CN111321068A - Detection device for disinfection effect of disinfectant and rapid evaluation method - Google Patents
Detection device for disinfection effect of disinfectant and rapid evaluation method Download PDFInfo
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- CN111321068A CN111321068A CN202010261363.6A CN202010261363A CN111321068A CN 111321068 A CN111321068 A CN 111321068A CN 202010261363 A CN202010261363 A CN 202010261363A CN 111321068 A CN111321068 A CN 111321068A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/34—Internal compartments or partitions
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/38—Caps; Covers; Plugs; Pouring means
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/02—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by impregnation, e.g. using swabs or loops
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
- C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/18—Testing for antimicrobial activity of a material
- C12Q1/20—Testing for antimicrobial activity of a material using multifield media
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/21—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Pseudomonadaceae (F)
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/24—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
- G01N2333/245—Escherichia (G)
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/305—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Micrococcaceae (F)
- G01N2333/31—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Micrococcaceae (F) from Staphylococcus (G)
Abstract
The invention discloses a detection device for the disinfection effect of a disinfectant and a rapid evaluation method, wherein the detection device comprises a base box and a top cover matched with the base box, a plurality of culture lattices for placing a culture medium are arranged in the base box, and a plurality of dilution lattices are arranged on the top cover; an inoculator is also included for inoculating the diluted grid sample into the culture medium of the grid. The detection device can be used for rapidly, qualitatively and quantitatively detecting microorganisms and conveniently and rapidly evaluating the disinfection effect of the disinfectant.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a detection device for the disinfection effect of a disinfectant and a rapid evaluation method.
Background
The biosafety of the large-scale farm is the basis for guaranteeing the virtuous cycle of poultry industry, improving the breeding benefit and realizing high-quality, high-efficiency and sustainable development. The disinfection is an important guarantee for eliminating infection sources and cutting off epidemic disease transmission ways, is one of means for effectively preventing and controlling the epidemic diseases, and the disinfectant with good characteristics can effectively control the environmental sanitation of the poultry house, improve the health of animals, increase the income of the breeding industry and the like. Therefore, the evaluation of the disinfection effect of the disinfectant also becomes an important link in the disinfection work.
The current domestic rapid disinfection effect evaluation technical system is still imperfect and comprises a traditional colony counting evaluation method and a modern biological technology. The traditional method comprises an air sedimentation bacteria method and a suspension quantitative sterilization test, and the technology is a classical method for detecting living microorganisms, and the detection result is accurate. However, the traditional method needs to prepare different selective culture media at the same time, so that the experiment consumes more material, wastes time and labor, has slow result production and higher requirement on experiment operation, and influences the development of subsequent work.
Modern biological techniques include immunological methods, fluorescence techniques and PCR methods. The immunological method uses target microbe or its surface structure as direct or indirect antigen, and utilizes the reaction of specific antibody to detect microbe qualitatively or quantitatively. However, this operation is complicated, and the specificity of the antigen-antibody reaction is not high when it is directed to a specific microorganism, and the sensitivity is not high. After some microorganisms are killed by the disinfection factor, the antigenicity of the microorganisms still exists, and a false positive result appears in the effect evaluation; in the fluorescence technique, a target cell is first fluorescently labeled with a nucleic acid probe, a specific enzyme, a specific antibody, or the like, and then quantitatively analyzed by laser scanning. The cell activity is conveniently measured by using the fluorescent probe, but the fluorescent signal is not stable enough and is greatly influenced by background fluorescence, and the sensitivity is also limited by the content of target microorganisms. In addition, the cost is high, the operation is complex, the requirement on the experimental operation environment is high, and the method is not easy to be used in a farm daily; the PCR technique can rapidly detect and detect microorganisms with high sensitivity, but since the half-life of DNA in dead bacterial cells is widely varied, the PCR method cannot distinguish between live cells and dead cells, and there are many problems in evaluating the sterilization effect using the same. In addition, the PCR technology also needs primer design, sample DNA extraction, specific sequence amplification, electrophoresis analysis or gene probe detection, and has complex operation process and high cost.
