阅读说明：本技术 一种用于快速检测细胞制品中细菌的培养基及其制备方法和应用 (Culture medium for rapidly detecting bacteria in cell product and preparation method and application thereof ) 是由 申长远 孙振娜 于 2021-07-30 设计创作，主要内容包括：本申请公开了一种用于快速检测细胞制品中细菌的培养基及其制备方法和应用,所述培养基的原料按重量份包括：平衡盐水50-54份、葡萄糖10-13份、维生素4-6份、抗生素3-5份、氨基酸15-20份、Ficoll-Paque缓冲液60-80份、细胞消化液4-8份、PH调整液5-9份以及超纯水30-40份。所述制备方法包括如下步骤：混合得到混合物A、加入抗生素、细胞消化液进行混合、PH调整、混合得到混合物B以及加入Ficoll-Paque缓冲液。所述应用包括如下步骤：细胞培养、离心分离以及过滤。本申请的有益之处在于可便于进行其中细菌的集中过滤处理,便于细菌的集中,从而便于进行快速检测,适用于集中检测,便于进行统计以及研究,使用效果较好。(The application discloses a culture medium for rapidly detecting bacteria in cell products, a preparation method and an application thereof, wherein the culture medium comprises the following raw materials in parts by weight: 50-54 parts of balanced saline, 10-13 parts of glucose, 4-6 parts of vitamin, 3-5 parts of antibiotic, 15-20 parts of amino acid, 60-80 parts of Ficoll-Paque buffer solution, 4-8 parts of cell digestive juice, 5-9 parts of pH adjusting solution and 30-40 parts of ultrapure water. The preparation method comprises the following steps: mixing to obtain mixture A, adding antibiotics and cell digestive juice, mixing, adjusting pH, mixing to obtain mixture B, and adding Ficoll-Paque buffer solution. The application comprises the following steps: cell culture, centrifugation and filtration. The beneficial effects of this application lie in can be convenient for carry on wherein the concentrated filtration of bacterium and handle, the concentration of the bacterium of being convenient for to be convenient for carry out short-term test, be applicable to the concentrated detection, be convenient for make statistics of and research, the result of use is better.)

1. A culture medium for rapid detection of bacteria in a cell preparation, comprising:

the culture medium comprises the following raw materials in parts by weight: 50-54 parts of balanced saline, 10-13 parts of glucose, 4-6 parts of vitamin, 3-5 parts of antibiotic, 15-20 parts of amino acid, 60-80 parts of Ficoll-Paque buffer solution, 4-8 parts of cell digestive juice, 5-9 parts of pH adjusting solution and 30-40 parts of ultrapure water.

2. A medium for the rapid detection of bacteria in a cell preparation according to claim 1, wherein:

the balanced salt water is selected from Hanks liquid and Earle liquid or a mixture of the Hanks liquid and the Earle liquid.

3. A medium for the rapid detection of bacteria in a cell preparation according to claim 1, wherein:

the cell digestive juice is selected from one of trypsin solution, EDTA solution and collagenase solution.

4. A medium for the rapid detection of bacteria in a cell preparation according to claim 1, wherein:

the amino acid is prepared by mixing valine, leucine, isoleucine, threonine, lysine, tryptophan, phenylalanine, methionine, histidine, tyrosine, arginine, cystine (L type) and glutamine.

5. A method for preparing a culture medium according to claim 1, wherein: the preparation method comprises the following steps:

(1) adding balanced brine into a container;

(2) adding a dry powder culture medium into ultrapure water at the temperature of between 20 and 30 ℃, further stirring and mixing to obtain a mixture A;

(3) pouring the mixture A in the step (2) into the container in the step (1) and mixing the mixture A with balanced brine;

(4) adding antibiotics and cell digestive juice into the container on the basis of the step (3), and mixing and stirring;

(5) further adding a pH adjusting solution into the container;

(6) adding ultrapure water into a container, diluting to a required volume, stirring by a stirring device, and mixing to obtain a mixture B;

(7) and finally adding Ficoll-Paque buffer solution into the mixture B without mixing to finish the preparation of the culture medium.

6. The method for preparing a culture medium according to claim 5, wherein:

the dry powder culture medium in the step (2) is formed by uniformly mixing glucose, vitamins and amino acid and is dry powder.

