CN113845798A

CN113845798A - Antibacterial drug ink and method for preparing ester drug sensitive test strip by using same

Info

Publication number: CN113845798A
Application number: CN202111081064.5A
Authority: CN
Inventors: 吴秀祯; 王志贤; 刘昶; 盛长忠; 粟艳; 周泽奇
Original assignee: Dynamiker Biotechnology Tianjin Co Ltd
Current assignee: Dynamiker Biotechnology Tianjin Co Ltd
Priority date: 2021-09-15
Filing date: 2021-09-15
Publication date: 2021-12-28

Abstract

The invention provides an antibacterial drug ink and a method for preparing an ester drug sensitive test strip by using the same. The antibacterial medicine ink comprises the following components in percentage by mass: 0.01 to 10 percent of antibacterial drug, 0.5 to 5 percent of humectant, 0.5 to 2 percent of surfactant and the balance of deionized water. And injecting the antibacterial drug ink into an ink box of an ink-jet printer, carrying out ink-jet printing on a matrix material of the Etest drug sensitive test strip, drying and sterilizing to obtain the Etest drug sensitive test strip. The preparation method is simple in process and suitable for industrial production, and the Etest drug sensitive test strip prepared by the method is sensitive and accurate in detection result and high in reliability.

Description

Antibacterial drug ink and method for preparing ester drug sensitive test strip by using same

Technical Field

The invention belongs to the technical field of in-vitro diagnostic reagents for testing medicine, particularly relates to a preparation method of an Etest drug sensitive test strip, and particularly relates to antibacterial drug ink and a method for preparing the Etest drug sensitive test strip by using the same.

Background

With the wide application of broad-spectrum antibacterial drugs, the problem of drug resistance of the antibacterial drugs becomes more and more serious in clinic, and the problem of drug resistance of the antibacterial drugs is urgently solved. The drug susceptibility test (AST) is an important means for guiding clinicians to select antibiotics and inhibit the drug resistance of antibacterial drugs. At present, widely accepted standardization methods for antimicrobial drug susceptibility tests at home and abroad are mainly the dilution and diffusion methods recommended by the American Clinical Laboratory Standards Institute (CLSI).

Dilution-based susceptibility testing can be used to quantitatively determine the in vitro activity of an antibacterial agent against a particular bacterium and is classified into agar dilution and broth dilution. In an experiment, the concentration of an antibacterial drug is usually diluted by multiple times, the lowest drug concentration capable of inhibiting the visible growth of the flesh and eyes of a bacterium to be tested is called as the Minimum Inhibitory Concentration (MIC), and the test concentration range of a specific antibacterial drug should contain the concentration of an explanatory break point (sensitive, intermediate and drug-resistant) capable of detecting the bacterium and also should contain the MIC of a quality control reference strain. The dilution method is accurate and has wide application range, but has complex operation, time and labor waste and is not beneficial to clinical use.

The paper diffusion method is that paper containing quantitative antibacterial drugs is pasted on an agar plate inoculated with bacteria to be detected, the drugs contained in the paper absorb water in the agar and are continuously diffused to the area around the paper after being dissolved to form a decreasing gradient concentration, and the growth of the bacteria to be detected is inhibited within the range of the bacteriostatic concentration around the paper, so that a transparent bacteriostatic zone is generated. The size of the inhibition zone reflects the sensitivity of the detection bacteria to the determination drug, and is inversely related to the MIC of the drug to the bacteria to be detected, namely the larger the inhibition zone is, the smaller the MIC is. The paper diffusion method is a semi-quantitative drug sensitive detection method, is simple and easy to implement, cannot give an intuitive MIC value, and is influenced by various factors.

In order to overcome the defects of the two drug sensitive detection methods, the abbiometer test, sweden, has been introduced by abbioodisk, inc, to accurately determine the minimum inhibitory concentration of an antibacterial drug, and has been a reference standard in many microbiological laboratories, with convenient and rapid use, and can detect more than 100 antibiotics. The test strip for testing Etest is a plastic strip with the specification of 5 x 50mm, and one surface of the test strip is adsorbed with Log₂On the basis of the continuously changing antibiotic concentration, the other surface is marked with a corresponding antibiotic concentration scale. When the test strip of test is placed on a specific drug-sensitive medium inoculated with a certain concentration of bacteria, the antibiotic is immediately released into the agar and diffuses in a single pass. An oval bacteriostatic loop may be produced after incubation at 35 ℃ for a period of time. The scale at the junction of the antibacterial ring and the test strip is the MIC value aiming at the strain.

