Reagent plate for rapid drug sensitivity test
By designing a reagent plate for rapid drug sensitivity testing, using semi-enclosed bottom structure heating and physical isolation of protective film, the problem that traditional drug sensitivity plates cannot meet the rapid clinical diagnosis is solved, and the effect of accelerating drug sensitivity experiments and improving experimental accuracy is achieved.
Patent Information
- Application Number
- CN202510273663.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-10
- Publication Date
- 2025-04-25
- Estimated Expiration
- Not applicable · inactive patent
AI Technical Summary
Traditional drug sensitivity plates cannot meet the needs of rapid clinical diagnosis, and the experimental process is relatively slow, so it is impossible to quickly obtain drug sensitivity results.
A reagent plate for rapid drug sensitivity testing was designed, which was heated using a semi-enclosed bottom structure. The protective film prevented bacterial samples from spilling out through physical isolation, and facilitated needle sample suction operation through cross marks.
Through semi-enclosed bottom structure heating, the bacterial growth adjustment period is shortened and the drug sensitivity experiment is accelerated; the protective film reduces biosafety risks, maintains the stability of the culture medium, and improves the experimental accuracy; expands the plate hole capacity and improves the equipment detection sensitivity.
Smart Images
Figure CN119875788A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of drug sensitivity test reagent plates, and in particular to a reagent plate for rapid drug sensitivity test. Background Art
[0002] Traditional drug sensitivity plates are usually 96-well plates with standardized well design and stable materials. They are mostly made of plastics with stable chemical properties and certain transparency, such as polystyrene. It is suitable for the relatively slow traditional drug sensitivity test process, and the operation is relatively simple and versatile. Taking the broth dilution method as an example, a broth culture medium containing nutrients is added to the well, and then a gradient diluted drug solution and bacterial suspension are added. The bacterial growth and reproduction make the liquid turbid, and the bacterial growth at different drug concentrations is observed to determine the minimum inhibitory concentration. The agar diffusion method can also be used. After the bacterial agar solidifies in the well, a drug-containing paper sheet is placed on the surface, and the drug sensitivity is judged according to the size of the growth inhibition zone under the concentration gradient formed by drug diffusion. In the interactive scheme, the experimenter uses a pipette to accurately add the bacterial suspension and drug solution to the well, and then puts it into the incubator for cultivation. The incubator provides a suitable temperature, humidity and gas environment. During the culture period, the bacterial growth can be monitored by observing the degree of turbidity with the naked eye or by using an enzyme marker to detect the optical density. Finally, the positive control well and the negative control well are compared to determine the result.
[0003] However, in traditional drug sensitivity tests, the speed at which bacteria enter the logarithmic phase is not required, and the experimental process is relatively slow, and there is no need to work closely with rapid drug sensitivity test equipment. However, with the increasing demand for rapid diagnosis in clinical practice, it is difficult to meet the demand for rapid clinical diagnosis. Based on this, providing a reagent plate for rapid drug sensitivity testing has important practical significance. Summary of the invention
[0004] The purpose of the present invention is to provide a reagent plate for rapid drug sensitivity testing, aiming to solve the technical problem that traditional drug sensitivity plates in the prior art cannot meet the requirements of clinical rapid diagnosis.
[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:
[0006] The present invention provides a reagent plate for rapid drug sensitivity testing, comprising:
[0007] A plate hole body, wherein the plate hole body is a flat plate structure, and a plate hole is provided at the lower part thereof;
[0008] Plate holes, the plate holes are arranged in a plurality of rows and columns and are evenly arranged in rows and columns at the bottom of the plate hole body;
[0009] A protective film is circumferentially arranged on the top of each of the plate holes to seal the plate holes.
[0010] Furthermore, a cross cut is provided in the middle of each protective film to facilitate processing of samples in the plate holes.
[0011] Furthermore, the plate hole is hollow inside and open at the top, and the bottom is a U-shaped structure; the bottom of the plate hole is a semi-enclosed structure.
[0012] Furthermore, the plate hole body also includes:
[0013] A handle, which is arranged on one side of the plate hole body and is used to take the plate hole body;
[0014] The buckle is arranged on the other side of the plate hole body to facilitate the use of the reagent plate with other equipment; the buckle is connected to the plate hole body in an integral manner to form a stepped structure.
[0015] Furthermore, the plate holes may be arranged irregularly at the lower part of the plate hole body.
[0016] The present invention also provides an application method of the reagent plate for rapid drug sensitivity testing described in the above technical solution, characterized in that it comprises the following steps:
[0017] Incubate the standard strain to obtain the bacteria to be tested;
[0018] The bacteria to be tested are mixed with AST broth, and then the bacteria to be tested are inoculated after mixing to obtain a bacterial solution;
[0019] The bacterial solution is added to the reagent plate and then incubated, the reagent plate is heated, and the growth of bacteria is observed.
