Method and kit for quickly and quantificationally detecting lactobacillus plantarum ST-III

A technology for quantitative detection of Lactobacillus plantarum, applied in biochemical equipment and methods, microbial measurement/inspection, fluorescence/phosphorescence, etc., to achieve the effects of shortened experimental period, simple operation, and fast detection speed

Active Publication Date: 2012-05-16
BRIGHT DAIRY & FOOD
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Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a rapid, specific, high sensitivity, and low-cost method for quantitatively detecting Lactobacillus plantarum ST-III for the current situation that there is no molecular detection method for Lactobacillus plantarum ST-III strain. Method and its kit

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  • Method and kit for quickly and quantificationally detecting lactobacillus plantarum ST-III
  • Method and kit for quickly and quantificationally detecting lactobacillus plantarum ST-III
  • Method and kit for quickly and quantificationally detecting lactobacillus plantarum ST-III

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Embodiment 1

[0057] Quantitative detection method of plantarum lactobacillus in embodiment 1 probiotic milk products

[0058] a. Preparation of sample total RNA

[0059] Extraction by Trizol method: The fermented milk drink containing Lactobacillus plantarum ST-III (Bright Changyou plant lactic acid bacteria drink) purchased on the market was used as the implementation sample, which were three samples that were 19 days, 15 days and 13 days away from the production date. The sample was ground in a mortar filled with liquid nitrogen, then quickly added 1mL Trizol reagent, according to the instructions of the Trizol kit ( Plus RNA Purification System, U.S. Invitrogen Company, Cat. No. 12183-555) extract sample RNA, then treat twice with DNaseI to ensure that the DNA contamination in the RNA is completely eliminated, obtain a purified RNA sample, measure the concentration, and pass through 1% agar for RNA integrity Sugar gel electrophoresis detection.

[0060] b. Reverse transcription ampli...

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Abstract

The invention discloses a method and a kit for quickly and quantificationally detecting lactobacillus plantarum ST-III. The method comprises the following steps of: 1) extracting total RNA (ribonucleic acid) of a sample to be detected; 2) carrying out inverse transcription on the total RNA extracted in the step 1) to obtain cDNA (complementary deoxyribonucleic acid); 3) carrying out fluorescent quantitative PCR (polymerase chain reaction) amplification detection by utilizing a primer pair of specifically amplified lactobacillus plantarum ST-III and taking the cDNA obtained in the step 2) as atemplate; and 4) comparing the circulation thresholds of the sample to be detected and a quantitative external standard product to quantify the actual number of copies in the sample to be detected. The method and the kit can quickly detect the quantity of lactobacillus plantarum ST-III in a detected sample and have fast speed, strong specificity and high sensitivity, and the limit of detection can be up to 2.85*10<3>CFU/mL.

Description

technical field [0001] The invention belongs to the technical field of biological detection, in particular to a method for rapid quantitative detection of plantarum lactobacillus ST-III and a kit thereof. Background technique [0002] Probiotics are "living microorganisms that, when ingested in certain quantities, can have a beneficial effect on the health of the host". With the development of biotechnology and medicine and other disciplines, the beneficial effects of probiotics on human health have been confirmed more and more. Therefore, probiotic products are increasingly sought after by people, and their production and sales are also on the rise year by year. A large number of studies have shown that only the number of viable bacteria in the probiotics in the product reaches 1×10 6 Only one / ML can play its due effect. However, due to the lack of systematic research and formulation of standards for probiotic products in my country, especially the lack of strict restric...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/06G01N21/64
Inventor 陈臣任婧周方方艾连中
Owner BRIGHT DAIRY & FOOD
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