The concentration gradient test paper method is a great revolution of a drug sensitivity test method, combines the advantages of a diffusion method and a dilution method, simultaneously makes up for some defects of the diffusion method and the dilution method, can directly and quantitatively measure the MIC value of the antibacterial drug to the tested bacteria like the dilution method, and has accurate result and good repeatability. The concentration gradient test paper method can be used for drug sensitivity tests of various common bacteria, aerobic bacteria, mycobacteria, anaerobic bacteria, fungi and the like. Compared with the traditional paper diffusion method, the method for measuring the size of the inhibition zone is accurate and reliable.

At present, the Etest detection test strip is complex in production method and is not easy to produce in a large scale. For example, CN1667414A discloses a stepped concentration test strip and a preparation method thereof, which adopts a silk-screen printing technique to produce an ester drug sensitive test strip. The method is expensive in equipment, and one equipment can only produce a drug sensitive test strip of an antibacterial drug. CN2194001Y discloses an antibacterial drug sensitive gradient test strip, wherein 15 pieces of small paper containing different concentrations are adhered on a 5 x 50mm polyester sheet by pressure sensitive adhesive according to the concentration sequence, the production process is complex and difficult to standardize, and the quality of the product cannot be ensured. CN2462387Y discloses a concentration gradient drug sensitive strip, which is coated with antibiotic according to continuous concentration gradient and marked with the MIC scale. That is, the antibiotic concentration is the same on any line in the transverse direction, and the antibiotic is continuously and exponentially distributed on any line in the longitudinal direction. The drug distribution of the product of the technology is in a continuously stepless change concentration gradient instead of a step-type concentration gradient, and the current production equipment and technology can hardly carry out the production. CN112444624A discloses a method for manufacturing a drug sensitive test strip, which is too complicated to realize mass production.

In addition, CN205484363U discloses a small-sized concentration gradient drug sensitive test paper, which is printed or pasted with 15 antibacterial drugs with two times dilution concentration range on a 5 × 25mm plastic strip. The length of the paper strips is shortened by one time, the scales are reduced by half, more antibiotic paper strips can be placed on a culture medium with the same diameter, and innovation and improvement are not carried out on the production process.

Although the concentration gradient test strip method is approved by most industry standards at present, the Etest drug sensitive test strip is relatively complex in preparation method, so that the Etest drug sensitive test strip is expensive and difficult to be widely used clinically.

Therefore, the development of a new Etest drug sensitive paper strip production process has important values for reducing the detection cost, improving the clinical detection accuracy and guiding the reasonable utilization of antibiotics.

Disclosure of Invention

Aiming at the defects in the prior art, the invention aims to provide antibacterial drug ink and a method for preparing an Etest drug sensitive test strip by using the antibacterial drug ink. The antibacterial drug ink can be well adhered to a matrix material of a test strip, and when the antibacterial drug ink contacts a drug sensitive culture medium, the antibacterial drug contained in the test strip can also be normally diffused, so that the antibiotic concentration scale mark of the drug sensitive test strip is consistent with the actual drug diffusion condition, and the obtained detection result is accurate and reliable.

In order to achieve the purpose, the invention adopts the following technical scheme:

in a first aspect, the present invention provides an antibacterial drug ink for preparing an ester drug sensitive test strip, the antibacterial drug ink comprising, by mass: 0.01 to 10 percent of antibacterial drug, 0.5 to 5 percent of humectant, 0.5 to 2 percent of surfactant and the balance of deionized water.

The invention provides an antibacterial drug ink for preparing an ester drug sensitive test strip, which comprises an antibacterial drug, deionized water, a humectant and a surfactant. The humectant can adjust the drying speed of the ink, so that the ink is continuous or lacks strokes, and the surfactant enables the ink to wet the surface of paper more easily, thereby improving the printing effect on the paper; the humectant and the surfactant are matched with each other, so that the medicine can be better adhered to the matrix material of the medicine sensitive test strip, and the medicine can be better dispersed when contacting the medicine sensitive culture medium; the antibacterial medicine ink is high in drying speed in the printing process, and no precipitate is generated in the ink; the drug sensitive test strip prepared from the antibacterial drug ink has accurate drug gradient and good long-term storage stability.

In the present invention, the mass fraction of the antibacterial agent may be 0.01% to 10%, for example, 0.01%, 0.05%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, or the like.

