Method for evaluating sterilization effect of long-acting disinfectant

Abstract

The invention provides a sterilization effect evaluation method of a long-acting disinfectant, which comprises the following steps: taking a target strain, preparing a test suspension, and inoculating the test suspension on the surface of a target object to obtain a primary polluted object; taking a disinfectant to be detected, and applying the disinfectant to the primary contaminated object to obtain a target disinfected object; wearing the target disinfection object, and inoculating the test suspension to obtain a secondary contaminated object; and counting the obtained target strains of the secondary pollutants, and evaluating the sterilization effect of the to-be-detected disinfectant according to a counting result. According to the sterilization effect evaluation method of the long-acting disinfectant, the secondary pollution and abrasion states of the article are simulated, and the sterilization effect of the long-acting disinfectant on the article in actual use can be evaluated.

Description

Technical Field

[0001] The present invention relates to the technical field of disinfectant detection, and in particular to a method for evaluating the bactericidal effect of a long-acting disinfectant. Background Art

[0002] Long-acting disinfectants have gradually become one of the important directions for exploring innovative disinfectants worldwide because of their advantages such as reducing the frequency of disinfection, maintaining the sterilization rate continuously to improve the ability to prevent infection, reducing the difference between the effects of disinfection operations, increasing the utilization rate of drugs, and reducing manual workload. More and more disinfectants need to be evaluated for their long-term bactericidal effects.

[0003] At present, the method for evaluating the effect of long-acting disinfectants is mainly to apply it to the surface of an object, leave it for a certain period of time, and then test the bactericidal effect. When this method is used for evaluation, the surface of the object does not change after the disinfectant is applied, and it is a static process. However, during the normal use of the object, its surface is likely to continue to be worn or even removed by touching, wiping, etc., and there is also the possibility that the surface of the disinfected object will be contaminated again. Therefore, the above static effect evaluation method is difficult to evaluate the effect of long-acting disinfectants under actual conditions. Summary of the invention

[0004] The purpose of the present invention is to provide a method for evaluating the bactericidal effect of a long-acting disinfectant, so as to solve the technical problem that the effect evaluation method of the long-acting disinfectant in the prior art is difficult to obtain the bactericidal effect of the long-acting disinfectant on the objects in actual use.

[0005] In order to solve the above technical problems, the technical solution of the present invention is as follows:

[0006] A method for evaluating the bactericidal effect of a long-acting disinfectant comprises the following steps:

[0007] (1) taking a target bacterial species, preparing a test suspension, and then inoculating the test suspension onto the surface of a target object to obtain a primary contaminated object;

[0008] (2) taking a disinfectant to be tested and applying it to the primary contaminated object in step (1) to obtain a target disinfection object;

[0009] (3) abrading the target disinfection object obtained in step (2), inoculating the test suspension, drying, abrading, inoculating the test suspension again, drying, and repeating the cycle 3-5 times to obtain a secondary contaminated object;

[0010] (4) Counting the target bacteria species of the secondary contamination obtained in step (3), and evaluating the bactericidal effect of the disinfectant to be tested based on the counting results.

[0011] Preferably, in step (1), the test suspension is obtained by the following method:

[0012] Take the primary culture of the target bacteria, inoculate it in the culture medium, and perform subculture; use a diluent to wash the surface of the subculture culture medium, and shake the obtained suspension; take the suspension after shaking, and use a diluent to adjust the number of cells in the suspension to 1.5×10 8 cfu / ml-5.0×10 8 cfu / ml, and the test suspension was obtained;

[0013] Preferably, the culture medium is Tryptone Soy Agar (TSA).

[0014] Optionally, the test suspension comprises a bacterial test suspension or a yeast test suspension.

[0015] Preferably, the solvent of the diluent is water, and every 1000 ml of water contains 1.0-1.5 g of trypsin and 8.0-9.0 g of sodium chloride.

[0016] Preferably, interfering substances are also added to the test suspension.

[0017] Preferably, the interfering substance comprises bovine serum albumin; the concentration of the bovine serum albumin in the test suspension is 2.5-3.5 g / l.

[0018] Preferably, in step (1), the target object is one of a non-porous metal plate, a non-porous metal disk, a glass plate, and a PVC plate.

[0019] Preferably, in step (2), the disinfectant to be tested is one or more of polyhexamethylene biguanide hydrochloride and benzethonium chloride.

[0020] Preferably, the disinfectant to be tested is polyhexamethylene biguanide hydrochloride with a mass concentration of 0.65-1.2%.

