Quick qualitative and quantitative measuring method of fermented lactobacillus in probiotic milk products

A Lactobacillus fermentum, qualitative assay technology, applied in the directions of microorganism-based methods, microorganism determination/inspection, biochemical equipment and methods, etc., can solve problems such as large impact, limited detection efficiency, time-consuming and labor-intensive, etc.

Inactive Publication Date: 2010-02-17
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 greatly affected by external environmental factors and medium performance. The detection results lack stability and reliability, and are time-consuming and labor-intensive.

Method used

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  • Quick qualitative and quantitative measuring method of fermented lactobacillus in probiotic milk products
  • Quick qualitative and quantitative measuring method of fermented lactobacillus in probiotic milk products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0104] Rapid Qualitative Detection of Lactobacillus fermentum in Probiotic Dairy Products:

[0105] This embodiment uses the plasmid pFT containing the target amplified fragment as a positive control; dH 2 O was used as a negative control; in addition, L.fermentum 1.1880, L.fermentum 18302 (IMAU60070), traditional fermented lactic yogurt sample 25# collected from Tibet, natural fermented lactic yogurt sample 36# collected from Tibet, L.casei ATCC393, L. .rhamnosus 1.2134, L. acidophilus ATCC 4356 and L. plantarum 1.2437 were carried out as samples.

[0106] The L.rhamnosus 1.2134 and L.plantarum 1.2437 were purchased from the China General Microorganism Culture Collection Center; L.acidophilus ATCC 4356 and L.casei ATCC393 were purchased from the American Type Culture Collection; It was isolated from yogurt by the Ministry of Education Key Laboratory of Biotechnology and Engineering, and the preservation number is IMAU60070.

[0107] a. Preparation of template DNA

[0108] ...

Embodiment 2

[0122] Quantitative determination of Lactobacillus fermentum in probiotic dairy products:

[0123] Quantitative determination of Lactobacillus fermentum was carried out on different samples mentioned in Example 1. Its determination steps are as follows:

[0124] a. Preparation of sample total RNA

[0125] 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 integrity of the RNA was electrophoresed through 1% agarose gel.

[0126] b. Reverse transcription amplification

[0127] Reaction system 10.0 μL: 2.0 μL 5×PrimerScript Buff...

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Abstract

The invention relates to a quick qualitative and quantitative measuring method of fermented lactobacillus in probiotic milk products. Aiming at fermented lactobacillus in probiotic milk products, themethod designs a species specific primer of a 16S rRNA gene sequence of fermented lactobacillus by self, the species specific primer PCR reaction and real-time fluorescent quantitative PCR reaction ofthe primer is carried out to establish the simple, convenient, quick and accurate qualitative and quantitative measuring method of fermented lactobacillus in probiotic milk products by agarose gel electrophoresis pattern analysis and real-time fluorescent quantitative PCR pattern analysis.

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 fermentum 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 affec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/64C12R1/225
Inventor 张和平包秋华张家超
Owner INNER MONGOLIA AGRICULTURAL UNIVERSITY
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