In the invention, the mass fraction of the antibacterial drug can be specifically selected according to the detection concentration range of the antibacterial drug.

In the present invention, the humectant may be 0.5% to 5% by mass, for example, 0.5%, 0.8%, 1%, 1.2%, 1.4%, 1.5%, 1.8%, 2%, 2.5%, 2.8%, 3%, 3.2%, 3.5%, 3.8%, 4%, 4.5%, 4.8%, or 5%.

In the present invention, the surfactant may be present in an amount of 0.5% to 2% by mass, for example, 0.5%, 0.6%, 0.8%, 1%, 1.2%, 1.4%, 1.5%, 1.8%, or 2%.

In the invention, the antibacterial drug can be prepared into the antibacterial drug mother solution and then mixed with other components; the antibacterial drug can be dissolved in deionized water, organic solvent or buffer solutions such as PBS, HEPES-NaOH, Tris-HCl and the like.

Preferably, the organic solvent required for preparing the antibacterial mother liquor comprises ethanol, methanol, DMSO or the like, and the mass fraction of the organic solvent can be adjusted according to actual conditions, preferably 1 to 10%, and for example, may be 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, and the like.

In a preferred embodiment of the present invention, the antimicrobial ink further comprises 0.1 to 0.2M of a pH adjuster, which may be, for example, 0.1M, 0.12M, 0.14M, 0.15M, 0.16M, 0.17M, 0.18M, or 0.2M.

In the invention, the pH regulator provides proper pH value, thereby protecting an ink cavity and a spray head of the printer.

Preferably, the pH adjusting agent comprises any one or a combination of at least two of PBS, HEPES-NaOH or Tris-HCl, preferably Tris-HCl.

Preferably, the pH of the antibiotic ink is 7.0 to 8.0, and may be, for example, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.0.

As a preferred technical scheme of the invention, the humectant comprises 1, 3-butanediol and/or 1, 2-propanediol, and 1, 3-butanediol is preferred.

Preferably, the humectant is present in a mass fraction of 2% to 5%, and may be, for example, 2%, 2.2%, 2.5%, 2.8%, 3%, 3.2%, 3.4%, 3.5%, 3.8%, 4%, 4.2%, 4.5%, 4.6%, 4.8%, or 5%, etc., preferably 3%.

Preferably, the surfactant comprises sodium dodecylbenzene sulfonate.

Preferably, the surfactant is present in a mass fraction of 1% to 2%, and may be, for example, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, or the like, preferably 1.5%.

As a preferable technical scheme, the pH value of the antibacterial medicine ink is 7.0-8.0, and the antibacterial medicine ink comprises 0.01-10% by mass of antibacterial medicine, 2-5% by mass of 1, 3-butanediol, 1-2% by mass of sodium dodecyl benzene sulfonate, 0.1-0.2M of Tris-HCl and deionized water.

In a second aspect, the present invention provides the use of an antimicrobial ink according to the first aspect in the manufacture of an etet drug sensitive strip.

In a third aspect, the present invention provides a method for preparing an ester drug sensitive test strip based on an inkjet printing method, the method comprising the steps of:

(1) preparing the antibacterial drug ink according to the first aspect, and injecting the antibacterial drug ink into an ink cartridge of an inkjet printer;

(2) and printing the antibacterial drug ink on a matrix material of an Etest drug sensitive test strip by using the ink-jet printer, drying and sterilizing to obtain the Etest drug sensitive test strip.

The invention provides a method for preparing an ester drug sensitive test strip based on an ink jet printing technology, which is characterized in that different antibacterial drug mother solutions are dissolved in corresponding ink solvents and injected into an ink box of an ink jet printer, and the ink jet printer can select an ink jet printer capable of freely replacing ink jet materials. Printing different antibacterial drugs on the substrate material in a concentration gradient manner under the control of a computer, and drying and sterilizing the printed substrate material to obtain the antibacterial paper.

As a preferable technical scheme of the invention, the operation of filtering the antibacterial medicine ink is further included after the antibacterial medicine ink is prepared in the step (1).

Preferably, the filtration method is filtration by using a 0.22 μm filter membrane.

As a preferred technical solution of the present invention, the substrate material in step (2) includes any one or a combination of at least two of printing paper, qualitative filter paper, fiber film, polyacrylamide, polyester or polyamide, preferably printing paper.