[0021] Preferably, in step (3), abrading the target sterilization object includes dry abrasion and wet abrasion;

[0022] The dry abrasion comprises: taking a polypropylene wiping cloth, wiping the target disinfection object with a force of 2.0-2.5N;

[0023] The wet abrasion comprises: taking a polypropylene wiping cloth, wetting it with sterile water, and wiping the target disinfection object with a force of 2.0-2.5N.

[0024] Preferably, in step (3), the dry abrasion and wet abrasion are both rotation and grinding, specifically comprising: fixing the polypropylene wiping cloth on a counterweight block, and driving the counterweight block to rotate by a motor, so that the polypropylene wiping cloth rotates and grinds on the upper surface of the target disinfection object.

[0025] Preferably, the method for evaluating the bactericidal effect of a long-acting disinfectant further comprises a step of drying after the inoculation.

[0026] Preferably, the inoculation is carried out by spraying the test suspension onto the surface of the object.

[0027] Preferably, in step (4), counting the target bacteria species that are secondary contaminated obtained in step (3) specifically includes:

[0028] After the application time of the disinfectant to be tested reaches 168 hours, immersing the secondary contaminated material in a neutralizer and continuously immersing it at 20-25° C. for 5 min-10 min to obtain a mixed solution;

[0029] The mixed solution is diluted in a gradient manner and placed in sterile plates respectively, and cultured at 30-38° C. for 20-24 hours, and the number of colonies in the sterile plates is counted to obtain a counting result;

[0030] Wherein, the neutralizing agent includes one or more of glycine, polysorbate, saponin, lecithin, and sodium thiosulfate.

[0031] Preferably, the neutralizing agent comprises the following raw materials in parts by weight:

[0032] 100 parts water;

[0033] Glycine 0.5-1.0 part;

[0034] Polysorbate 2-3 parts;

[0035] Saponin 0.1-0.5 parts;

[0036] Lecithin 0.5-1.0 part;

[0037] Sodium thiosulfate 0.2-0.8 parts.

[0038] Preferably, the neutralizing agent comprises the following raw materials in parts by weight:

[0039] 100 parts water;

[0040] Glycine 0.8 part;

[0041] 2.8 parts of polysorbate;

[0042] Saponin 0.4 part;

[0043] 0.5 parts of lecithin;

[0044] 0.8 part of sodium thiosulfate;

[0045] 0.3 parts of fatty alcohol polyoxyethylene ether.

[0046] Preferably, step (4) further includes evaluating the bactericidal effect of the disinfectant to be tested in combination with the counting results of the blank control group.

[0047] The above solution of the present invention includes at least the following beneficial effects:

[0048] (1) The method for evaluating the bactericidal effect of a long-acting disinfectant of the present invention comprises the following steps: taking a target bacterial species, preparing a test suspension, and then inoculating the test suspension on the surface of a target object to obtain a primary contaminated object; taking a disinfectant to be tested, applying it to the primary contaminated object to obtain a target disinfected object; abrading the target disinfected object, and inoculating the test suspension to obtain a secondary contaminated object; counting the target bacterial species of the obtained secondary contaminated object, and evaluating the bactericidal effect of the disinfectant to be tested based on the counting result. The method for evaluating the bactericidal effect of a long-acting disinfectant of the present invention simulates the state of secondary contamination and abrasion, and can evaluate the bactericidal effect of a long-acting disinfectant on items in actual use.

[0049] (2) The method for evaluating the bactericidal effect of the long-acting disinfectant of the present invention includes abrasion of the target disinfection object by dry abrasion and wet abrasion. The dry abrasion step design realizes the test of the wiping resistance of the long-acting disinfectant, and the dry abrasion step design realizes the test of the washability of the long-acting disinfectant.

[0050] (3) In the method for evaluating the bactericidal effect of a long-acting disinfectant of the present invention, the neutralizing agent used is one or more of glycine, polysorbate, saponin, lecithin, and sodium thiosulfate. The neutralizing effect is good for disinfectants containing polyhexamethylene biguanide hydrochloride and benzethonium chloride, and the influence of the residual of such disinfectants on the test accuracy of the bactericidal effect evaluation method can be effectively avoided. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] Figure 1 It is an exploded diagram of the equipment structure of the method for evaluating the bactericidal effect of the long-acting disinfectant of the present invention;

[0052] Among them, 1-motor; 2-counterweight; 3-polypropylene wiping cloth; 4-target disinfection object. DETAILED DESCRIPTION

[0053] The exemplary embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. Although the exemplary embodiments of the present invention are shown in the accompanying drawings, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided in order to enable a more thorough understanding of the present invention and to enable the scope of the present invention to be fully communicated to those skilled in the art.

