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Detection kit and detection method for beta-lactamase in raw milk and milk products

A detection kit and a technology for lactamase, which are applied in the directions of microorganism-based methods, microorganism determination/inspection, biochemical equipment and methods, etc., can solve the problems of complicated experimental operation, unfavorable grass-roots laboratory adoption, etc., and achieve the sensitivity of results. High, convenient and fast operation, low cost effect

Inactive Publication Date: 2009-09-30
河南省兽药监察所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the cup-and-saucer method recommended by it has disadvantages such as relatively complicated experimental operation, a series of drug standard solutions, the need to prepare culture medium and make it into a plate for inspection, and the concentration of bacteria liquid needs to be tested in advance, which is not conducive to the adoption of basic-level laboratories.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] 1) 5 parts of blank milk were added with β-lactamase to prepare samples with the following contents: 3U / mL, 5U / mL, 10U / mL, 15U / mL, 30U / mL. And set pure water as a blank control.

[0058] 2) β-lactamase standard dilution is 3000U / mL.

[0059] 3) Take out the vacuum-packed sterilized culture dish that can be stored for a long time. The vacuum-packed sterilized culture dish that can be stored for a long time has been coated with Bacillus stearothermophilus, and paste 13mm blank test paper sheets in the order of A, B, C, and D. On the surface of a sterilized petri dish that can be stored in a vacuum package for a long time.

[0060] 4) Take two freeze-dried vials containing 50 ng of ampicillin and 50 ng of ampicillin + 10 ug of sulbactam, respectively, and add 1 mL of milk containing β-lactamase in different doses. The lyophilized vials of the drug are marked as A and C respectively, and the lyophilized vials of the drug containing ampicillin + sulbactam are marked as B a...

Embodiment 2

[0067] 1) Dilute the β-lactamase standard to 3000U / mL.

[0068] 2) Take 5 parts of blank milk, add β-lactamase, and prepare samples with the following contents: 3U / mL, 10U / mL, 30U / mL, 100U / mL, 300U / mL. And set pure water as a blank control.

[0069] 3) Take the vacuum-packed and long-term-preserved sterilized petri dish, which has been coated with Bacillus stearothermophilus, and paste the blank test paper on the Petri dish surface.

[0070] 4) Take two bottles of freeze-dried vials containing 50 ng of ampicillin and 50 ng of ampicillin + 20 μg of sulbactam, and add 1 mL of β-lactamase milk with different doses, and the ampicillin-containing ampicillin The bottles are marked as A and C respectively, and the vials containing ampicillin + sulbactam are marked as B and D. Add 10ul of 3000U / mL β-lactamase to bottles C and D respectively. And blankly take 50 μL of the contents of the freeze-dried vials marked A, B, C, D and pure water samples, and add them to the blank test pape...

Embodiment 3

[0076] 1) Dilute the β-lactamase standard to 3000U / mL.

[0077] 2) Take 5 parts of blank yogurt, adjust its pH to 7.0, add diluted β-lactamase, and prepare samples with the following content: 3U / mL, 30U / mL, 100U / mL, 300U / mL, 3000U / mL . And set pure water as a blank control.

[0078] 3) Take the vacuum-packed and long-term-preserved sterilized petri dish, which has been coated with Bacillus stearothermophilus, and paste the blank test paper on the Vacuum packaging allows for long-term preservation of sterilized Petri dish surfaces.

[0079] 4) Take two bottles of freeze-dried vials containing 50 ng of ampicillin and 50 ng of ampicillin + 20 μg of sulbactam, respectively, and add 1 mL of β-lactamase yogurt to which different doses have been added. The dry vials are marked as A and C respectively, and the freeze-dried vials containing ampicillin + sulbactam are marked as B and D. Add 10ul of 3000U / mL β-lactamase to the vials C and D respectively. Place in a 36°C±1°C incubator a...

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Abstract

The invention relates to a detection kit and a detection method of raw milk and milk products, in particular to a detection kit and a detection method for beta-lactamase in raw milk and milk products. At present, no diary corporation in China can carry out production with the raw milk completely supplied by self-owned farms. All diary corporations need to purchase raw milk from dairy farmer to some extent. Thus, the detection of imparities and antibiotics becomes extremely important. The detection method adopts a standard strain of bacillus stearothermophilus absolutely sensitive to penicillin antibiotics, uses the specificity of sulbactam to inhibit the activity of the beta-lactamase, adds ampicillin as a reference and contrasts inhabitation zones of a sample with a beta-lactamase inhibitor and a sample without the inhibitor to detect the antibiotics. The detection kit comprises a sterilized culture dish with a vacuum package, a beta-lactamase standard product, a freeze-drying cillin bottle containing ampicillin and ampicillin and sulbactam, a blank filter paper sheet and a diluent. The detection kit and the detection method have the advantages of low cost, convenient and quick operation and high sensitivity.

Description

technical field [0001] The invention relates to a detection kit and detection method for fresh milk and dairy products, in particular to a detection kit and detection method for β-lactamase in raw fresh milk and dairy products. Background technique [0002] Milk is rich in nutrition, easy to digest and absorb, high in quality and low in price, and convenient to eat. It is the most "close to perfect food". It is called "white blood" and is the most ideal natural food. [0003] The protein in milk is mainly casein, albumin, globulin, milk protein, etc. Among the more than 20 kinds of amino acids contained in it, there are 8 kinds of amino acids necessary for the human body. Milk protein is a full-priced protein, and its digestibility is as high as 98%. . Milk fat is a high-quality fat with the best quality. Its digestibility is above 95%, and it contains a lot of fat-soluble vitamins. Lactose in milk is galactose and lactose, which are the most easily digested and absorbed s...

Claims

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Application Information

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IPC IPC(8): C12Q1/34C12Q1/18C12R1/07
Inventor 班付国刘宏伟高延玲黄克炎张发旺周红霞崔清兰马俊
Owner 河南省兽药监察所
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