Disclosure of Invention
The invention aims to provide a detection device and a rapid evaluation method for the disinfection effect of a disinfectant, and aims to solve the problems of high operation technical requirements, complex operation process, high cost, unstable detection results, false positives, time and labor waste and large material consumption of modern biological methods.
The technical scheme adopted by the invention is as follows: a detection device for the disinfection effect of a disinfectant comprises a base box and a top cover matched with the base box, wherein a plurality of culture lattices for placing a culture medium are arranged in the base box, and a plurality of dilution lattices are arranged on the top cover; further comprising an inoculator for inoculating the sample of the diluted grid into the culture medium of the incubation grid.
Preferably, the inoculator comprises an inoculating plate, and a plurality of groups of inoculating needles are arranged on the inoculating plate.
Preferably, the base box is provided with M rows and N columns of culture grids, the top cover is provided with M rows and N columns of dilution grids, the culture grids and the dilution grids are the same in size, the inoculation plate is provided with M or N groups of inoculation needles arranged side by side, and M and N are positive integers.
Preferably, the culture medium components of the culture lattices in each row in the medium box are different.
Preferably, the medium includes at least one of a CN medium, a BP medium, and an EMB medium.
Preferably, each group of inoculating needles comprises X rows and Y rows of inoculating needles, each inoculating needle is in a conical shape, and X and Y are positive integers.
Preferably, each inoculating needle is 8mm in length and 0.6mm in diameter at the end remote from the tip.
The other technical scheme adopted by the invention is as follows: a rapid evaluation method for the disinfection effect of a disinfectant comprises the detection device, and comprises the following steps:
collecting samples: collecting samples before and after disinfection by a disinfectant;
diluting: PBS buffer solution is filled in a dilution grid in the top cover, and each sample is diluted by the PBS buffer solution according to concentration gradient;
inoculation: inoculating each diluted sample into the medium in the cartridge with an inoculator;
culturing: culturing in 37 deg.C incubator for 12-24 hr;
and (5) counting results: and counting the number of bacterial colonies of each culture medium, and evaluating the disinfection effect of the disinfectant according to the comparison result of the number of bacterial colonies before and after disinfection of the disinfectant.
Preferably, the sample collection comprises the following processes: if the sample is solid, the sample is firstly sucked and washed by PBS buffer solution, then washing liquid is collected, sediment is collected after centrifugation, and the sediment is dispersed in the PBS buffer solution again to obtain the sample before and after disinfection by the disinfectant; if the sample is liquid, carrying out concentration treatment according to the turbidity degree of the sample to obtain the sample before and after disinfection by the disinfectant; if the sample is air, the detection device can be directly used for collecting the sample by adopting a natural sedimentation method, an air sampler can be used for pumping and filtering the air, bacteria and dust in the air can be collected into PBS buffer solution after impacting the water surface, and then the sample is concentrated to obtain the sample before and after being disinfected by the disinfectant.
Preferably, the inoculation comprises the following steps: each diluted sample was dipped into the medium in the media cassette using an inoculator and triplicate samples were taken for each diluted sample.
The invention has the beneficial effects that:
the invention designs a novel detection device with disinfectant disinfection effect by improving the traditional detection device, and can simultaneously and qualitatively detect pseudomonas aeruginosa, escherichia coli and staphylococcus aureus. The components in the culture medium in the culture lattices of the detection device can effectively inhibit the growth of the mould and have higher selectivity on the three detected target strains. The dilution grid is used for detecting samples with different dilutions and quantitatively detecting microorganisms. The design of upper cover is used for diluting the sample in the detection device and uses, practices thrift the cost, need not independent dilution sample container, makes things convenient for the field usage in plant. The design of the inoculating needle on the inoculator can quickly inoculate the sample and save the packaging space. The detection device can be used for rapidly, qualitatively and quantitatively detecting microorganisms and conveniently and rapidly evaluating the disinfection effect of the disinfectant.