The base material for spray printing selects A4 printing paper, is convenient to use, has moderate thickness and air permeability, avoids bubbles at the contact part of the culture medium and the test strip, and ensures the validity of the test result. If the substrate is a transparent plastic sheet, the prepared test strip is thin, the test strip is difficult to take out of the package due to strong electrostatic attraction between the transparent plastic sheet and the substrate, and meanwhile, the plastic sheet is airtight, and when the plastic sheet is applied to the surface of a solid culture medium, air bubbles are easily generated if the plastic sheet is not operated properly, so that the diffusion of the antibacterial drug in the drug sensitive test strip to the culture medium is influenced, and the test is invalid or inaccurate.

According to the invention, the antibacterial drug ink is combined with printing paper, and the prepared ester drug sensitive test strip is convenient to use and has air permeability, the antibacterial drug of the drug sensitive test strip is accurately diffused, and the test result is accurate and effective.

Preferably, the thickness of the printing paper is 0.03-0.2 mm, for example, 0.03mm, 0.05mm, 0.06mm, 0.08mm, 0.1mm, 0.12mm, 0.14mm, 0.15mm, 0.16mm, 0.18mm, 0.2mm, etc., preferably 0.1 mm.

Preferably, the drying in step (2) is specifically performed by: placing the substrate material containing the antibacterial ink in an environment with an air humidity less than 50%, and drying at 25-35 deg.C (such as 25 deg.C, 28 deg.C, 29 deg.C, 30 deg.C, 32 deg.C, 34 deg.C or 35 deg.C) for 1-5 min (such as 1min, 1.5min, 2min, 2.5min, 3min, 3.5min, 4min, 4.5min or 5 min).

As a preferred technical scheme of the invention, the sterilization mode of the step (2) is irradiation sterilization.

Preferably, the drying in the step (2) further comprises cutting and packaging operations.

Preferably, the packaging material of the test strip is an aluminum-plastic blister package.

As a preferred technical scheme of the invention, the method comprises the following steps:

(1) preparing an antimicrobial ink according to the first aspect, the antimicrobial ink comprising: 0.01-10% of antibacterial agent, 2-5% of 1, 3-butanediol, 1-2% of sodium dodecyl benzene sulfonate, 0.1-0.2M of Tris-HCl and deionized water;

filtering the antibacterial medicine ink by a filter membrane of 0.22 mu m, and injecting the antibacterial medicine ink into an ink box of an ink-jet printer;

(2) the antibacterial drug ink is printed on pre-printed printing paper with the thickness of 0.03-0.2 mm by using the ink-jet printer, the printing paper is placed in an environment with the air humidity of less than 50%, the printing paper is dried for 1-5 min at the temperature of 25-35 ℃, the printing paper is cut, an aluminum-plastic bubble cap is used for packaging, and the ester drug sensitive test strip is obtained after irradiation sterilization.

The recitation of numerical ranges herein includes not only the above-recited values, but also any values between any of the above-recited numerical ranges not recited, and for brevity and clarity, is not intended to be exhaustive of the specific values encompassed within the range.

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

(1) the invention provides an antibacterial drug ink, wherein the humectant and the surfactant in the antibacterial drug ink are not available, and the humectant and the surfactant are matched, so that the ink is easy to wet the surface of paper, and the antibacterial drug is easy to combine on a drug sensitive test strip, so that the printing effect on the paper is improved, and meanwhile, the drying speed of the ink can be adjusted, so that the ink is not broken or lacks strokes in the printing process;

(2) the method for preparing the Etest drug sensitive test strip by using the antibacterial drug ink is simple, the Etest drug sensitive test strip can be prepared by using the existing ink-jet printing technology, the preparation difficulty and the production cost of the Etest drug sensitive test strip are greatly reduced, and the method is suitable for industrial production.

Drawings

Fig. 1 is a schematic structural diagram of an Etest drug sensitive test strip prepared in application example 1, in which numbers in a box from top to bottom indicate the percentage of the antibacterial drug ink.

Detailed Description

The technical solutions of the present invention are further described in the following embodiments with reference to the drawings, but the following examples are only simple examples of the present invention and do not represent or limit the scope of the present invention, which is defined by the claims.

In the following examples, reagents and consumables used were obtained from conventional reagent manufacturers in the field unless otherwise specified; unless otherwise indicated, all experimental methods and technical means are conventional in the art.

Materials:

sodium dodecylbenzene sulfonate was purchased from Sigma-Aldrich;

fluconazole, amphotericin B and flucytosine were purchased from TCI echiei (shanghai) chemical industry development limited;

candida albicans standard strain ATCC90028 was purchased from ATCC.