[0054] If the specific conditions are not specified in the embodiments of the present invention, the conventional conditions or the conditions recommended by the manufacturer are used. If the manufacturer of the reagents or instruments is not specified, they are all conventional products that can be obtained commercially. Only some of the reagents used in the embodiments of the present invention are provided below. Raw materials of different manufacturers and models do not affect the implementation of the technical solution of the present invention and the realization of the technical effects.

[0055] Example 1

[0056] The method for evaluating the bactericidal effect of the long-acting disinfectant of this embodiment comprises the following steps:

[0057] (1) taking a target bacterial species, preparing a test suspension, and then inoculating the test suspension onto the surface of a target object, drying it, and obtaining a primary contaminated object;

[0058] The test suspension is obtained by the following method: taking the primary culture of the target bacteria, inoculating it in a tryptone soy agar medium (TSA), and performing subculture; using a diluent to wash the surface of the subculture medium, and shaking the obtained suspension; taking the suspension after shaking, and adjusting the number of cells in the suspension to 1.5×10 8 cfu / ml, and the test suspension was obtained;

[0059] It should be noted that the test suspension can be a bacterial test suspension or a yeast test suspension, and those skilled in the art can select a suitable bacterial species according to the detection needs; in this embodiment, the target bacterial species is Escherichia coli.

[0060] The solvent of the diluent is water, and every 1000 ml of water contains 1.2 g of trypsin, 8.0 g of sodium chloride, and 3.0 g of interfering substance bovine serum albumin (BSA). The target object is a non-porous metal plate, which is circular in shape, 24 mm in diameter, and 1 mm in thickness. The inoculation is to inoculate 1 ml of the test suspension on the surface of the object by spraying (the same below, which will not be repeated in the following text).

[0061] (2) taking a disinfectant to be tested, applying it to the primary contaminated material in step (1) for 15 minutes to obtain a target disinfected object; the disinfectant to be tested is a polyhexamethylene biguanide hydrochloride solution with a mass concentration of 0.65%.

[0062] (3) Dry-abrading the target disinfection object obtained in step (2) and inoculating the test suspension. After drying, wet-abrading the target disinfection object and inoculating the test suspension again. After drying, the cycle is repeated three times to obtain a secondary contaminated object.

[0063] The dry abrasion includes: taking a polypropylene wiping cloth, wiping the target disinfection object with a force of 2.2N; the wet abrasion includes: taking a polypropylene wiping cloth, wetting it with sterile water, and wiping the target disinfection object with a force of 2.2N.

[0064] As a preferred implementation of this embodiment, Figure 1 As shown in FIG. 1 , it is an exploded view of the device structure of the method for evaluating the bactericidal effect of a long-acting disinfectant of the present invention, and the structure includes a motor 1, a counterweight block 2 connected to the output shaft of the motor 1, a polypropylene wiping cloth 3 fixed to the lower surface of the counterweight block 2, and a target disinfection object located below the polypropylene wiping cloth 3. The motor 1 provides a rotational force and applies a force in the direction of the arrow in the figure, so that the polypropylene wiping cloth 3 wipes the upper surface of the target disinfection object 4 with a force of 2.2N.

[0065] Wiping the target disinfection object with a force of 2.2N is achieved by rotating and grinding the above-mentioned structure, specifically including: fixing the polypropylene wiping cloth 3 on the counterweight block 2, and driving the counterweight block 2 to rotate by the motor 1, so that the polypropylene wiping cloth 3 rotates and grinds on the upper surface of the target disinfection object 4, thereby achieving wiping with a force of 2.2N.

[0066] (4) After the application time of the disinfectant to be tested reaches 168 hours, immerse the secondary contaminant obtained in step (3) in a neutralizer and continue to immerse at 20°C for 8 minutes to obtain a mixed solution; the mixed solution is gradient diluted and placed in sterile plates respectively, and cultured at 30°C for 22 hours, and the number of colonies in the sterile plates is counted to obtain a counting result; based on the counting result and in combination with the counting result of the blank control group, the bactericidal effect of the disinfectant to be tested is evaluated.

[0067] Wherein, the neutralizing agent comprises the following raw materials in parts by weight:

[0068] 100 parts water;

[0069] Glycine 0.8 part;

[0070] 2 parts of polysorbate;

[0071] Saponin 0.5 part;

[0072] 0.5 parts of lecithin;

[0073] 0.5 parts of sodium thiosulfate.

