阅读说明：本技术 一种微生物凝胶琼脂培养基 (Microbial gel agar culture medium ) 是由 孙百莉 于 2020-07-22 设计创作，主要内容包括：本发明公开了一种微生物凝胶琼脂培养基。利用本发明制作的凝胶琼脂平板可吸收1mL样液,能直接用于样品检测,操作简便,检测效率高；整个接种过程均在常温条件下进行,不耐热菌和受损伤菌不会因为热激而死亡；有抑菌作用样品,高渗透压、高酸碱性样品可在培养基中扩散,有效稀释样品中的抑菌因素,检测结果与常规方法接近；接种后样品中的稀释液渗透到培养基中,样品中的微生物则留在平板表面,氧气充足,利于需氧和兼性厌氧菌的生长。(The invention discloses a microbial gel agar culture medium. The gel agar plate prepared by the invention can absorb 1mL of sample liquid, can be directly used for sample detection, and has simple and convenient operation and high detection efficiency; the whole inoculation process is carried out at normal temperature, and thermolabile bacteria and damaged bacteria can not die due to heat shock; the sample with the bacteriostatic action, the sample with high osmotic pressure and high acidity and alkalinity can be diffused in the culture medium, the bacteriostatic factors in the sample can be effectively diluted, and the detection result is close to that of the conventional method; after inoculation, the diluent in the sample permeates into the culture medium, and microorganisms in the sample are remained on the surface of the plate, so that sufficient oxygen is provided, and the growth of aerobic and facultative anaerobes is facilitated.)

1. A microbial gel agar culture medium is characterized in that the culture medium comprises a conventional agar culture medium and gel.

2. The microbial gel agar medium of claim 1, wherein the gel is one of xanthan gum, sodium alginate, guar gum, or locust bean gum.

3. The gel of claim 1 in an amount of 10g to 50 g/L.

4. The regular agar medium of claim 1 is a PCA medium, a VRBA-MUG medium, an XLD medium, a MAC medium, a HE medium, a PDA medium, a PALCAM medium, a CT-sMAC medium, a TCBS medium, a MYP medium, or a VRBGA medium.

5. A method of microbial enumeration comprising the steps of:

1) preparing a solution of the microbial gel agar medium of claims 1-4 by adding water, autoclaving, cooling to about 50 ℃, and plating to obtain a gel agar plate;

2) dripping a microbial sample solution to be detected on the surface of the gel agar plate, uniformly mixing along a tabletop, standing for 30min to ensure that the sample solution is completely absorbed by a culture medium, and obtaining an inoculated gel agar plate;

3) and placing the inoculated gel agar plate into an incubator for culture, and counting the cultured microorganisms.

Technical Field

The invention relates to a culture medium, in particular to a microbial gel agar culture medium.

Background

At present, a flat plate counting method is mainly adopted for detecting the microbial limit, and because 1mL of sample liquid cannot be completely absorbed after agar is solidified, a uniform mixing method is adopted when the limit of the hygiene index bacteria in a sample is detected, namely 1mL of sample liquid is absorbed and added into a culture dish, and then agar culture medium which is sterilized and cooled to about 45 ℃ is poured. The agar culture medium can only be prepared at present and cannot be produced commercially; a detector needs to do a large amount of preparation work, including brushing, culture medium preparation, sterilization, waste disposal and the like, so that the detection efficiency is low; the operator is required to strictly control the temperature, when the temperature is too high, thermolabile microorganisms and damaged microorganisms are easy to die, the detection value is low, and when the temperature is too low, agar is easy to solidify. The limit detection of pathogenic bacteria adopts a plate coating method, namely 1mL of sample liquid is coated on 3 separate plates, the detection efficiency is low, and the cost is high.

The quick detection product is favored by detectors because the preparation time of operators is saved and the detection efficiency is improved. The quick detection products for detecting the microbial limit in the current market are mainly quick detection paper sheets and gel test sheets, the two products directly absorb 1mL of sample liquid to be dripped on the paper sheets and the test sheets, and the sample liquid is not diluted, so that samples with sterilization and bacteriostasis effects and high osmotic pressure and high-acidity and alkalinity samples are detected, and the detection value is lower.

Disclosure of Invention

In order to solve the problems, the invention provides a microbial gel agar culture medium, the culture medium contains gel, a gel agar plate prepared by using the gel agar culture medium can absorb 1mL of sample liquid, can be directly used for sample detection, and has the advantages of convenient use and accurate detection result.

In order to achieve the technical purpose and achieve the technical effect, the invention is realized by the following technical scheme:

the microorganism gel agar culture medium is prepared by adding 10-50 g cold water soluble gel per liter on the basis of agar culture medium, autoclaving, cooling to about 50 deg.C, and pouring into flat plate.

The cold water soluble gel is one of xanthan gum, sodium alginate, guar gum or locust bean gum.

The agar culture medium is PCA culture medium, VRBA-MUG culture medium, XLD culture medium, MAC culture medium, HE culture medium, PDA culture medium, PALCAM culture medium, CT-sMAC culture medium, TCBS culture medium, MYP culture medium or VRBGA culture medium.

In the examples of the present invention, the gel agar medium used was a PCA gel agar medium and a Baird-Parker gel agar medium.

The application of the microbial gel agar culture medium in microbial detection is also within the protection scope of the invention.

The invention provides a method for detecting microorganisms.

The method provided by the invention comprises the following steps:

1) preparation of gel agar plates

Preparing a gel agar culture medium into a solution, cooling to about 50 ℃ after autoclaving, pouring into a flat plate, and pouring 15mL-20 mL/dish;

2) inoculation of

Inoculating the prepared sample solution by selecting 2-3 appropriate dilutions, sucking 1mL of sample solution, dripping the sample solution on the surface of a gel agar plate, uniformly mixing along a tabletop, standing for 30min to ensure that the sample solution is completely absorbed by a culture medium, and repeating 2 dilutions;

3) culturing

Culturing the inoculated flat plate according to the temperature and time required by the detection standard;

4) counting

And taking out the plate after the culture is finished, and counting according to the detection standard requirement.

Compared with the prior art, the invention has the beneficial effects that:

the gel agar plate prepared by the invention can absorb 1mL of sample solution and can be directly used for sample detection; the batch filling can be carried out by a filling machine; a user directly drops 1mL of sample solution on the surface of the gel agar plate, the operation is simple and convenient, and the detection efficiency is high; the whole inoculation process is carried out at normal temperature, and thermolabile bacteria and damaged bacteria can not die due to heat shock; in the limit detection of pathogenic bacteria microorganisms, 1mL of sample liquid can be absorbed by one flat plate, so that the cost is reduced; the content of the culture medium in the flat plate is 15-20 mL, the sample has an antibacterial effect, the sample with high osmotic pressure and high acidity and alkalinity can be diffused in the culture medium, the antibacterial factors in the sample can be effectively diluted, and the detection result is close to that of the conventional method; after inoculation, the diluent in the sample permeates into the culture medium, and microorganisms in the sample are remained on the surface of the plate, so that sufficient oxygen is provided, and the growth of aerobic and facultative anaerobes is facilitated.

Detailed Description

The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.

Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.