Drawings
FIG. 1 is a schematic structural diagram of a device for detecting the disinfection effect of a disinfectant according to the present invention;
FIG. 2 is a schematic structural diagram of a base box of the device for detecting the disinfection effect of a disinfectant according to the present invention;
FIG. 3 is a schematic view showing the structure of an inoculator in the device for detecting the disinfection effect of a disinfectant according to the present invention;
fig. 4 is an operation flowchart of a method for rapidly evaluating the sterilization effect of a disinfectant according to the present invention;
FIG. 5 is a graph showing the result of a rapid evaluation method for the sterilization effect of a disinfectant according to the present invention;
in the figure: 1-a base box; 2-a top cover; 3-cultivating lattices; 4-dilution of grids; 5-inoculating a plate; 6-inoculating needle.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention. The components of the embodiments of the present invention generally described and illustrated in the figures herein may be arranged and designed in a wide variety of different configurations.
Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. 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 invention.
It should be noted that the embodiments and features of the embodiments may be combined with each other without conflict.
It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, it need not be further defined and explained in subsequent figures.
In the description of the embodiments of the present invention, it should be noted that the indication of the orientation or the positional relationship is based on the orientation or the positional relationship shown in the drawings, or the orientation or the positional relationship which is usually placed when the product of the present invention is used, or the orientation or the positional relationship which is usually understood by those skilled in the art, or the orientation or the positional relationship which is usually placed when the product of the present invention is used, and is only for the convenience of describing the present invention and simplifying the description, but does not indicate or imply that the indicated device or element must have a specific orientation, be constructed in a specific orientation, and be operated, and therefore, cannot be understood as limiting the present invention. Furthermore, the terms "first" and "second" are used merely to distinguish one description from another, and are not to be construed as indicating or implying relative importance.
In the description of the embodiments of the present invention, it should be further noted that the terms "disposed" and "connected" are to be interpreted broadly, and may be, for example, fixedly connected, detachably connected, or integrally connected, unless explicitly stated or limited otherwise; may be directly connected or indirectly connected through an intermediate. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations. The drawings in the embodiments clearly and completely describe the technical solutions in the embodiments of the present invention, and obviously, the described embodiments are a part of the embodiments of the present invention, but not all of the embodiments.
As shown in fig. 1, 2 and 3, the present embodiment provides a device for detecting the sterilization effect of a disinfectant, comprising a base case 1 and a top cover 2 adapted to the base case 1, wherein the base case 1 is open at the top, and the top cover 2 covers the top of the base case 1 to close the base case 1. Be equipped with a plurality of cultivation check 3 that are used for placing the culture medium in the base box 1, top cap 2 is equipped with a plurality of dilution check 4, and dilution check 4 sets up in top cap 2's inboard, and the splendid attire diluent is convenient for dilute the sample in the dilution check 4. The culture lattice 3 may be circular or rectangular, and in the present embodiment, both the culture lattice 3 and the dilution lattice 4 are rectangular, and the size of the culture lattice 3 is the same as that of the dilution lattice 4.
The detection device further comprises an inoculator, wherein the inoculator is used for inoculating the sample of the diluted grid 4 into the culture medium for culturing the grid 3, the sample contains bacterial liquid, the bacterial liquid is cultured on the culture medium to grow bacterial colonies, and the results are compared according to the number of the bacterial colonies before and after the disinfectant is disinfected, so that the disinfection effect of the disinfectant is evaluated.
In one embodiment, the inoculator comprises an inoculation plate 5, on which inoculation plate 5 sets of inoculation needles 6 are provided. Each group of inoculating needles 6 consists of a plurality of inoculating needles 6, and each group of inoculating needles 6 is dipped with a sample of the dilution grid 4 and then inoculated into a culture medium of the culture grid 3. The inoculation plate 5 is slightly larger than the base box 1 and the top cover 2, when the inoculation plate 5 is covered on the top cover 2, the inoculation needle 6 is contacted with the sample, then the inoculation plate 5 is covered on the base box 1, and the inoculation needle 6 which is stained with the sample is contacted with the surface of the culture medium, thereby inoculating bacteria on the culture medium. The inoculation plate 5 can be provided with a handle, so that the inoculation plate 5 can be conveniently taken.