[0074] The counting result of the blank control group was obtained by the following method:

[0075] (1') taking a target bacterial species, preparing a test suspension, and then inoculating the test suspension onto the surface of a target object, drying, and obtaining a primary contaminated object;

[0076] (2') taking sterile water, and applying the sterile water to the primary contaminated object in step (1') to obtain a blank group of sterilized objects;

[0077] (3') dry-abrading the sterilized blank object obtained in step (2'), inoculating the test suspension, and then wet-abrading the blank object after drying, and then inoculating the test suspension. After drying, the cycle is repeated three times to obtain a secondary contaminated object.

[0078] (4') After the sterile water is applied for 168 hours, the secondary contaminant obtained in step (3') is immersed in a neutralizer and continuously immersed at 20°C for 8 minutes to obtain a mixed solution; the mixed solution is gradiently diluted and placed in sterile plates respectively, and cultured at 30°C for 22 hours, and the number of colonies in the sterile plates is counted to obtain the counting result of the blank control group.

[0079] The counting result of the blank control group is obtained in the same manner as the counting result obtained in step (4) of this embodiment, with the only difference being that in step (2), the disinfectant to be tested is replaced with sterile water.

[0080] Example 2

[0081] The method for evaluating the bactericidal effect of the long-acting disinfectant of this embodiment comprises the following steps:

[0082] (1) taking a target bacterial species, preparing a test suspension, and then inoculating the test suspension onto the surface of a target object, drying it, and obtaining a primary contaminated object;

[0083] The test suspension is obtained by the following method: taking the primary culture of the target bacterial species, inoculating it in the culture medium, and performing subculture; using a diluent to wash the surface of the subculture culture medium, and shaking the obtained suspension; taking the suspension after shaking, and adjusting the number of cells in the suspension to 5.0×10 8 cfu / ml, and the test suspension was obtained;

[0084] It should be noted that the test suspension can be a bacterial test suspension or a yeast test suspension, and those skilled in the art can select a suitable bacterial species according to the detection needs; in this embodiment, the target bacterial species is Escherichia coli.

[0085] The solvent of the diluent is water, and every 1000 ml of water contains 1.5 g of trypsin, 8.5 g of sodium chloride, and 3.5 g of interfering substance bovine serum albumin (BSA). The target object is a non-porous metal disk with a diameter of 24 mm and a thickness of 1 mm. The inoculation is to inoculate 1 ml of the test suspension on the surface of the object by spraying (the same below, which will not be repeated in the following text).

[0086] (2) taking a disinfectant to be tested, applying it to the primary contaminated material in step (1) for 5 minutes, and obtaining a target disinfected object; the disinfectant to be tested is a benzethonium chloride solution with a mass concentration of 0.3%.

[0087] (3) Dry-abrading the target disinfection object obtained in step (2) and inoculating the test suspension. After drying, wet-abrading the target disinfection object and inoculating the test suspension again. After drying, the cycle is repeated 5 times to obtain a secondary contaminated object.

[0088] The dry abrasion includes: taking a polypropylene wiping cloth, wiping the target disinfection object with a force of 2.5N; the wet abrasion includes: taking a polypropylene wiping cloth, wetting it with sterile water, and wiping the target disinfection object with a force of 2.5N.

[0089] The target disinfection object is wiped with a force of 2.5N. Specifically, the rotation and grinding are performed using the same equipment structure as in Example 1. Figure 1 .

[0090] (4) After the application time of the disinfectant to be tested reaches 168 hours, immerse the secondary contaminated material obtained in step (3) in a neutralizer and continue to immerse it at 25°C for 10 minutes to obtain a mixed solution; the mixed solution is gradient diluted and placed in sterile plates respectively, and cultured at 38°C for 24 hours, and the number of colonies in the sterile plates is counted to obtain a counting result; based on the counting result and in combination with the counting result of the blank control group, the bactericidal effect of the disinfectant to be tested is evaluated.

[0091] Wherein, the neutralizing agent comprises the following raw materials in parts by weight:

[0092] 100 parts water;

[0093] 1.0 part of glycine;

[0094] Polysorbate 3 parts;

[0095] Saponin 0.3 part;

[0096] 1.0 part of lecithin;

[0097] 0.2 parts of sodium thiosulfate.

[0098] The counting result of the blank control group is obtained in the same manner as the counting result obtained in step (4) of this embodiment, with the only difference being that in step (2), the disinfectant to be tested is replaced with sterile water.

[0099] Example 3

[0100] The method for evaluating the bactericidal effect of the long-acting disinfectant of this embodiment comprises the following steps:

[0101] (1) taking a target bacterial species, preparing a test suspension, and then inoculating the test suspension onto the surface of a target object, drying it, and obtaining a primary contaminated object;

[0102] The test suspension is obtained by the following method: taking the primary culture of the target bacteria, inoculating it in the culture medium, and performing subculture; using a diluent to wash the surface of the subculture culture medium, and shaking the obtained suspension; taking the suspension after shaking, and adjusting the number of cells in the suspension to 3.2×10 8 cfu / ml, and the test suspension was obtained;

[0103] It should be noted that the test suspension can be a bacterial test suspension or a yeast test suspension, and those skilled in the art can select a suitable bacterial species according to the detection needs; in this embodiment, the target bacterial species is Escherichia coli.