Test example 1

The present test example tests the effect of the mass fraction of surfactant on the surface tension of an antibacterial drug ink.

And setting antibacterial medicine ink groups containing sodium dodecyl benzene sulfonate with different mass fractions. The antibacterial drug ink to be detected comprises 1% of fluconazole, 3% of 1, 3-butanediol, 0.1M Tris-HCl and deionized water in mass fraction, sodium dodecyl benzene sulfonate with corresponding mass fraction is respectively added, and the pH value is adjusted to 8.0. The surface tension of the antibiotic drug inks of different groups was measured, and the grouping and measurement results are shown in table 1.

TABLE 1

Sodium dodecylbenzenesulfonate (%)	0.1	0.5	1.0	1.5	2.0	2.5
							Surface tension (dyne)	49.34	41.26	35.24	31.32	25.43	20.46

The higher the proportion of surfactant in the solution, the lower the surface tension of the ink, because the hydrophilic part of the surfactant is dissolved in water and the lipophilic part extends to air, forming a transition layer on the air and water surfaces, lowering the surface tension of the ink. If the addition amount of the surfactant is small and the surface tension of the ink is too large, the printing effect is poor; if the amount of the surfactant added is large, the surface tension is lowered, but the stability of the ink is affected. Usually, the surface tension of the ink is controlled to be 32 to 36 dynes, and the detection results in table 1 show that the selectable mass fraction of the surfactant in the formula is 0.5 to 2%, and more preferably 1.0 to 1.5%.

Test example 2

This test example tests the effect of the mass fraction of humectant on the performance of an antibacterial ink.

Antibiotic drug ink groupings containing different mass fractions of 1, 3-butanediol were set. The antibacterial drug ink to be detected comprises 1% of fluconazole, 1.5% of sodium dodecyl benzene sulfonate, 0.1M Tris-HCl and deionized water in mass fraction, 1, 3-butanediol in corresponding mass fraction is added respectively, and the pH value is adjusted to 8.0. The performances of the antibiotic medicine inks of different groups are tested, and the grouping condition and the test result are shown in table 2.

TABLE 2

The viscosity and the drying speed of the ink have great relation with the addition amount of the humectant, and the proper addition amount can prevent the nozzle from being blocked by the ink. If the addition amount of the humectant is too small, the ink dries too quickly, so that the nozzle is easily blocked; if the amount of the humectant is too large, drying is not easy, and the accuracy of the drug concentration is affected to some extent. By combining the test results of the printing effect test in table 2, it can be seen that when the addition amount of the humectant is in the range of 0.5% to 5%, a better printing effect can be obtained, and when the addition amount of the humectant is in the range of 3% to 5%, the printing effect is more ideal.

Example 1

The embodiment provides an antibacterial drug ink for preparing an ester drug sensitive test strip;

the antibacterial medicine ink comprises the following components in percentage by mass: 1% fluconazole, 3% 1, 3-butanediol, 1.5% sodium dodecyl benzene sulfonate, 0.1M Tris-HCl and deionized water, and the pH value is 8.0.

Example 2

the antibacterial medicine ink comprises the following components in percentage by mass: 0.01% amphotericin B, 5% 1, 2-propanediol, 2% sodium dodecylbenzenesulfonate, 0.2M PBS and deionized water, pH 7.0.

Example 3

the antibacterial medicine ink comprises the following components in percentage by mass: 10% fluorocytosine, 0.5% 1, 3-butanediol, 0.5% sodium dodecylbenzenesulfonate, 0.15M HEPES-NaOH and deionized water, pH 8.0.

Example 4

the difference from example 1 is only that the antibiotic drug ink does not contain a pH adjusting agent, and the pH thereof is a naturally occurring pH value.

Example 5

the difference from example 1 is only that sodium dodecylbenzenesulfonate is replaced with sodium dodecylbenzenesulfonate in the antibacterial ink, and the remaining components are identical to those of example 1.

Example 6

the difference from example 1 is only that 1, 3-butanediol is replaced with glycerol in the antibiotic drug ink and the remaining components are in accordance with example 1.

Comparative example 1

The present comparative example provides an antimicrobial ink for the preparation of an Etest drug sensitive test strip;

the difference from example 1 is only that 1, 3-butanediol is not contained in the antibiotic drug ink and the contents of the remaining components are in accordance with example 1.

