Escherichia coli detection kit as well as preparation method and application thereof

A detection kit and Escherichia coli technology, applied in biochemical equipment and methods, methods based on microorganisms, measurement/testing of microorganisms, etc., can solve problems such as inconvenient operation, long detection time, and large limitations in the field of use. To achieve the effect of improving detection efficiency

Pending Publication Date: 2019-03-29
长春万成生物电子工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These two products need to be sampled during use. Using a constant temperature incubator to cultivate for 24 hours is not only inconvenient to operate, but also takes a long time to detect (more than 24 hours). Both products are prepared using the principle of microbial detection (national standard) method
Moreover, the use of the above-mentioned products is limited and can only be used for microbiological testing on the surface of food and tableware, or testing of pathogenic Escherichia coli, which does not have universal applicability.

Method used

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  • Escherichia coli detection kit as well as preparation method and application thereof
  • Escherichia coli detection kit as well as preparation method and application thereof
  • Escherichia coli detection kit as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: kit of the present invention

[0032] 1. Lysis solution

[0033] Add water according to the concentration in any row in Table 1 to prepare the lysate.

[0034] Table 1

[0035]

[0036]

[0037] 2. Test strips

[0038] Weigh 0.02g of X-GAL, add 0.02g of polyvinylpyrrolidone, and add 100mL of absolute ethanol to obtain a color developing solution.

[0039] Prepare neutral chromatographic filter paper or any other kind of test paper, soak the test paper in the above-prepared color solution, and dry it at a temperature between normal temperature and 100°C to obtain a color block; cut the color block into (2-6mm )×(2-8mm) is fixed to one end of the substrate through double-sided adhesive tape to obtain a test strip. The structure diagram is shown in figure 1 .

[0040] 3. Pack test strips, lysate, reaction tubes, straws, and desiccant into a kit.

Embodiment 2

[0041] Embodiment 2: detection test of kit of the present invention

[0042] Select Escherichia coli bacteria solution with OD value of 0.3, sterile water, mold solution with OD value of 0.3 and staphylococcus bacteria solution with OD value of 0.3 as test objects for detection;

[0043]Add the lysate 4 in Example 1 to the water sample to be tested, and let it stand for 10-20min; then use the detection test strip in the kit to immerse the water sample to be tested, keep it for 10min, if the detection test strip shows Any degree of blue is positive, and the test strip is colorless or other colors are negative.

[0044] The test results are shown in Table 2 and figure 2 ;

[0045] Table 2

[0046] Sample name

[0047] It can be seen from the above results that only the E. coli bacterial liquid changes color and shows blue, and the other three groups have no color development. At the same time, the whole process only takes 20 minutes (10min cracking+10min reaction),...

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Abstract

The invention relates to the technical field of water quality microbiological detection and discloses an escherichia coli detection kit as well as a preparation method and an application thereof. Thekit comprises a lysate and a detection test strip, wherein the lysate is prepared from a dehydrant, a penetrant and a surfactant, and the dehydrant is sodium chloride, EDTA-2Na or sodium hydroxide; the penetrant is glycerin, polyethylene glycol 2000, Tween80 or triton-100; the surfactant is polyvinylpyrrolidone or ethyl cellulose; a color developing block is arranged at one end of the detection test strip and contains X-GAL. With the adoption of a dry chemical test paper method, escherichia coli cracks to release galactase and is subjected to a reaction with X-GAL (a galactoside color developing agent) to show blue color, so that dry chemical test paper is prepared; the overall detection process can be shortened to be within 30 min through combination with the specific rapid lysate, and compared with at least 24 h detection time of an existing product, detection efficiency of the escherichia coli in water and food is remarkably improved.

Description

technical field [0001] The invention relates to the technical field of water quality microorganism detection, in particular to an Escherichia coli detection kit and a preparation method and application thereof. Background technique [0002] Escherichia coli is usually called Escherichia coli, which was discovered by Escherich in 1885. Escherichia coli is a Gram-negative bacterium with a size of 0.5×1-3 microns, perinatal flagella, can move, and has no spores. It can ferment a variety of sugars to produce acid and gas. It is a normal resident bacterium in the intestinal tract of humans and animals. Babies enter the intestinal tract with breastfeeding after birth and accompany people for life, accounting for almost 1 / 3 of the dry weight of feces. [0003] The state stipulates that the total number of colonies per milliliter of drinking water is less than 100, and the total coliform group must not be detected in every 100 milliliters of water. [0004] The number of coliform b...

Claims

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Application Information

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IPC IPC(8): C12Q1/34C12Q1/10C12R1/19
CPCC12Q1/10C12Q1/34
Inventor 赵钢唐春王竑婷
Owner 长春万成生物电子工程有限公司
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