7. The method for preparing a culture medium according to claim 5, wherein:

and (3) when the dry powder culture medium is dispersed into ultrapure water in the step (2), washing the packaging bag with water is needed, so that dry powder traces are prevented from remaining on the packaging bag.

8. The method for preparing a culture medium according to claim 5, wherein:

in the step (6), the mixture B is required to be filtered after being prepared; after the mixture B in the step (7) is added with the Ficoll-Paque buffer solution, sealing is needed when the mixture B is stored, and meanwhile, the storage environment is kept sterile, and the storage temperature is 4 ℃.

9. The method for preparing a culture medium according to claim 5, wherein:

in the steps (1) to (7), all vessels need to be strictly disinfected and sterilized when being proportioned.

10. Use of a culture medium according to claim 1, characterized in that: the application comprises the following steps:

(1) pouring the culture medium into a culture dish, a culture test tube or a culture container, and simultaneously raising the temperature of the culture medium to 20-30 ℃;

(2) then pouring the cells to be cultured into a culture dish, a culture test tube or a culture container for culturing;

(3) placing the culture container in an incubator for culture, wherein in the culture process, external gas supply equipment is used for supplying gas inside the incubator, and meanwhile, during gas supply, disinfection treatment of a gas source is required to avoid the entry of mixed bacteria;

(4) while culturing, adding a supplementary culture medium at regular intervals to provide nutrients;

(5) after culturing for a certain time, detecting cells, pouring liquid in a culture dish, a culture test tube or a culture container into centrifugal equipment, and further performing centrifugal treatment under the centrifugal action of the centrifugal equipment to layer the liquid after culturing the cells so that the cells are precipitated under a Ficoll-Paque buffer solution, and bacteria are suspended in the Ficoll-Paque buffer solution;

(6) the centrifuged Ficoll-Paque buffer solution is filtered by a filter membrane, so that bacteria are filtered, and therefore, the bacteria can be concentrated and can be rapidly detected.

Technical Field

The application relates to the technical field of cell culture, in particular to a culture medium for rapidly detecting bacteria in a cell product, and a preparation method and application thereof.

Background

The culture medium is a nutrient medium prepared from different nutrient substances for the growth and reproduction of microorganisms, plants or animals (or tissues). Generally comprises several major substances such as carbohydrate, nitrogen-containing substances, inorganic salts (including trace elements), vitamins and water. The culture medium is not only a basic substance for providing nutrition for the cells and promoting the proliferation of the cells, but also a living environment for the growth and the propagation of the cells.

The culture medium in the prior art is not easy to be used after cell culture when the cell culture is carried out. Wherein the isolation of bacteria; thereby being difficult to carry out the rapid detection of bacteria therein, the detection effect may be poor, and the use is difficult. Therefore, a culture medium for rapidly detecting bacteria in a cell preparation, a preparation method and application thereof are provided aiming at the problems.

Disclosure of Invention

The embodiment provides a culture medium for rapidly detecting bacteria in a cell product, and a preparation method and application thereof, which are used for solving the problem that cell separation and centralized detection are not easy to perform in the prior art.

According to one aspect of the application, a culture medium for rapidly detecting bacteria in a cell preparation is provided, wherein the culture medium comprises the following raw materials in parts by weight: 50-54 parts of balanced saline, 10-13 parts of glucose, 4-6 parts of vitamin, 3-5 parts of antibiotic, 15-20 parts of amino acid, 60-80 parts of Ficoll-Paque buffer solution, 4-8 parts of cell digestive juice, 5-9 parts of pH adjusting solution and 30-40 parts of ultrapure water.

Further, the balanced salt water adopts Hanks liquid and Earle liquid or the mixture of the two.

Further, the cell digestive juice is selected from one of trypsin solution, EDTA solution and collagenase solution.

Furthermore, the amino acid is formed by mixing valine, leucine, isoleucine, threonine, lysine, tryptophan, phenylalanine, methionine, histidine, tyrosine, arginine, cystine (L type) and glutamine.