Comparative example 2

the difference from example 1 is only that the antibacterial ink does not contain sodium dodecylbenzenesulfonate and the contents of the remaining components are the same as those of example 1.

Comparative example 3

the difference from example 1 is only that 1, 3-butanediol was not contained in the antibiotic drug ink, the content of sodium dodecylbenzenesulfonate was increased to 4.2%, and the contents of the remaining components were maintained in accordance with example 1.

Comparative example 4

the difference from example 1 is only that the antibiotic drug ink does not contain sodium dodecylbenzenesulfonate, the content of 1, 3-butanediol is increased to 4.2%, and the contents of the remaining components are identical to example 1.

Comparative example 5

This comparative example provides a standard ink, which is the original ink for the printer.

Test of dehumidification speed of antibiotic drug ink

The drying time (sec) of one drop of ink (0.05mL) on a standard printing paper was measured to test the dewing speed, and the test results are shown in table 3.

TABLE 3

	Speed of dehumidification (second)
		Example 1	5
Example 2	5.5
		Example 3	4.5
Example 4	5
		Example 5	5
Example 6	4
		Comparative example 1	3
Comparative example 2	5
		Comparative example 3	3
Comparative example 4	7
		Comparative example 5	5

The test results in table 3 show that the content and kind of the moisture-absorbing agent in the ink affect the moisture-removing rate of the ink. If the dehumidification speed is too fast, the spray head is easy to block; if the dehumidification rate is too slow, the printed material is wet and not easily dried. Generally speaking, the dehumidification speed is preferably 4.5 to 5.5 seconds, and the dehumidification speeds of the antibacterial drug inks prepared in examples 1 to 5 and comparative example 2 are within a reasonable range, so that the antibacterial drug inks have higher use value;

in example 6, glycerin is used to replace 1, 3-butanediol, so that the dehumidification speed is increased, and the glycerin is selected to cause the blockage of the spray head more easily; the method has the advantages that 1, 3-butanediol is not added in the comparative example 1, 3-butanediol is not added in the comparative example 3, the 1, 3-butanediol which is lacked is complemented by sodium dodecyl benzene sulfonate, the spray head is easy to block due to too fast drying, and the printing effect and quality are influenced; comparative example 4 has a large amount of 1, 3-butanediol, a slow dehumidification rate, difficulty in drying, and may affect the drug concentration.

In conclusion, it can be seen that the dehumidification speed can be controlled within a proper range only by selecting 1, 3-butanediol and/or 1, 2-propanediol as the humectant and controlling the addition amount within the range of 0.5% -5%, and the printing effect cannot be influenced by the blockage of the nozzle, and the accuracy of the test strip cannot be influenced by slow drying.

Stability test of antibacterial ink

The unstable ink of the system can generate sedimentation phenomenon in the standing process. In the experiment, the ink of example 1 was poured into a graduated cylinder, and the interface between the precipitated liquid and the precipitated solid was tested for the downward movement, and compared with comparative example 5.

The inks of example 1 and comparative example 5 were examined for pH, viscosity, print smoothness, and stability, and the results are shown in table 4.

TABLE 4

Index (I)	pH	Viscosity of the oil	Fluency of printing	Stability of
					Example 1	8.01	2.98	Smooth and without burrs	Stabilization
Comparative example 5	8.23	3.02	Smooth and without burrs	Stabilization

As can be seen from table 4, the antibacterial agent inks of example 1 and comparative example 5 both had good stability, were smooth in printing process, and did not generate burrs. Compared with the comparative example 5, the pH value of the liquid medicine prepared in the example 1 is closer to neutral, and the liquid medicine is less corrosive to an ink cavity and a spray head of a printer; the viscosity is lower, the nozzle is not easy to block, the loss to the printer is less, and the performance is better.

In conclusion, compared with the standard ink, the antibacterial medicine ink provided by the invention has more proper pH, lower viscosity and better applicability.

Application example 1

The application example provides a method for preparing an ester drug sensitive test strip, which specifically comprises the following steps:

repeatedly cleaning an ink box of an ink-jet printer by deionized water for many times until residual ink in the ink box is completely and thoroughly cleaned, and then putting the cleaned ink box into a drying oven at 50 ℃ to dry to obtain a blank ink box;

the antibacterial ink prepared in example 1 and a blank ink (containing the humectant, the surfactant and the pH adjuster with the same concentration and species and containing no antibacterial) containing no antibacterial were respectively injected into an ink cartridge of an inkjet printer through a 0.22 μm filter membrane, and loaded into the inkjet printer;

printing the antibacterial drug ink and the blank ink on printing paper A4 of a preprinting substrate according to a preset proportion under the control of a computer, then drying for 3 minutes under the conditions of normal pressure, temperature of 28 ℃ and air humidity of 30%, slicing, packaging, sterilizing and storing to obtain the Etest drug sensitive test strip shown in figure 1. In the figure, the numbers in the top-down boxes indicate the percentage of the antimicrobial ink.