In one embodiment, the base case 1 has a length of 127mm, a width of 85mm, and a height of 23mm, and is provided with 3 rows and 4 columns of cultivation lattices 3, the volume of the culture medium in each cultivation lattice 3 is 3mL, the top cover 2 is provided with 3 rows and 4 columns of dilution lattices 4, and the cultivation lattices 3 and the dilution lattices 4 have the same size. In one embodiment, the medium composition of each row of culture grids 3 in the medium box 1 is different, the 3 rows of culture mediums are CN medium, BP medium and EMB medium respectively, the 3 rows of culture mediums are used for qualitatively analyzing microorganisms in the samples by using pseudomonas aeruginosa, staphylococcus aureus and escherichia coli, and the four grids of each row are used for counting samples with different concentrations respectively for quantitatively analyzing the microorganisms. The components of the culture medium in the medium box 1 can be set to be identical culture medium, which can be set according to actual requirements.
The CN culture medium is a pseudomonas agar basic culture medium and is used for detecting pseudomonas aeruginosa, and the gelatin peptone and the tryptone in the CN culture medium provide a nitrogen source; glycerol provides a carbon source; potassium sulfate and magnesium chloride can promote generation of pyocyanin; agar is the coagulant of the culture medium, nalidixic acid inhibits gram-negative bacilli other than pseudomonas. The pseudomonas aeruginosa shows green fluorescence on a culture medium plate, green pigment is generated at the dense position of the bacterial colony, and the fluorescence can be seen at the sparse position.
The BP culture medium is a culture medium and a Belde-Pack agar culture medium, the BP culture medium is used for detecting staphylococcus aureus, tellurite enriched yolk liquid is contained in the BP culture medium, the staphylococcus aureus can reduce tellurium elements in potassium tellurite, and the color of the staphylococcus aureus is brownish black or black. Staphylococcus aureus also produces lecithinase, which dissolves the yolk of the medium, forming a clear ring. The golden yellow colony is brownish black, and a circle of transparent ring is arranged around the golden yellow colony.
The EMB culture medium is eosin methylene blue agar culture medium, and is used for detecting escherichia coli. The EMB culture medium contains two indicators of eosin and methylene blue, and inhibits the growth of most gram-positive bacteria. The escherichia coli can ferment lactose to produce acid to cause an acid environment, so that the two indicators are combined to form a compound, and escherichia coli bacterial colonies are dark green with metal luster and are distinguished from other microorganisms which cannot ferment lactose to produce acid. Coli presents a purplish red colony with a black center with metallic luster.
In one embodiment, 3 groups of inoculating needles 6 arranged side by side are arranged on the inoculating plate 5, the length of the inoculating plate 5 is slightly larger than the width of the base box 1 and the top cover 2, and the inoculating needles 6 can inoculate samples with the same dilution concentration into three culture media at one time during inoculation, so that the operation is convenient, and the time and the labor are saved.
In one embodiment, 4 groups of inoculating needles 6 are arranged side by side on the inoculating plate 5, the length of the inoculating plate 5 is slightly longer than that of the base box 1 and the top cover 2, and the inoculating needles 6 can inoculate samples with different dilution concentrations into a culture medium at one time during inoculation, so that the operation is convenient, and the time and the labor are saved.
Each group of inoculating needles 6 comprises 20 inoculating needles 6 in 4 rows and 5 rows, so that the inoculating needles 6 can successfully complete inoculation, the distance between every two inoculating needles 6 is equal, the inoculating needles 6 are in a hollow conical shape, the length of each inoculating needle 6 is 8mm, and the diameter of one end of each inoculating needle, which is far away from the needle point, is 0.6 mm. The inoculation plates 5 can be overlapped, and the packaging space is saved.
The invention designs a novel detection device with disinfectant disinfection effect by improving the traditional detection device, and can simultaneously and qualitatively detect pseudomonas aeruginosa, escherichia coli and staphylococcus aureus. The components in the culture medium in the culture grid 3 of the detection device can effectively inhibit the growth of the mould and has higher selectivity on three detected target strains. The dilution grid 4 is used for detecting samples with different dilutions and quantitatively detecting microorganisms. The design of upper cover is used for diluting the sample in the detection device and uses, practices thrift the cost, need not independent dilution sample container, makes things convenient for the field usage in plant. The design of the inoculating needle 6 on the inoculator can quickly inoculate the sample and save the packaging space. The detection device can be used for rapidly, qualitatively and quantitatively detecting microorganisms and conveniently and rapidly evaluating the disinfection effect of the disinfectant.