As a second aspect of the present application, there is provided a method for preparing a culture medium for rapid detection of bacteria in a cell preparation, the method comprising the steps of:

(1) adding balanced brine into a container;

(2) adding a dry powder culture medium into ultrapure water at the temperature of between 20 and 30 ℃, further stirring and mixing to obtain a mixture A;

(3) pouring the mixture A in the step (2) into the container in the step (1) to mix the mixture A with balanced brine;

(4) adding antibiotics and cell digestive juice into the container on the basis of the step (3), and mixing and stirring;

(5) further adding a pH adjusting solution into the container;

(6) adding ultrapure water into a container, diluting to a required volume, stirring by a stirring device, and mixing to obtain a mixture B;

(7) and finally adding Ficoll-Paque buffer solution into the mixture B without mixing to finish the preparation of the culture medium.

Further, the dry powder culture medium in the step (2) is formed by uniformly mixing glucose, vitamins and amino acid and is in a dry powder shape.

Further, in the step (2), when the dry powder culture medium is dispersed into ultrapure water, the packaging bag needs to be washed with water, so that dry powder traces are prevented from remaining on the packaging bag.

Further, the mixture B in the step (6) needs to be filtered after being prepared; after the mixture B in the step (7) is added with the Ficoll-Paque buffer solution, sealing is needed when the mixture B is stored, and meanwhile, the storage environment is kept sterile, and the storage temperature is 4 ℃.

Further, when the proportioning is performed in the steps (1) to (7), all the vessels need to be strictly sterilized.

As a third aspect of the present application, there is provided a use of a culture medium for rapid detection of bacteria in a cell preparation, the use comprising the steps of:

(1) pouring the culture medium into a culture dish, a culture test tube or a culture container, and simultaneously raising the temperature of the culture medium to 20-30 ℃;

(2) then pouring the cells to be cultured into a culture dish, a culture test tube or a culture container for culturing;

(3) placing the culture container in an incubator for culture, wherein in the culture process, external gas supply equipment is used for supplying gas inside the incubator, and meanwhile, during gas supply, disinfection treatment of a gas source is required to avoid the entry of mixed bacteria;

(4) while culturing, adding a supplementary culture medium at regular intervals to provide nutrients;

(5) after culturing for a certain time, detecting cells, pouring liquid in a culture dish, a culture test tube or a culture container into centrifugal equipment, and further performing centrifugal treatment under the centrifugal action of the centrifugal equipment to layer the liquid after culturing the cells so that the cells are precipitated under a Ficoll-Paque buffer solution, and bacteria are suspended in the Ficoll-Paque buffer solution;

(6) the centrifuged Ficoll-Paque buffer solution is filtered by a filter membrane, so that bacteria are filtered, and therefore, the bacteria can be concentrated and can be rapidly detected.

Through the above-mentioned embodiment of this application, this application's beneficial effect lies in: the application provides a culture medium and preparation method and application thereof can carry out the cultivation of cell product, compare with the conventional art, and it is cultivateing the use back, can be convenient for carry on wherein the concentrated filtration of bacterium and handle, the concentration of the bacterium of being convenient for to be convenient for carry out short-term test, be applicable to concentrated detection, be convenient for make statistics of and research, the result of use is better.

Drawings

In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings needed to be used in the description of the embodiments or the prior art will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without inventive exercise.

FIG. 1 is a flow chart of a method for preparing a culture medium for rapid detection of bacteria in a cell preparation;

FIG. 2 is a flow chart of the application of a culture medium for rapid detection of bacteria in cell preparations.

Detailed Description

In order to make the technical solutions better understood by those skilled in the art, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is obvious that the described embodiments are only partial embodiments of the present application, but not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.

It should be noted that the terms "first," "second," and the like in the description and claims of this application and in the drawings described above are used for distinguishing between similar elements and not necessarily for describing a particular sequential or chronological order. It should be understood that the data so used may be interchanged under appropriate circumstances such that embodiments of the application described herein may be used. Furthermore, the terms "comprises," "comprising," and "having," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.

In this application, the terms "upper", "lower", "left", "right", "front", "rear", "top", "bottom", "inner", "outer", "middle", "vertical", "horizontal", "lateral", "longitudinal", and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings. These terms are used primarily to better describe the present application and its embodiments, and are not used to limit the indicated devices, elements or components to a particular orientation or to be constructed and operated in a particular orientation.

Moreover, some of the above terms may be used to indicate other meanings besides the orientation or positional relationship, for example, the term "on" may also be used to indicate some kind of attachment or connection relationship in some cases. The specific meaning of these terms in this application will be understood by those of ordinary skill in the art as appropriate.