Application examples 2 to 6

The antibacterial ink prepared in example 1 used in application example 1 was replaced with the antibacterial ink prepared in examples 2 to 6.

Application example 7

In the application example, the matrix material of the Etest drug sensitive test strip is replaced by a transparent plastic sheet, and the other steps are consistent with the application example 1.

Application comparative examples 1 to 5

The antibacterial ink prepared in example 1 used in application example 1 was replaced with the antibacterial ink prepared in comparative examples 1 to 5.

Drug sensitive activity test

1) 42g of MH agar and 20g of glucose were weighed, dissolved in 1000mL of distilled water under heating, and dispensed into 2 Erlenmeyer flasks. Autoclaved at 121 ℃ for 15 minutes. Cooled to 50 ℃ and in a biosafety cabinet, the medium in the Erlenmeyer flask was poured into sterile plates, and 20mL of medium was poured into each plate and kept at 4 ℃ until use.

2) And (3) detection: in the biological safety cabinet, a sterile cotton stick is dipped with candida albicans bacterial liquid and coated on a plate for three times, so that the bacterial liquid is uniformly coated. The drug sensitive test strips of application examples 1 to 7 and application comparative examples 1 to 4 were attached to a dish with sterile forceps, and air bubbles were gently squeezed out.

3) Culturing at 35 deg.C for 24 h.

And (4) interpretation of results: the concentration of the drug at the junction of the inhibition zone and the drug sensitive paper sheet is the MIC value.

The test results are shown in table 5.

TABLE 5

	Printing effect	Accuracy of MIC values
			Application example 1	Good effect	Accurate and accurate
Application example 2	Print thumbnail displayMoisture content	Accurate and accurate
			Application example 3	Drying was rapid and the spray head occasionally clogged	Accurate and accurate
Application example 4	Easily block up shower nozzle	Has an error
			Application example 5	Not good enough	Has an error
Application example 6	Easily block up shower nozzle	Has an error
			Application example 7	Long drying time and difficult operation	Is more accurate
Application comparative example 1	Easily block up shower nozzle	Has an error
			Comparative application example 2	Not good enough	Has an error
Comparative application example 3	Easily block up shower nozzle	Has an error
			Application comparative example 4	Moist and not easy to dry	Has an error
Comparative application example 5	Good effect	-

As can be seen from Table 5, the test strips prepared in application examples 1-3 have good performance and accurate detection results; the test strip in the application example 1 also has a good printing effect, the printed part in the application example 2 is slightly moist, the test strip in the application example 3 has a good printing effect, but occasionally causes the blockage of the nozzle, and the application example 1 is more appropriate in formula, better in printing effect and less in damage to printing equipment compared with the application examples 2-3;

compared with application example 1 and application comparative examples 1-4, the humectant and the surfactant in the antibacterial drug ink disclosed by the invention are all absent, and the humectant and the surfactant are matched, so that the ink is easy to wet the surface of paper, the antibacterial drug is easy to combine on a drug sensitive test strip, the printing effect on the paper is improved, meanwhile, the drying speed of the ink can be adjusted, the ink is not broken or lack of strokes in the printing process, and the probability of blocking a spray head is reduced; if the humectant or the surfactant is not added, the detection result is possibly inaccurate;

as can be seen from the comparison between application example 1 and application example 4, the pH regulator affects the performance of the drug sensitive test strip, provides a proper pH value, is more beneficial to protecting the ink cavity and the nozzle of the printer, and can also increase the accuracy of the detection result;

as can be seen from comparison between application example 1 and application examples 5-6, sodium dodecyl benzene sulfonate and 1, 3-butanediol are preferred, and are matched, the obtained drug sensitive test strip has the optimal effect, and the loss to printing equipment is small;

as can be seen from comparison between application examples 1 and 7, the plastic sheet is used as a matrix material, so that the drying time of the test strip can be obviously prolonged, the test strip is inconvenient to take and use during drug sensitive detection, and bubbles are easily generated on the contact surface of a culture medium, so that the detection result is influenced to a certain extent; the printing paper has low production cost and convenient taking, can avoid bubbles generated when the culture medium is contacted with the test strip, has better diffusion effect of the antibacterial drug in the drug sensitive test strip in the culture medium, has more accurate test result and is more convenient to use;

the original ink of the printer is applied to the comparative example 5, has good printing effect, is only used for comparing the performances of the ink, and is not used for detecting the MIC value.

