intestinal tract, which can colonize the human intestinal tract and form a protective film to protect bacteria from the harsh gastrointestinal tract (GIT) microenvironment. At the same time, short-chain fatty acids and lactic acid produced by lactic acid bacteria have been shown to play an important role in maintaining the balance of the intestinal environment. In addition, lactic acid bacteria can also improve the nutritional value of fermented foods, reduce intestinal infections and serum cholesterol levels. Lactic acid bacteria are more resistant to acids and bile salts in the gastrointestinal environment than othe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/6851C12Q1/14C12Q1/10C12N15/11
CPCC12Q1/689C12Q1/6851C12Q2600/166C12Q2531/113C12Q2563/107C12Q2545/101
Inventor 吴振余艺星温健竹潘道东夏强曾小群何俊
Owner NINGBO UNIV