Furthermore, the terms "mounted," "disposed," "provided," "connected," and "sleeved" are to be construed broadly. For example, it may be a fixed connection, a removable connection, or a unitary construction; can be a mechanical connection, or an electrical connection; may be directly connected, or indirectly connected through intervening media, or may be in internal communication between two devices, elements or components. The specific meaning of the above terms in the present application can be understood by those of ordinary skill in the art as appropriate.

It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict. The present application will be described in detail below with reference to the embodiments with reference to the attached drawings.

The culture medium for rapidly detecting bacteria in cell products, the preparation method and the application thereof in the present embodiment can be used in combination with various culture dishes and culture containers, for example, the following culture dishes are provided in the present embodiment, and the culture medium for rapidly detecting bacteria in cell products, the preparation method and the application thereof in the present embodiment can be used in combination with the following culture dishes.

A culture dish, comprising: the dish body is provided with a groove and is used for placing a culture medium; the first cover body comprises a first covering surface and a connecting surface, the connecting surface is arranged at the edge position of the first covering surface and is used for detachably connecting the dish body, and the first covering surface is provided with a slide way which penetrates through the first covering surface; the second cover body is arranged on the upper part of the first cover body and is detachably connected with the first cover body; the inoculation part is detachably connected to the first cover body, and one end of the inoculation part can penetrate through the slide way and is in contact with the culture medium. In the technical scheme provided by the embodiment of the invention, the dish body is provided with a groove for placing a culture medium; the first cover body comprises a first covering surface and a connecting surface, the connecting surface is arranged at the edge position of the first covering surface and is used for detachably connecting the dish body, and the first covering surface is provided with a slide way which penetrates through the first covering surface; the second cover body is arranged at the upper part of the first cover body and is detachably connected with the first cover body; the inoculation part can be dismantled and connect in first lid, and the one end of inoculation part can pass the slide and contact each other with the culture medium, for prior art, need open a seam of culture dish, and the culture medium can contact the bacterium in the air this moment to, when using inoculating the microorganism of inoculating the loop-marking method, need the left hand to open a seam of culture dish, the right hand is drawn the line and is inoculated. The operation beginner is not easy to master, the culture medium is easy to contact with the air in a large area, and therefore pollution is caused, and the flat plate scribing method needs skilled operation, and a beginner is easy to scratch an agar layer by manual scribing to cause damage of the culture medium. The dish body is used for supporting the first cover body and the second cover body, the dish body is provided with a groove for placing a culture medium, the dish body is usually made of transparent glass materials or plastic materials, and the bottom of the groove is a plane, so that the culture medium is convenient to place; the first cover body comprises a first covering surface and a connecting surface, the connecting surface is arranged at the edge position of the first covering surface and is used for detachably connecting the dish body, a slide way is arranged on the first covering surface and penetrates through the first covering surface, the first cover body is usually also made of transparent glass material or plastic material, the culture medium in the dish body can be conveniently observed, the connecting surface can be in mutual contact with the dish body, for example, an internal thread is arranged on the connecting surface, an external thread is arranged on the dish body, so that the first cover body can be connected with the dish body, of course, the dish body and the first cover body can be detachably connected in other modes, for example, clamping connection can be carried out, or the first cover body is buckled on the dish body, therefore, the connecting surface can be connected with the dish body as long as the connecting surface can be connected with the dish body, the angle between the first covering surface and the connecting surface is usually a degree, the first covering surface is horizontally arranged, the connecting surface is vertical to the first covering surface, the slide way penetrates through two ends of the first covering surface, and the slide way is in various shapes, such as a strip shape or an arc shape, on the first covering surface, and the main purpose is to facilitate scribing inoculation; the second cover body is arranged at the upper part of the first cover body and is detachably connected with the first cover body; the inoculation part is detachably connected to the first cover body, one end of the inoculation part can penetrate through the slide way and is in contact with the culture medium, and the second cover body is used for covering the slide way so as to prevent dust from entering the dish body through the slide way; in the embodiment of the invention, the slide way is arranged on the first cover body and penetrates through the first cover body, one end of