[0020] Furthermore, the protective film of the reagent plate is made of polyethylene or polypropylene film material.
[0021] Furthermore, the heating is heating the bottom of the wells of the reagent plate.
[0022] Furthermore, the plate well capacity of the reagent plate is 500 mL.
[0023] Furthermore, the bacterial solution can be designed to have different concentration gradients and tested using the reagent plate for rapid drug sensitivity testing.
[0024] Compared with the prior art, the technical solution of the present invention has the following beneficial effects:
[0025] (1) The bottom of the plate hole of the present invention is a semi-enclosed structure. Based on the principle of heat conduction, the semi-enclosed bottom structure can conduct heat to the culture medium and bacterial sample in the hole in multiple directions when heated by the hot melt pool, thereby increasing the heat exchange area and efficiency, shortening the bacterial growth adjustment period, and accelerating the start of the drug sensitivity test;
[0026] (2) The present invention provides a protective film around the top of each plate hole, and a cross cut is provided in the middle of each protective film. The protective film prevents the bacterial sample from overflowing in a physical isolation manner, thereby reducing the biosafety risk. It can also prevent water evaporation, and by reducing the contact area between liquid and air, maintain the stability of the osmotic pressure and nutrient components of the culture medium, thereby ensuring the accuracy of the experiment. In addition, the cross cut facilitates the needle aspiration operation while ensuring the protective function of the protective film.
[0027] (3) The present invention expands the plate hole capacity, which can provide sufficient reaction space for bacteria, making the impedance change caused by bacterial metabolism more significant, and improving the detection sensitivity of the equipment based on the impedance principle. BRIEF DESCRIPTION OF THE DRAWINGS
[0028] Figure 1 is a schematic diagram of the reagent plate of the present invention;
[0029] Figure 2 is a side view of the reagent plate of the present invention;
[0030] Figure 3 A top view of the reagent plate of the present invention;
[0031] 1-plate hole body, 2-plate hole, 3-protective film, 4-handle, 5-cross cut, 6-buckle, 7-semi-enclosed structure. DETAILED DESCRIPTION
[0032] Traditional drug sensitivity plates are usually 96-well plates with standardized well design and stable materials. They are mostly made of plastics with stable chemical properties and certain transparency, such as polystyrene. It is suitable for the relatively slow traditional drug sensitivity test process, and the operation is relatively simple and versatile. Taking the broth dilution method as an example, a broth culture medium containing nutrients is added to the well, and then a gradient diluted drug solution and bacterial suspension are added. The bacterial growth and reproduction make the liquid turbid, and the bacterial growth at different drug concentrations is observed to determine the minimum inhibitory concentration. The agar diffusion method can also be used. After the bacterial agar solidifies in the well, a drug-containing paper sheet is placed on the surface, and the drug sensitivity is judged according to the size of the growth inhibition zone under the concentration gradient formed by drug diffusion. In the interactive scheme, the experimenter uses a pipette to accurately add the bacterial suspension and drug solution to the well, and then puts it into the incubator for cultivation. The incubator provides a suitable temperature, humidity and gas environment. During the culture period, the bacterial growth can be monitored by observing the degree of turbidity with the naked eye or by using an enzyme marker to detect the optical density. Finally, the positive control well and the negative control well are compared to determine the result.
[0033] However, in traditional drug sensitivity tests, the speed at which bacteria enter the logarithmic phase is not required, and the experimental process is relatively slow, and there is no need to work closely with rapid drug sensitivity test equipment. However, with the increasing demand for rapid diagnosis in clinical practice, it is difficult to meet the demand for rapid clinical diagnosis. Based on this, providing a reagent plate for rapid drug sensitivity testing has important practical significance.
[0034] The specific implementation of the present invention is further described in detail below in conjunction with the accompanying drawings and examples. The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.
[0035] In the description of the present application, it should be understood that the terms "center", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", etc., indicating orientations or positional relationships, are based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as a limitation on the present application.
[0036] The terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of the features. In the description of this application, unless otherwise specified, "plurality" means two or more.
[0037] In the description of this application, it should be noted that, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in this application can be understood according to specific circumstances.
[0038] The present invention provides a reagent plate for rapid drug sensitivity testing, comprising:
[0039] A plate hole body 1, wherein the plate hole body 1 is a flat plate structure, and a plate hole 2 is provided at the lower part thereof;
[0040] Plate holes 2, the plate holes 2 are arranged in a plurality of rows and columns, and are evenly arranged in rows and columns at the bottom of the plate hole body 1;
[0041] The protective membrane 3 is circumferentially arranged on the top of each of the plate holes 2 to seal the plate holes 2 .
