Method for quickly, qualitatively and quantitatively measuring Lactobacillus plantarum in probiotic dairy products

A Lactobacillus plantarum, qualitative assay technology, applied in microorganism-based methods, microbial assay/inspection, biochemical equipment and methods, etc. limited issues

Inactive Publication Date: 2010-05-26
INNER MONGOLIA AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional methods are easily affected by factors such as culture conditions and strain characteristics, and the detection efficiency is limited. Moreover, they are gre

Method used

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  • Method for quickly, qualitatively and quantitatively measuring Lactobacillus plantarum in probiotic dairy products
  • Method for quickly, qualitatively and quantitatively measuring Lactobacillus plantarum in probiotic dairy products
  • Method for quickly, qualitatively and quantitatively measuring Lactobacillus plantarum in probiotic dairy products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0105] Rapid qualitative detection of Lactobacillus plantarum in probiotic dairy products:

[0106] In this example, the plasmid pPT containing the target amplification fragment was used as a positive control; dH 2 O was used as a negative control; in addition, L.plantarum 1.2437, L.casei ATCC393, L.rhamnosus1.2134, L.acidophilus ATCC 4356, L.fermentum 1.1880, and traditional fermented milk yogurt samples 25# and 36# collected from Tibet were used as sample, see figure 2 .

[0107] Described L.rhamnosus 1.2134, L.plantarum 1.2437, L.coryniformis subsp.coryniformis 1.1879 were purchased from the China General Microorganism Culture Collection; L.acidophilus ATCC4356, L.casei ATCC393 were purchased from American Type Culture Collection. Bifidobacterium animalis Subsp.Lactis V9 and L.reuteri ZL 12-1-1 are preserved by the Ministry of Education Key Laboratory of "Dairy Biotechnology and Engineering" of Inner Mongolia Agricultural University.

[0108] a. Preparation of template ...

Embodiment 2

[0123] Quantitative determination of Lactobacillus plantarum in probiotic dairy products:

[0124] Quantitative determination of soymilk fermented by L. plantarum IMAU10156 preserved in our laboratory. The measurement steps are as follows:

[0125] a. Preparation of sample total RNA

[0126] Extraction by Trizol method: After grinding 500 mg of the sample in a mortar filled with liquid nitrogen, quickly add 1 mL of Trizol reagent, extract sample RNA according to the instructions of the Trizol kit, and then treat it with DNaseI twice to ensure that the DNA contamination in the RNA is completely eliminated. The purified RNA sample was obtained, the concentration was determined according to the method described in the specification of this application, and the RNA integrity was subjected to 1% agarose gel electrophoresis.

[0127] b. Reverse transcription amplification

[0128] Reaction system 10.0 μL: 2.0 μL 5× PrimerScript Buffer (TakaRa DRR063A), 0.5 μL PrimerScript RT Enzy...

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Abstract

The invention relates to a method for quickly, qualitatively and quantitatively measuring Lactobacillus plantarum in probiotic dairy products. The method comprises the following steps: aiming at the lactobacillus plantarum in probiotic dairy products, independently designing a species specificity primer of the 16S rRNA gene sequence of the lactobacillus plantarum, carrying out species specificity PCR and real-time fluorescent quantitative PCR reaction by using the primer, and analyzing through an agarose gel electrophoresis pattern and a real-time fluorescent quantitative PCR pattern, thereby establishing the method for simply, conveniently, quickly, accurately, qualitatively and quantitatively measuring lactobacillus plantarum in probiotic dairy products.

Description

【Technical field】 [0001] The present invention relates to the technical field of dairy products containing probiotics. More specifically, the present invention relates to a rapid qualitative and quantitative determination method of Lactobacillus plantarum in probiotic dairy products. 【Background technique】 [0002] Prebiotic dairy products have become a hot spot of people's attention because of their rich nutritional value and special probiotic effects. At present, there are many kinds of probiotics and dairy products containing probiotics on the market, and they are increasing at a high rate every year. The supervision and certification of its quality, especially in terms of qualitative and quantitative testing, lacks scientific, reasonable and rapid methods. The qualitative and quantitative detection of probiotics in traditional methods still adopts the methods of physiological and biochemical reactions and pure culture isolation and counting in selective medium. The tr...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N21/64C12R1/25
Inventor 张和平包秋华刘文俊
Owner INNER MONGOLIA AGRICULTURAL UNIVERSITY
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