the inoculation part can penetrate through the slide way and is in contact with the culture medium, so that the culture medium can be conveniently streaked and inoculated by personnel through the inoculation part, only the slide way of the first cover body is exposed in the air, the contact area of the culture medium and the air is reduced, and the second cover body is detachably connected with the driver cover body, so that the slide way of the second cover body can be covered through the second cover body, and the technical effect of protecting the culture medium is achieved. Furthermore, the slide way comprises a through hole and a slide rail, the through hole penetrates through the first covering surface, and the slide rail is arranged at the edge of the through hole. In this embodiment, a slide way is further defined, the through hole penetrates through two sides of the first covering surface, so that one end of the inoculating part can penetrate through the through hole and contact with the culture medium, meanwhile, the inoculating part can move along the direction of the slide rail, so that the depth of the inoculating part inserted into the culture medium is fixed after one end of the inoculating part contacts with the culture medium, and therefore streaking inoculation is convenient for personnel. Further, the inoculation part comprises a penholder and a rotating body, wherein the penholder is provided with threads, and the rotating body is rotatably connected with the penholder. In this embodiment, further limited the inoculation part, the pen-holder adopts plastics or glass material usually, the rotator adopts plastics material usually, the outside of pen-holder sets up the external screw thread, the middle part of rotator has the through-hole, the edge of through-hole sets up the internal thread, make the rotator rotate and connect in the pen-holder, after the perforation was passed to the one end of pen-holder, adjust the distance between pen-holder and the culture medium through rotating the rotator, then place the bottom of rotator on the slide rail, make the one end and the culture medium of pen-holder contact each other, and, the degree of depth that the pen-holder inserted the culture medium is unchangeable all the time, thereby reach the technological effect of making things convenient for the streak inoculation. Further, the inoculation part also comprises an inoculating loop, and the inoculating loop is detachably connected to the penholder. In the embodiment, an inoculating part is further defined, the inoculating loop is an inoculating tool commonly used in bacterial culture and widely applied to various fields of subjects such as microbial detection, cell microbiology, molecular biology and the like, in the scribing method, the inoculating loop is generally used for adhering bacteria-containing materials and scribing on the surface of a solid culture medium, the inoculating loop is generally made of plastic or metal materials, and the inoculating loop is detachably connected with a penholder, such as in plug-in connection or threaded connection, so that the installing and the disassembling of the inoculating loop are facilitated. Further, increase joint part, joint part includes buckle and draw-in groove, and the buckle setting is at the border position of second lid, and the draw-in groove setting is at the border position of first lid, and the buckle can insert in the draw-in groove. In this embodiment, a clamping component is added, and the clamping component is used for connecting the first cover body and the second cover body, a buckle is arranged at the edge of the second cover body, and a clamping groove is arranged at the edge of the first cover body, so that the buckle can be clamped into the clamping groove, thereby achieving the technical effect of connecting the first cover body and the second cover body; at this moment, the shape of the connectable part is 'L' shape or semi-circular arc shape, and certainly, a protrusion can be arranged at a position where the buckle is close to the clamping groove, so that the protrusion is clamped into the clamping groove, thereby achieving the technical effect of conveniently connecting and fixing the first cover body and the second cover body. Furthermore, the clamping grooves comprise a first clamping groove and a second clamping groove, the first clamping groove is arranged at the edge position of the first cover body, the second clamping groove is arranged at the edge position of the first cover body, and two ends of the buckle can be respectively inserted into the first clamping groove and the second clamping groove. In this embodiment, further limited the draw-in groove, the draw-in groove is two, is first draw-in groove and second draw-in groove respectively, and first draw-in groove sets up the border position at first lid, and the second draw-in groove sets up the border position at first lid, and at this moment, the buckle is not connected with any part, but sets up alone, then blocks first draw-in groove and second draw-in groove with the both ends card simultaneously of buckle to reach the effect of fixed first lid and second lid, at this moment, the shape of buckle is "U" shape structure. Further, the second cover body is detachably connected to the dish body. In the embodiment, the second cover body is further limited, and the second cover body is detachably connected to the dish body, so that the technical effect of sealing the groove is achieved; optionally, an internal thread is arranged on the second cover body, an external thread is arranged on the side wall of the dish body, so that the second cover body is in threaded connection with the dish body, and at the moment, the first cover body is buckled on the dish body and is positioned between the second cover body and the dish body.