[0104] The solvent of the diluent is water, and every 1000 ml of water contains 1.0 g of trypsin, 9.0 g of sodium chloride, and 2.5 g of interfering substance bovine serum albumin (BSA). The target object is a glass plate, which is circular in shape, 24 mm in diameter, and 1 mm in thickness. The inoculation is to inoculate 1 ml of the test suspension on the surface of the object by spraying (the same below, which will not be repeated in the following text).

[0105] (2) taking a disinfectant to be tested, applying it to the primary contaminated material in step (1) for 5 minutes, and obtaining a target disinfected object; the disinfectant to be tested is a mixture of a polyhexamethylene biguanide hydrochloride solution and a benzethonium chloride solution. The mass concentration of the polyhexamethylene biguanide hydrochloride is 1.2%; the mass concentration of the benzethonium chloride is 0.3%.

[0106] (3) Dry-abrading the target disinfection object obtained in step (2) and inoculating the test suspension. After drying, wet-abrading the target disinfection object and inoculating the test suspension again are performed. After drying, the cycle is repeated 4 times to obtain a secondary contaminated object.

[0107] The dry abrasion includes: taking a polypropylene wiping cloth, wiping the target disinfection object with a force of 2.0N; the wet abrasion includes: taking a polypropylene wiping cloth, wetting it with sterile water, and wiping the target disinfection object with a force of 2.0N.

[0108] The target disinfection object is wiped with a force of 2.0 N. Specifically, the rotation and grinding are performed using the same equipment structure as in Example 1. Figure 1 .

[0109] (4) After the application time of the disinfectant to be tested reaches 168 hours, immerse the secondary contaminant obtained in step (3) in a neutralizer and continue to immerse at 22°C for 5 minutes to obtain a mixed solution; the mixed solution is gradient diluted and placed in sterile plates respectively, and cultured at 34°C for 20 hours, and the number of colonies in the sterile plates is counted to obtain a counting result; based on the counting result and in combination with the counting result of the blank control group, the bactericidal effect of the disinfectant to be tested is evaluated.

[0110] Wherein, the neutralizing agent comprises the following raw materials in parts by weight:

[0111] 100 parts water;

[0112] 0.5 part of glycine;

[0113] 2.5 parts of polysorbate;

[0114] Saponin 0.1 part;

[0115] 0.8 parts of lecithin;

[0116] 0.8 parts of sodium thiosulfate.

[0117] The counting result of the blank control group is obtained in the same manner as the counting result obtained in step (4) of this embodiment, with the only difference being that in step (2), the disinfectant to be tested is replaced with sterile water.

[0118] Example 4

[0119] The method for evaluating the bactericidal effect of the long-acting disinfectant of this embodiment comprises the following steps:

[0120] (1) taking a target bacterial species, preparing a test suspension, and then inoculating the test suspension onto the surface of a target object, drying it, and obtaining a primary contaminated object;

[0121] The test suspension is obtained by the following method: taking the primary culture of the target bacteria, inoculating it in the culture medium, and performing subculture; using a diluent to wash the surface of the subculture culture medium, and shaking the obtained suspension; taking the suspension after shaking, and adjusting the number of cells in the suspension to 3.0×10 8 cfu / ml, and the test suspension was obtained;

[0122] It should be noted that the test suspension can be a bacterial test suspension or a yeast test suspension, and those skilled in the art can select a suitable bacterial species according to the detection needs; in this embodiment, the target bacterial species is Escherichia coli.

[0123] The solvent of the diluent is water, and every 1000 ml of water contains 1.2 g of trypsin, 8.5 g of sodium chloride, and 3.0 g of interfering substance bovine serum albumin (BSA). The target object is a PVC plate, which is circular in shape, 24 mm in diameter, and 1 mm in thickness. The inoculation is to inoculate 1 ml of the test suspension on the surface of the object by spraying (the same below, which will not be repeated in the following text).

[0124] (2) taking a disinfectant to be tested, applying it to the primary contaminated material in step (1) for 5 minutes, and obtaining a target disinfected object; the disinfectant to be tested is a mixture of a polyhexamethylene biguanide hydrochloride solution and a benzethonium chloride solution. The mass concentration of the polyhexamethylene biguanide hydrochloride is 1.2%; the mass concentration of the benzethonium chloride is 0.3%.