The embodiment also provides a rapid evaluation method for the disinfection effect of a disinfectant, which comprises the detection device, and the rapid evaluation method comprises the following steps:
collecting samples: collecting samples before and after disinfection by a disinfectant;
diluting: the dilution chamber 4 in the top cover 2 contains PBS buffer, and each sample is diluted with the PBS buffer in accordance with a concentration gradient. The samples before and after sterilization are respectively used by a top cover 2, 3 rows and 4 columns of dilution grids 4 are arranged in the top cover 2, and the samples can be arranged according to 10 by the 4 columns of dilution grids 4-13 times with a concentration gradient.
Inoculation: inoculating each diluted sample into a culture medium in a base box 1 by using an inoculator, wherein 3 rows and 4 columns of cultivation grids 3 are arranged in the base box 1, the components of the culture medium of each row of cultivation grids 3 are different, and the 3 rows of culture media are respectively a CN culture medium, a BP culture medium and an EMB culture medium and are respectively used for detecting pseudomonas aeruginosa, staphylococcus aureus and escherichia coli. The inoculation plate 5 is provided with 3 groups of inoculation needles 6, and the inoculation needles 6 can inoculate samples with the same dilution concentration into three culture media at one time during inoculation, so that the operation is convenient, and the time and the labor are saved.
Culturing: culturing in 37 deg.C incubator for 12-24 hr;
and (5) counting results: and counting the number of bacterial colonies of each culture medium, and evaluating the disinfection effect of the disinfectant according to the comparison result of the number of bacterial colonies before and after disinfection of the disinfectant.
As shown in fig. 4, in one embodiment, the sample collection comprises the following processes: if the sample is solid, the sample is firstly sucked and washed by PBS buffer solution, then washing liquid is collected, sediment is collected after centrifugation, and the sediment is dispersed in the PBS buffer solution again to obtain the sample before and after disinfection by the disinfectant; and if the sample is liquid, performing concentration treatment according to the turbidity degree of the sample to obtain the sample before and after disinfection by the disinfectant. For example, before and after a field fecal sample is disinfected by a disinfectant, 1g of the sample is respectively taken, 6mL of PBS buffer solution is added, the sample is uniformly shaken and mixed, 2500g of supernatant is obtained by centrifugation, 12000g of supernatant is obtained by centrifugation for 10min, precipitate is obtained by centrifugation and dissolved in 1mLPBS for re-suspension, dilution is carried out by 2 times, 10 times and 100 times, and then the dilution is taken by using an inoculator for inoculation, and the detection result is shown in figure 5.
In one embodiment, the seeding comprises the following processes: each diluted sample is dipped by an inoculator and inoculated into the culture medium in the base box 1, and each diluted sample is subjected to three repeated samples, so that the influence of the randomness of the detection on the result is avoided.
The evaluation method can be used for rapidly detecting the disinfection effect of the disinfectant, and has the following advantages:
(1) the method has strong pertinence, and can simultaneously detect 3 typical microorganisms in disinfection effect evaluation, namely pseudomonas aeruginosa, escherichia coli and staphylococcus aureus;
(2) the method has the advantages that the method is precise and standard, the accuracy is high, dead cells killed by the disinfectant cannot grow on a flat plate, and living cells can be detected qualitatively; the component selectivity of the culture medium is strong, the target strains can be directly screened, and the non-target strains cannot grow on the flat plate;
(3) the operation is simple and rapid, qualitative and quantitative detection of microorganisms can be rapidly carried out through the evaluation method, and the disinfection effect of the disinfectant can be simply and rapidly evaluated;
(4) the observation result is visual, the shapes, sizes and colors of different microorganisms on the culture medium are different, and the colony count can be counted through direct observation by naked eyes.
The invention is not limited to the above alternative embodiments, and any other various forms of products can be obtained by anyone in the light of the present invention, but any changes in shape or structure thereof, which fall within the scope of the present invention as defined in the claims, fall within the scope of the present invention.