[0042] In the present invention, a cross cut 5 is provided in the middle of each of the protective films 3 to facilitate processing of the samples in the plate holes.
[0043] In the present invention, the plate hole 2 is hollow inside and open at the top, and the bottom is a U-shaped structure; the bottom of the plate hole 2 is a semi-enclosed structure 7.
[0044] In the present invention, the plate hole body 1 further includes:
[0045] A handle 4, which is disposed on one side of the plate hole body 1 and is used to take the plate hole body 1;
[0046] The buckle 6 is arranged on the other side of the plate hole body 1 to facilitate the use of the reagent plate with other equipment; the buckle 6 is connected to the plate hole body as a whole and presents a stepped structure.
[0047] In the present invention, the plate holes 2 may be arranged irregularly at the lower part of the plate hole body 1 .
[0048] The present invention also provides an application method of the reagent plate for rapid drug sensitivity testing described in the above technical solution, characterized in that it comprises the following steps:
[0049] Incubate the standard strain to obtain the bacteria to be tested;
[0050] The bacteria to be tested are mixed with AST broth, and then the bacteria to be tested are inoculated after mixing to obtain a bacterial solution;
[0051] The bacterial solution is added to the reagent plate and then incubated, the reagent plate is heated, and the growth of bacteria is observed.
[0052] In the present invention, the standard strain is preferably selected according to actual application and then subjected to drug sensitivity test. The present invention does not specifically limit the mixing ratio of the test bacteria and AST broth and the incubation conditions of the bacterial solution. The mixing ratio and incubation conditions familiar to those skilled in the art can be adopted according to the changes in the strains.
[0053] In the present invention, the protective film of the reagent plate is preferably made of polyethylene or polypropylene film material.
[0054] In the present invention, the heating is preferably performed to heat the bottom of the plate well 2 of the reagent plate.
[0055] In the present invention, the plate well capacity of the reagent plate is preferably 500 mL.
[0056] In the present invention, the bacterial solution can be designed to have different concentration gradients and tested using the reagent plate for rapid drug sensitivity testing.
[0057] In the present invention, unless otherwise specified, the required raw materials for preparation are all commercially available products well known to those skilled in the art.
[0058] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.
[0059] Application Example 1
[0060] (1) Preparation of bacterial strains: Three standard strains (ATCC 25922 Escherichia coli, ATCC 25923 Staphylococcus aureus, ATCC 27853 Pseudomonas aeruginosa) were transferred for use and incubated at 37°C for 18 hours;
[0061] (2) Broth preparation: Grind the prepared strain on the wall of the AST (antibiotic susceptibility test) broth bottle, mix well, cover the bottle cap and measure the turbidity with a turbidimeter (BD PhoenixSpec Nephelomter). The turbidity is 0.5 McFarland units and set aside. Inoculate the bacteria to be tested according to the inoculation concentration of the broth dilution method of the CLSI (Clinical Laboratory Standards Institute);
[0062] (3) Temperature change: The prepared bacterial solution was added to the drug-sensitive plate of the present invention and placed in FASTCHECK for incubation; the prepared bacterial solution was added to an ordinary 96-well plate and placed in a constant temperature incubator, the temperature was set to 37°C, and temperature detectors were added to the two plates respectively, and the temperature of the two drug-sensitive plates was recorded within 3 hours. The results are shown in Table 1:
[0063] Table 1 Analysis of temperature changes in broth culture of drug sensitivity plate
[0064]
[0065] Based on Table 1, it can be seen that the rapid drug sensitivity plate maintains a stable temperature of 36°C-37°C from 4 minutes to 180 minutes, and it takes about 180 minutes for an ordinary 96-well plate to reach 36°C-37°C. Therefore, the temperature inside the reagent plate for rapid drug sensitivity testing described in the present invention rises rapidly, which can speed up the progress of the drug sensitivity test.
[0066] Application Example 2
[0067] 500ul of physiological saline was added to two drug sensitivity plates respectively, and FASTCHECK was placed. Plate 1 was covered with a sealing film, and plate 2 was not covered with a film. After incubation for 2 hours, the mass of the remaining solution in the plate was recorded with a balance. The results are shown in Table 2.
[0068] Table 2 Analysis of the effect of the sealing film on the solution volume change during the two-hour incubation of the drug sensitivity plate
[0069]
[0070] Based on Table 2, it can be seen that with the sealing film, the mass of the solution decreased by 0-0.2% after 2 hours of incubation, and the mass of the solution without the sealing film decreased by 20.22-38.20%. The protective film in the present invention greatly avoids the increase in concentration caused by water evaporation, which causes the error problem of drug sensitivity data, and improves the accuracy rate on the basis of realizing rapid drug sensitivity testing.