[0125] (3) Dry-abrading the target disinfection object obtained in step (2) and inoculating the test suspension. After drying, wet-abrading the target disinfection object and inoculating the test suspension again. After drying, the cycle is repeated 5 times to obtain a secondary contaminated object.

[0126] The dry abrasion includes: taking a polypropylene wiping cloth, wiping the target disinfection object with a force of 2.5N; the wet abrasion includes: taking a polypropylene wiping cloth, wetting it with sterile water, and wiping the target disinfection object with a force of 2.5N.

[0127] The target disinfection object is wiped with a force of 2.5N. Specifically, the rotation and grinding are performed using the same equipment structure as in Example 1. Figure 1 .

[0128] It should be noted that the cycle repetition means that the target disinfection object is dry-abraded and inoculated with the test suspension, and after drying, it is wet-abraded and then inoculated with the test suspension, and after drying, it is one cycle.

[0129] (4) After the application time of the disinfectant to be tested reaches 168 hours, immerse the secondary contaminated material obtained in step (3) in a neutralizer and soak it for 10 minutes at 25°C to obtain a mixed solution; the mixed solution is diluted in a gradient manner and placed in sterile plates respectively, and cultured at 37°C for 24 hours, and the number of colonies in the sterile plates is counted to obtain a counting result; based on the counting result and in combination with the counting result of the blank control group, the bactericidal effect of the disinfectant to be tested is evaluated.

[0130] Wherein, the neutralizing agent comprises the following raw materials in parts by weight:

[0131] 100 parts water;

[0132] Glycine 0.8 part;

[0133] Polysorbate 2.8 parts;

[0134] Saponin 0.4 part;

[0135] 0.5 parts of lecithin;

[0136] 0.8 parts of sodium thiosulfate.

[0137] The counting result of the blank control group is obtained in the same manner as the counting result obtained in step (4) of this embodiment, with the only difference being that in step (2), the disinfectant to be tested is replaced with sterile water.

[0138] Example 5

[0139] The method for evaluating the bactericidal effect of the long-acting disinfectant of this embodiment is the same as that of Embodiment 4, except that the neutralizing agent used in step (4) comprises the following raw materials in parts by weight:

[0140] 100 parts water;

[0141] Glycine 0.8 part;

[0142] Polysorbate 2.8 parts;

[0143] Saponin 0.4 part;

[0144] 0.5 parts of lecithin.

[0145] Example 6

[0146] The method for evaluating the bactericidal effect of the long-acting disinfectant of this embodiment is the same as that of Embodiment 4, except that the neutralizing agent used in step (4) comprises the following raw materials in parts by weight:

[0147] 100 parts water;

[0148] Glycine 0.8 part;

[0149] Polysorbate 2.8 parts;

[0150] Saponin 0.4 part;

[0151] 0.8 parts of sodium thiosulfate.

[0152] Example 7

[0153] The method for evaluating the bactericidal effect of the long-acting disinfectant of this embodiment is the same as that of Embodiment 4, except that the neutralizing agent used in step (4) comprises the following raw materials in parts by weight:

[0154] 100 parts water;

[0155] Glycine 0.8 part;

[0156] Polysorbate 2.8 parts;

[0157] 0.5 parts of lecithin;

[0158] 0.8 parts of sodium thiosulfate.

[0159] Example 8

[0160] The method for evaluating the bactericidal effect of the long-acting disinfectant of this embodiment is the same as that of Embodiment 4, except that the neutralizing agent used in step (4) comprises the following raw materials in parts by weight:

[0161] 100 parts water;

[0162] Glycine 0.8 part;

[0163] Saponin 0.4 part;

[0164] 0.5 parts of lecithin;

[0165] 0.8 parts of sodium thiosulfate.

[0166] Example 9

[0167] The method for evaluating the bactericidal effect of the long-acting disinfectant of this embodiment is the same as that of Embodiment 4, except that the neutralizing agent used in step (4) comprises the following raw materials in parts by weight:

[0168] 100 parts water;

[0169] Polysorbate 2.8 parts;

[0170] Saponin 0.4 part;

[0171] 0.5 parts of lecithin;

[0172] 0.8 parts of sodium thiosulfate.

