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Method for rapidly detecting kinds of lactobacillus in fermented dairy product

A technology for fermenting dairy products and lactic acid bacteria, applied in biochemical equipment and methods, and microbial measurement/inspection, etc., can solve the problems of lack of stability and reliability of test results, limited detection efficiency, time-consuming and labor-intensive, etc., and achieve perfection Effects of quality supervision, improvement of testing capacity, and simplification of procedures

Inactive Publication Date: 2010-06-02
INNER MONGOLIA AGRICULTURAL UNIVERSITY
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  • Application Information

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Problems solved by technology

This method is easily affected by factors such as culture conditions and strain characteristics, and the detection efficiency is limited. It is also greatly affected by external environmental factors and medium performance. The detection process is time-consuming and labor-intensive, and the detection results lack stability and reliability.

Method used

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  • Method for rapidly detecting kinds of lactobacillus in fermented dairy product
  • Method for rapidly detecting kinds of lactobacillus in fermented dairy product
  • Method for rapidly detecting kinds of lactobacillus in fermented dairy product

Examples

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Embodiment 1

[0037] The rapid detection of lactic acid bacteria species in embodiment 1 yoghurt

[0038] The rapid detection of lactic acid bacteria species in yoghurt comprises the following steps:

[0039] 1. Extraction of metagenomic DNA:

[0040] Using liquid nitrogen freeze-thaw-CTAB method: Take 1.0mL of the three samples of yoghurt and place them in 5.0mL centrifuge tubes, immediately place them in liquid nitrogen to freeze completely, take them out and put them in a 65°C water bath to thaw, repeat freeze-thaw 3 times , add 3.0mL DNA extraction solution (100mol / L Tris-HCl; 100mmol / L EDTA; 100mmol / L Na 3 PO 4, 1.5mol / L NaCl, 1% CTAB, pH8.0) mix well, add 0.3mL 10% SDS and 10.0μL 10mg / mL proteinase K, shake at 200r / min in a constant temperature shaker at 37℃ for 2h, centrifuge at 10303g at room temperature After 10 min, the supernatant was collected and transferred to another centrifuge tube. The supernatant and an equal volume of chloroform were centrifuged at 10303 g for 10 min,...

Embodiment 2

[0052] The rapid detection of lactic acid bacteria species in embodiment 2 lactic acid bacteria fermented beverage

[0053] The rapid detection steps of lactic acid bacteria in lactic acid bacteria fermented beverages are as follows:

[0054] 1. Extraction of metagenomic DNA:

[0055] Liquid nitrogen freeze-thaw-CTAB method: Take 1.0 g of the four samples of lactic acid bacteria fermented beverages and place them in 5.0 mL centrifuge tubes, immediately place them in liquid nitrogen to completely freeze, take them out and put them in a 65°C water bath to thaw, and freeze and thaw repeatedly for 3 Once, add 3.0mL DNA extraction solution (100mol / L Tris-HCl; 100mmol / L EDTA; 100mmol / L Na 3 PO 4 , 1.5mol / L NaCl, 1% CTAB, pH8.0) mix well, add 0.3mL 10% SDS and 10.0μL 10mg / mL proteinase K, shake at 200r / min in a constant temperature shaker at 37℃ for 2h, centrifuge at 10303g at room temperature After 10 min, the supernatant was collected and transferred to another centrifuge tube. ...

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Abstract

The invention relates to a method for rapidly detecting the kinds of lactobacillus in a fermented dairy product. In the method, a method combing a polymerase chain reaction technique with a denatured gradient gel electrophoresis technique is applied; the existence of lactobacillus strains is determined by amplifying a lactic acid 16S rDNA V3 variable region and analyzing a polyacrylamide gel electrophoresis pattern so as to rapidly finish the detection of the kinds of lactobacillus in the fermented dairy product in two workdays.

Description

【Technical field】 [0001] The invention belongs to the field of detection of beneficial microorganisms in food, and in particular relates to the rapid qualitative detection of lactic acid bacteria in fermented milk products by using a method combining PCR technology and denatured gradient gel electrophoresis (DGGE) technology. 【Background technique】 [0002] Fermented dairy products are increasingly sought after because of their unique taste and rich nutritional value. At present, there are many kinds of fermented dairy products in the market, good and bad, and each fermented dairy product claims to contain a variety of lactic acid bacteria. Therefore, the supervision and management of its quality, especially the qualitative detection of the types of lactic acid bacteria contained in it, is particularly important. The traditional methods for the qualitative detection of lactic acid bacteria in fermented dairy products still use the methods of physiological and biochemical rea...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
Inventor 张和平张家超刘文俊包秋华孙志宏
Owner INNER MONGOLIA AGRICULTURAL UNIVERSITY
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