[0071] Application Example 3
[0072] (1) Preparation of bacterial strains: Three standard strains (ATCC 25922 Escherichia coli, ATCC 25923 Staphylococcus aureus, ATCC 27853 Pseudomonas aeruginosa) were transferred for use and incubated at 37°C for 18 hours;
[0073] (2) Broth preparation: Grind the prepared strain on the wall of the AST (antibiotic susceptibility test) broth bottle, mix well, cover the bottle cap and measure the turbidity with a turbidimeter (BD PhoenixSpec Nephelomter). The turbidity is 0.5 McFarland units and set aside. Inoculate the bacteria to be tested according to the inoculation concentration of the broth dilution method of the CLSI (Clinical Laboratory Standards Institute);
[0074] (3) The antibiotics on the whole page are randomly arranged in order, as shown in Table 3, and compared with the drug sensitivity plates 1, 2, and 3 arranged in the present invention, as shown in Table 4, to conduct a drug sensitivity test to test the accuracy of the drug sensitivity results.
[0075] Table 3 Random arrangement of antibiotics on rapid drug sensitivity plates
[0076]
[0077]
[0078] Table 4 Antibiotic drug arrangement table of rapid drug sensitivity plate
[0079]
[0080]
[0081]
[0082]
[0083] The experimental results based on Table 3 and Table 4 are shown in Table 5 and Table 6;
[0084] Table 5 ATCC 25922 Escherichia coli drug sensitivity test results
[0085]
[0086]
[0087] Table 6 ATCC 25923 Staphylococcus aureus drug sensitivity test results
[0088]
[0089]
[0090] Based on Tables 3 to 6, it can be seen that the quality control result conformity rate of the randomly arranged drug sensitivity plate of the ATCC 25922 Escherichia coli drug sensitivity test is 42%, and the quality control result conformity rate of the rapid drug sensitivity plate of the present invention is 100%; the quality control result conformity rate of the randomly arranged drug sensitivity plate of the ATCC 25923 Staphylococcus aureus drug sensitivity test is 63%, and the quality control result conformity rate of the rapid drug sensitivity plate of the present invention is 100%.
[0091] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.
Claims
1. A reagent plate for rapid drug sensitivity testing, characterized in that: include: A plate hole body, wherein the plate hole body is a flat plate structure, and a plate hole is provided at the lower part thereof; Plate holes, the plate holes are arranged in a plurality of rows and columns and are evenly arranged in rows and columns at the bottom of the plate hole body; A protective film is circumferentially arranged on the top of each of the plate holes to seal the plate holes.
2. The reagent plate for rapid drug sensitivity testing according to claim 1, characterized in that: A cross cut is arranged in the middle of each protective film to facilitate processing of the sample in the plate hole.
3. The reagent plate for rapid drug sensitivity testing according to claim 1, characterized in that: The plate hole is hollow inside and open at the top, and the bottom is a U-shaped structure; the bottom of the plate hole is a semi-enclosed structure.
4. The reagent plate for rapid drug sensitivity testing according to claim 1, characterized in that: The plate hole body also includes: A handle, the handle being arranged on one side of the plate hole body and being used for taking the plate hole body; The buckle is arranged on the other side of the plate hole body to facilitate the use of the reagent plate with other equipment. The buckle is connected to the plate hole body as a whole and presents a stepped structure.
5. The reagent plate for rapid drug sensitivity testing according to claim 1, characterized in that: The plate holes may be arranged irregularly at the lower part of the plate hole body.
6. A method for using the reagent plate for rapid drug sensitivity testing according to any one of claims 1 to 5, characterized in that: The following steps are involved: Incubate the standard strain to obtain the bacteria to be tested; The bacteria to be tested are mixed with AST broth, and then the bacteria to be tested are inoculated after mixing to obtain a bacterial solution; The bacterial solution is added to the reagent plate and then incubated, the reagent plate is heated, and the growth of bacteria is observed.
7. The method for using the reagent plate for rapid drug sensitivity testing according to claim 6, characterized in that: The protective film of the reagent plate is made of polyethylene or polypropylene film material.
8. The method for using the reagent plate for rapid drug sensitivity testing according to claim 6, characterized in that: The heating is heating the bottom of the plate wells of the reagent plate.
9. The method for using the reagent plate for rapid drug sensitivity testing according to claim 6, characterized in that: The plate well capacity of the reagent plate is 500 mL.
10. The method for using the reagent plate for rapid drug sensitivity testing according to claim 6, characterized in that: The bacterial solution can be designed to have different concentration gradients and tested using the reagent plate for rapid drug sensitivity testing.