[0173] Example 10

[0174] The method for evaluating the bactericidal effect of the long-acting disinfectant of this embodiment is the same as that of Embodiment 4, except that the neutralizing agent used in step (4) comprises the following raw materials in parts by weight:

[0175] 100 parts water;

[0176] Glycine 0.8 part;

[0177] 2.8 parts of polysorbate;

[0178] Saponin 0.4 part;

[0179] 0.5 parts of lecithin;

[0180] 0.8 part of sodium thiosulfate;

[0181] 0.3 parts of fatty alcohol polyoxyethylene ether; wherein the fatty alcohol polyoxyethylene ether is referred to as emulsifier AEO-3, which is a condensation product of fatty alcohol and ethylene oxide.

[0182] Embodiment 11

[0183] The method for evaluating the bactericidal effect of the long-acting disinfectant of this embodiment is the same as that of Embodiment 4, except that the neutralizing agent used in step (4) comprises the following raw materials in parts by weight:

[0184] 100 parts water;

[0185] Glycine 0.8 part;

[0186] 2.8 parts of polysorbate;

[0187] Saponin 0.4 part;

[0188] 0.5 parts of lecithin;

[0189] 0.8 parts of sodium thiosulfate;

[0190] 0.1 parts of fatty alcohol polyoxyethylene ether.

[0191] Example 12

[0192] The method for evaluating the bactericidal effect of the long-acting disinfectant of this embodiment is the same as that of Embodiment 4, except that the neutralizing agent used in step (4) comprises the following raw materials in parts by weight:

[0193] 100 parts water;

[0194] Glycine 0.8 part;

[0195] 2.8 parts of polysorbate;

[0196] Saponin 0.4 part;

[0197] 0.5 parts of lecithin;

[0198] 0.8 parts of sodium thiosulfate;

[0199] 0.5 parts of fatty alcohol polyoxyethylene ether.

[0200] Effect experiment example

[0201] In order to verify the technical effect of the bactericidal effect evaluation method of the long-acting disinfectant described in the present invention, the following test was performed:

[0202] Experiment 1:

[0203] The bactericidal effect evaluation method of the long-acting disinfectant of Examples 1-12 was used to obtain the counting results, and the killing logarithm of Escherichia coli was calculated. The results were repeated 5 times, and the average killing rate of Escherichia coli 8099 (4th to 6th generations) was calculated, and the standard deviation was calculated.

[0204] Among them, the killing log value = the average log value of the blank control group - the average log value of the test group.

[0205] After testing, the following results were obtained:

[0206]

[0207] Experiment 2:

[0208] The bactericidal effect evaluation method of the long-acting disinfectant of Examples 1-12 was used to obtain the counting results, calculate the killing logarithm of Escherichia coli, repeat 5 times, calculate the average killing rate of Staphylococcus aureus ATCC6538 (5th-7th generations), and calculate the standard deviation value;

[0209] Among them, the killing log value = the average log value of the blank control group - the average log value of the test group.

[0210] After testing, the following results were obtained:

[0211]

[0212] Experiment 3:

[0213] Take the neutralizers in the method for evaluating the bactericidal effect of the long-acting disinfectant of Examples 1-12, and conduct neutralizer identification tests respectively. Take Escherichia coli as the verification strain, and use the carrier sample smeared with Escherichia coli to conduct the neutralizer identification test. The test is repeated three times, and the inter-group colony error rate of each neutralizer is calculated. Among them, the test groups are:

[0214] The first group is a neutralizer + bacterial sheet, in order to observe whether the neutralizer has an inhibitory effect on the growth of Escherichia coli; that is, a PVC plate with a circular shape, a diameter of 24 mm, and a thickness of 1 mm is immersed in the neutralizer, and the immersion is continued for 10 minutes at 25°C, and then the PVC plate is used as the target object according to step (1) of Example 4 to obtain the primary contaminated material, which is the bacterial sheet formed by inoculating Escherichia coli on the PVC plate.

[0215] The second group is disinfectant + neutralizer + bacterial slice, that is, a PVC plate with a circular shape, a diameter of 24 mm and a thickness of 1 mm is taken, immersed in the neutralizer, and continuously immersed for 10 minutes at 25°C. According to steps (1) and (2) of Example 4, the PVC plate is used as the target object to obtain the target disinfection object.

[0216] The third group is sodium chloride tryptone dilution solution + bacterial slices, that is, a PVC plate with a circular shape, a diameter of 24 mm, and a thickness of 1 mm is taken, and the PVC plate is used as the target object according to step (1) of Example 4 to obtain the primary contaminated material.