In conclusion, the best printing effect can be produced only by simultaneously adding the humectant, the surfactant and the pH regulator, selecting a proper reagent type and controlling the adding amount within a reasonable range; the absence of any one of the humectant, the surfactant, and the pH adjuster, or the use of an inappropriate reagent, or the actual amount added out of an appropriate range, has an effect on both the printing effect and the accuracy of the detection result.

In conclusion, the antibacterial drug ink and the method for preparing the Etest drug sensitive test strip based on the ink-jet printing method have important application value for preparing the Etest drug sensitive test strip, can obviously reduce the preparation cost and expand the application range of the Etest drug sensitive test strip.

The applicant declares that the above description is only a specific embodiment of the present invention, but the scope of the present invention is not limited thereto, and it should be understood by those skilled in the art that any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are within the scope and disclosure of the present invention.

Claims

1. An antibacterial drug ink for preparing an ester drug sensitive test strip, which is characterized by comprising the following components in percentage by mass: 0.01 to 10 percent of antibacterial drug, 0.5 to 5 percent of humectant, 0.5 to 2 percent of surfactant and the balance of deionized water.

2. The antibiotic drug ink according to claim 1, further comprising a pH adjusting agent of 0.1 to 0.2M;

preferably, the pH adjusting agent comprises any one or a combination of at least two of PBS, HEPES-NaOH or Tris-HCl, preferably Tris-HCl;

preferably, the pH value of the antibacterial medicine ink is 7.0-8.0.

3. The antimicrobial ink of claim 1 or 2, wherein the humectant comprises 1, 3-butanediol and/or 1, 2-propanediol, preferably 1, 3-butanediol;

preferably, the mass fraction of the humectant is 2-5%, preferably 3%;

preferably, the surfactant comprises sodium dodecylbenzene sulfonate;

preferably, the mass fraction of the surfactant is 1% to 2%, preferably 1.5%.

4. The antibacterial drug ink according to any one of claims 1 to 3, wherein the pH value of the antibacterial drug ink is 7.0 to 8.0, and the antibacterial drug ink comprises 0.01 to 10 mass percent of antibacterial drug, 2 to 5 mass percent of 1, 3-butanediol, 1 to 2 mass percent of sodium dodecyl benzene sulfonate, 0.1 to 0.2M molar content of Tris-HCl and deionized water.

5. Use of an antibacterial drug ink as claimed in any one of claims 1 to 4 in the preparation of an etet drug sensitive strip.

6. A method for preparing an ester drug sensitive test strip based on an ink-jet printing method, which is characterized by comprising the following steps:

(1) preparing the antibacterial drug ink as claimed in any one of claims 1 to 4, and injecting the antibacterial drug ink into an ink cartridge of an ink jet printer;

7. The method according to claim 6, wherein the step (1) of preparing the antibacterial ink further comprises the step of filtering the antibacterial ink;

8. The method according to claim 6 or 7, wherein the substrate material of step (2) comprises any one or a combination of at least two of printing paper, qualitative filter paper, fibrous film, polyacrylamide, polyester or polyamide, preferably printing paper;

preferably, the thickness of the printing paper is 0.03-0.2 mm, preferably 0.1 mm;

preferably, the drying in step (2) is specifically performed by: and (3) placing the substrate material containing the antibacterial drug ink in an environment with the air humidity less than 50%, and drying for 1-5 min at the temperature of 25-35 ℃.

9. The method according to any one of claims 6 to 8, wherein the sterilization in step (2) is irradiation sterilization;

preferably, the drying in the step (2) further comprises the operations of cutting and packaging;

10. A method according to any one of claims 6 to 9, characterized in that the method comprises the steps of:

(1) preparing the antibiotic drug ink as claimed in any one of claims 1 to 4, the antibiotic drug ink comprising: 0.01-10% of antibacterial agent, 2-5% of 1, 3-butanediol, 1-2% of sodium dodecyl benzene sulfonate, 0.1-0.2M of Tris-HCl and deionized water;