Of course, this embodiment can be applied to a culture dish or culture container having other structures. Not to be repeated, a culture medium for rapidly detecting bacteria in a cell preparation, a preparation method thereof, and applications thereof according to embodiments of the present application are described below.

Referring to fig. 1-2, a culture medium for rapid detection of bacteria in cell preparations is provided, the culture medium comprises the following raw materials in parts by weight: 50-54 parts of balanced saline, 10-13 parts of glucose, 4-6 parts of vitamin, 3-5 parts of antibiotic, 15-20 parts of amino acid, 60-80 parts of Ficoll-Paque buffer solution, 4-8 parts of cell digestive juice, 5-9 parts of pH adjusting solution and 30-40 parts of ultrapure water.

Further, the balanced salt water adopts Hanks liquid and Earle liquid or the mixture of the two.

Further, the cell digestive juice is selected from one of trypsin solution, EDTA solution and collagenase solution.

Furthermore, the amino acid is formed by mixing valine, leucine, isoleucine, threonine, lysine, tryptophan, phenylalanine, methionine, histidine, tyrosine, arginine, cystine (L type) and glutamine.

Further provided is a method for preparing a culture medium for rapid detection of bacteria in a cell preparation, the method comprising the steps of:

(1) adding balanced brine into a container;

(2) adding a dry powder culture medium into ultrapure water at the temperature of between 20 and 30 ℃, further stirring and mixing to obtain a mixture A;

(3) pouring the mixture A in the step (2) into the container in the step (1) to mix the mixture A with balanced brine;

(4) adding antibiotics and cell digestive juice into the container on the basis of the step (3), and mixing and stirring;

(5) further adding a pH adjusting solution into the container;

(6) adding ultrapure water into a container, diluting to a required volume, stirring by a stirring device, and mixing to obtain a mixture B;

(7) and finally adding Ficoll-Paque buffer solution into the mixture B without mixing to finish the preparation of the culture medium.

Further, the dry powder culture medium in the step (2) is formed by uniformly mixing glucose, vitamins and amino acid and is in a dry powder shape.

Further, in the step (2), when the dry powder culture medium is dispersed into ultrapure water, the packaging bag needs to be washed with water, so that dry powder traces are prevented from remaining on the packaging bag.

Further, the mixture B in the step (6) needs to be filtered after being prepared; after the mixture B in the step (7) is added with the Ficoll-Paque buffer solution, sealing is needed when the mixture B is stored, and meanwhile, the storage environment is kept sterile, and the storage temperature is 4 ℃.

Further, when the proportioning is performed in the steps (1) to (7), all the vessels need to be strictly sterilized.

Still further provided is a use of a culture medium for rapid detection of bacteria in a cell preparation, the use comprising the steps of:

(1) pouring the culture medium into a culture dish, a culture test tube or a culture container, and simultaneously raising the temperature of the culture medium to 20-30 ℃;

(2) then pouring the cells to be cultured into a culture dish, a culture test tube or a culture container for culturing;

(3) placing the culture container in an incubator for culture, wherein in the culture process, external gas supply equipment is used for supplying gas inside the incubator, and meanwhile, during gas supply, disinfection treatment of a gas source is required to avoid the entry of mixed bacteria;

(4) while culturing, adding a supplementary culture medium at regular intervals to provide nutrients;

(5) after culturing for a certain time, detecting cells, pouring liquid in a culture dish, a culture test tube or a culture container into centrifugal equipment, and further performing centrifugal treatment under the centrifugal action of the centrifugal equipment to layer the liquid after culturing the cells so that the cells are precipitated under a Ficoll-Paque buffer solution, and bacteria are suspended in the Ficoll-Paque buffer solution;

(6) the centrifuged Ficoll-Paque buffer solution is filtered by a filter membrane, so that bacteria are filtered, and therefore, the bacteria can be concentrated and can be rapidly detected.

The application has the advantages that:

the application provides a culture medium and preparation method and application thereof can carry out the cultivation of cell product, compare with the conventional art, and it is cultivateing the use back, can be convenient for carry on wherein the concentrated filtration of bacterium and handle, the concentration of the bacterium of being convenient for to be convenient for carry out short-term test, be applicable to concentrated detection, be convenient for make statistics of and research, the result of use is better.

The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.