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PCR detection method for detecting lactobacillus plantarum ST-III and primer and kit thereof

A technology of Lactobacillus plantarum and detection method, which is applied in the field of biological detection, to achieve the effect of shortening the experimental period, strong specificity and high sensitivity

Active Publication Date: 2013-10-02
BRIGHT DAIRY & FOOD CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to provide a fast, specific, high-sensitivity, low-cost molecular detection method for Lactobacillus plantarum ST-III strain in view of the current situation that there is no molecular detection method for Lactobacillus plantarum ST-III strain. Biological detection method

Method used

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  • PCR detection method for detecting lactobacillus plantarum ST-III and primer and kit thereof
  • PCR detection method for detecting lactobacillus plantarum ST-III and primer and kit thereof
  • PCR detection method for detecting lactobacillus plantarum ST-III and primer and kit thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The rapid qualitative identification of plantarum lactobacillus ST-III in the probiotic milk product of embodiment 1

[0035] There are a lot of fermented beverages on the market. As the owner of a series of products related to Lactobacillus plantarum ST-III, it is necessary to identify whether these fermented products contain Lactobacillus plantarum ST-III, which has a strong probiotic effect, so as to prevent counterfeiting. Protect the legitimate rights and interests of consumers.

[0036] a. Preparation of template DNA. The sample DNA was extracted by the liquid nitrogen freeze-thaw-CTAB method, and the specific steps were as follows:

[0037] (1) Take 1.0g probiotic fermented milk product or pure culture of ST-III in a 10.0mL centrifuge tube, add 3.0mL DNA extraction solution (100mmol / L Tris-HCl; 100mmol / L EDTA; 100mmol / L Na3PO4, 1.5mol / L NaCl, 1% CTAB, pH8.0) mixed evenly, immediately placed in liquid nitrogen to freeze completely, took it out and put it in a 65...

Embodiment 2

[0054] Example 2 specificity verification

[0055] Select 50 different strains of Lactobacillus plantarum and 100 strains of other species of Lactobacillus, use the primer sequence provided by the present invention to carry out PCR reaction, and then analyze the probiotics according to the agarose gel electrophoresis pattern, simply, quickly and accurately Whether dairy products contain Lactobacillus plantarum ST-III strain. See some results figure 1 . Experimental results show that the primers provided by the present invention can only amplify ST-III, but have no corresponding amplification for other bacterial strains. Specifically, the method includes the following steps:

[0056] 1. Genomic DNA Extraction

[0057] Liquid nitrogen freeze-thaw-CTAB method was used to extract the total DNA of probiotic samples and the total DNA of purely cultured lactic acid bacteria (Lactobacillus plantarum ST-III) in the sample, and the specific steps were as follows:

[0058] Take an a...

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Abstract

The invention discloses a PCR detection method for detecting lactobacillus plantarum ST-III and a primer and a kit thereof. The invention provides genetic marker molecules of the ST-III bacterial strain, which are the 242nd-867th continuous nucleotides of a sequence shown in the 1st-867th gene of KilA domain protein coding the lactobacillus plantarum ST-III. As a molecular detection method of lactobacillus plantarum ST-III, the invention can be used for rapidly detecting whether a lactobacillus plantarum ST-III bacterial strain exists in probiotic dairy products. The invention has the advantages of high sensitivity, high specificity. low detection cost, and the like, can conveniently distinguish lactobacillus plantarum ST-III bacterial strains from other different bacterial strains in lactobacillus plantarum of the same kind, and has very strong practical application value.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a PCR detection method for detecting the ST-III strain of Lactobacillus plantarum and its primers and kit. Background technique [0002] Probiotics refer to a single or specific mixture of microorganisms that can reach the gastrointestinal tract in the state of live bacteria when ingested in a certain amount, and can promote the health of humans or animals by regulating the composition of intestinal flora. In recent years, with the successive discoveries of its probiotic effects, the production and sales of probiotic products have also shown an upward trend year by year. However, the supervision and certification of the quality of probiotic products in the market currently lacks scientific, reasonable and rapid methods for qualitative and quantitative detection and identification, which leads to the abuse of strains and the phenomenon of misleading, making products...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/31C12N15/11C12Q1/68C12Q1/04
Inventor 陈臣任婧周方方艾连中
Owner BRIGHT DAIRY & FOOD CO LTD