Claims (10)
1. A detection device for the disinfection effect of a disinfectant comprises a base box (1) and a top cover (2) matched with the base box (1), and is characterized in that a plurality of culture lattices (3) for placing a culture medium are arranged in the base box (1), and a plurality of dilution lattices (4) for diluting a sample are arranged on the top cover (2); further comprising an inoculator for inoculating a sample of the dilution grid (4) into the culture medium of the incubation grid (3).
2. The device for detecting the disinfection effect of a disinfectant according to claim 1, wherein the inoculator comprises an inoculating plate (5), and a plurality of groups of inoculating needles (6) are arranged on the inoculating plate (5).
3. The device for detecting the disinfection effect of a disinfectant according to claim 2, wherein M rows and N columns of cultivation lattices (3) are arranged in the base box (1), M rows and N columns of dilution lattices (4) are arranged on the top cover (2), the cultivation lattices (3) and the dilution lattices (4) have the same size, and M or N groups of inoculation needles (6) arranged side by side are arranged on the inoculation plate (5).
4. The device for detecting the sterilization effect of a sterilization agent according to claim 3, wherein the composition of the culture medium in each row of the culture compartments (3) in the base case (1) is different.
5. The apparatus for detecting sterilization effect of disinfectant according to claim 4, wherein said culture medium comprises at least one of CN culture medium, BP culture medium and EMB culture medium.
6. The device for detecting the disinfection effect of a disinfectant according to claim 2, wherein each group of inoculation needles (6) comprises X rows and Y rows of inoculation needles (6), and each inoculation needle (6) is conical.
7. The device for detecting the disinfection effect of a disinfectant according to claim 6, wherein each inoculation needle (6) has a length of 8mm and an end thereof away from the needle tip has a diameter of 0.6 mm.
8. A rapid evaluation method of the disinfection effect of a disinfectant, comprising the test device according to any one of claims 1 to 7, comprising the steps of:
collecting samples: collecting samples before and after disinfection by a disinfectant;
diluting: PBS buffer solution is filled in a dilution grid (4) in the top cover (2), and each sample is diluted by the PBS buffer solution according to concentration gradient;
inoculation: inoculating each diluted sample into the culture medium in the medium box (1) by using an inoculator;
culturing: culturing in 37 deg.C incubator for 12-24 hr;
and (5) counting results: and counting the colony number of each culture medium, calculating an average value, and evaluating the disinfection effect of the disinfectant according to the comparison result of the colony numbers before and after disinfection of the disinfectant.
9. The method for rapidly evaluating the disinfection effect of a disinfectant according to claim 8, wherein the sample collection comprises the following processes: if the sample is solid, the sample is firstly sucked and washed by PBS buffer solution, then washing liquid is collected, sediment is collected after centrifugation, and the sediment is dispersed in the PBS buffer solution again to obtain the sample before and after disinfection by the disinfectant; if the sample is liquid, carrying out concentration treatment according to the turbidity degree of the sample to obtain the sample before and after disinfection by the disinfectant; if the sample is air, the sample is directly collected by adopting a natural sedimentation method, or air is pumped and filtered by utilizing an air sampler, bacteria and dust in the air are collected into PBS buffer solution after impacting the water surface, and then the sample is concentrated to obtain the sample before and after being disinfected by the disinfectant.
10. The method for rapidly evaluating the disinfection effect of a disinfectant according to claim 8, wherein said inoculation comprises the following processes: each diluted sample was dipped with an inoculator and inoculated into the medium in the cassette (1), and three replicates were made for each diluted sample.
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| CN202010261363.6A CN111321068A (en) | 2020-04-03 | 2020-04-03 | Detection device for disinfection effect of disinfectant and rapid evaluation method |
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| CN202010261363.6A CN111321068A (en) | 2020-04-03 | 2020-04-03 | Detection device for disinfection effect of disinfectant and rapid evaluation method |
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| CN112378902A (en) * | 2020-11-03 | 2021-02-19 | 华中农业大学 | Method for rapidly evaluating disinfection effect of antibacterial disinfectant |
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| CN112378902A (en) * | 2020-11-03 | 2021-02-19 | 华中农业大学 | Method for rapidly evaluating disinfection effect of antibacterial disinfectant |
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