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

A technology for the qualitative determination of Lactobacillus casei, applied in the direction of microbial-based methods, microbial determination/inspection, biochemical equipment and methods, etc., can solve the lack of stability and reliability of test results, limited detection efficiency, and time-consuming Power and other issues

Inactive Publication Date: 2010-05-26
INNER MONGOLIA AGRICULTURAL UNIVERSITY
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  • Description
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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 casei in probiotic dairy products
  • Method for quickly, qualitatively and quantitatively measuring Lactobacillus casei in probiotic dairy products
  • Method for quickly, qualitatively and quantitatively measuring Lactobacillus casei in probiotic dairy products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0107] Rapid qualitative detection of Lactobacillus casei in probiotic dairy products:

[0108] This embodiment uses the plasmid pCT containing the target amplified fragment as a positive control; dH 2 O was used as a negative control; in addition, L.casei ATCC393, L.plantarum 1.2437, L.rhamnosus1.2134, L.acidophilus ATCC 4356, yogurt sample 12# (mixed fermentation of L.casei Zhang and Bifidobacterium animalis Subsp.Lactis V9), yogurt Sample 36# (traditional fermented milk collected from Tibet) was used as a sample.

[0109] The L.rhamnosus 1.2134 and L.plantarum 1.2437 were purchased from China General Microorganism Culture Collection Center; L.acidophilus ATCC 4356 and L.casei ATCC393 were purchased from American Type Culture Collection; L.casei Zhang and Bifidobacterium animalisSubsp.Lactis V9 Preserved by the Ministry of Education Key Laboratory of "Dairy Biotechnology and Engineering" of Inner Mongolia Agricultural University.

[0110] a. Preparation of template DNA

...

Embodiment 2

[0125] Quantitative determination of Lactobacillus casei in probiotic dairy products:

[0126] Quantitative determination of Lactobacillus casei was carried out on different samples mentioned in Example 1. Its determination steps are as follows:

[0127] a. Preparation of sample total RNA

[0128] Trizol method extraction: 500 mg of fermented milk product was ground in a mortar filled with liquid nitrogen, and 1 mL of Trizol reagent was added quickly, and the sample RNA was extracted according to the instructions of the Trizol kit, and then treated twice with DNaseI to ensure that the DNA in the RNA was completely eliminated Contamination, the purified RNA sample was obtained, and the concentration was determined according to the method described in the specification of this application, and the RNA integrity was detected by 1% agarose gel electrophoresis.

[0129] b. Reverse transcription amplification

[0130] Reaction system 10.0 μL: 2.0 μL 5×PrimerScript Buffer (TakaRa ...

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Abstract

The invention relates to a method for quickly, qualitatively and quantitatively measuring Lactobacillus casei in probiotic dairy products. The method comprises the following steps: aiming at the Lactobacillus casei in probiotic dairy products, independently designing a species specificity primer of the 16S rRNA gene sequence of the Lactobacillus casei, 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 casei in probiotic dairy products.

Description

【Technical field】 [0001] The invention relates to the technical field of dairy products containing probiotics. More specifically, the present invention relates to a method for rapid qualitative and quantitative determination of Lactobacillus casei in probiotic milk products. 【Background technique】 [0002] Probiotic dairy products have become a hot topic due to their rich nutritional value and special beneficial effects. At present, there are many kinds of probiotics and dairy products containing probiotics on the market, and they are growing at a high rate every year. The supervision and certification of its quality, especially in terms of qualitative and quantitative detection, lacks scientific, reasonable and rapid methods. The qualitative and quantitative detection of probiotics in the traditional method still adopts the methods of physiological and biochemical reactions and selective medium pure culture isolation and counting. Traditional methods are easily affected ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N21/64C12R1/245
Inventor 张和平包秋华张家超
Owner INNER MONGOLIA AGRICULTURAL UNIVERSITY
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