[0217] The calculation formula of the error rate of colony counts among the first, second and third groups is as follows:

[0218] The error rate of colony count between groups = the sum of the absolute values ​​of (average number of colonies between three groups - average number of colonies in each group) × 100%

[0219] After testing, the neutralizers in Examples 1-12 can effectively neutralize the test bacteria, and the neutralizers have no adverse effects on the test bacteria, and the following results are obtained:

[0220] Group Error rate among three groups Example 1 4.57％ Example 2 5.32％ Example 3 4.98％ Example 4 3.79％ Example 5 11.4％ Example 6 13.5％ Example 7 10.6％ Example 8 14.27％ Example 9 9.56％ Example 10 1.96％ Embodiment 11 2.35％ Example 12 2.79％

[0221] According to the above experimental results, the bactericidal effect evaluation method of the long-acting disinfectant of the present invention has good repeatability of experimental results. In particular, in Example 10, the standard deviation of the bactericidal effect evaluation results for Escherichia coli and Staphylococcus aureus is the smallest, and the repeatability is the best. By adopting the neutralizer in Example 10, the bactericidal effect evaluation result is also more accurate. In addition, the neutralizer in Example 10 also has the lowest error rate among the three groups. According to the comparison of Example 4 and Example 10, the addition of fatty alcohol polyoxyethylene ether enables the obtained neutralizer to better avoid the influence on the accuracy of the bactericidal effect evaluation method. However, according to the comparison of Examples 10-12, the addition of fatty alcohol polyoxyethylene ether is too large or too small, which will cause the performance of the neutralizer to decline. According to the comparison of Examples 1-4 and 5-9, the omission of each component of the neutralizer will affect the performance of the neutralizer to varying degrees.

[0222] The above is 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 principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for evaluating the bactericidal effect of a long-acting disinfectant, characterized in that: The steps include: (1) taking a target bacterial species, preparing a test suspension, and then inoculating the test suspension onto the surface of a target object to obtain a primary contaminated object; (2) taking a disinfectant to be tested and applying it to the primary contaminated object in step (1) to obtain a target disinfection object; (3) abrading the target disinfection object obtained in step (2), inoculating the test suspension, drying, abrading, inoculating the test suspension again, drying, and repeating the cycle 3-5 times to obtain a secondary contaminated object; (4) Counting the target bacteria species of the secondary contamination obtained in step (3), and evaluating the bactericidal effect of the disinfectant to be tested based on the counting results.

2. The method for evaluating the bactericidal effect of a long-acting disinfectant according to claim 1, characterized in that: In step (1), the test suspension is obtained by the following method: Take the primary culture of the target bacteria, inoculate it in the culture medium, and perform subculture; use a diluent to wash the surface of the subculture culture medium, and shake the obtained suspension; take the suspension after shaking, and use a diluent to adjust the number of cells in the suspension to 1.5×10 8 cfu / ml-5.0×10 8 cfu / ml, and the test suspension was obtained.

3. The method for evaluating the bactericidal effect of a long-acting disinfectant according to claim 2, characterized in that: The solvent of the diluent is water, and every 1000 ml of water contains 1.0-1.5 g of trypsin and 8.0-9.0 g of sodium chloride.

4. The method for evaluating the bactericidal effect of a long-acting disinfectant according to claim 1, characterized in that: In step (1), the target object is one of a non-porous metal plate, a non-porous metal disk, a glass plate, and a PVC plate.

5. The method for evaluating the bactericidal effect of a long-acting disinfectant according to claim 1, characterized in that: In step (3), the abrasion of the target disinfection object includes dry abrasion and wet abrasion; The dry abrasion comprises: taking a polypropylene wiping cloth, wiping the target disinfection object with a force of 2.0-2.5N; The wet abrasion comprises: taking a polypropylene wiping cloth, wetting it with sterile water, and wiping the target disinfection object with a force of 2.0-2.5N.

6. The method for evaluating the bactericidal effect of a long-acting disinfectant according to claim 5, characterized in that: In step (3), the dry wear and wet wear are both rotation and grinding, specifically including: fixing the polypropylene wiping cloth on a counterweight block, and driving the counterweight block to rotate by a motor, so that the polypropylene wiping cloth rotates and grinds on the upper surface of the target disinfected object.

7. The method for evaluating the bactericidal effect of a long-acting disinfectant according to claim 1, characterized in that: The method further comprises a drying step after the inoculation.

8. The method for evaluating the bactericidal effect of a long-acting disinfectant according to claim 7, characterized in that: In step (4), counting the target bacteria species that are secondary contaminated obtained in step (3) specifically includes: After the application time of the disinfectant to be tested reaches 168 hours, immersing the secondary contaminated material in a neutralizer and continuously immersing it at 20-25° C. for 5 min-10 min to obtain a mixed solution; The mixed solution is diluted in a gradient manner and placed in sterile plates respectively, and cultured at 30-38° C. for 20-24 hours, and the number of colonies in the sterile plates is counted to obtain a counting result; Wherein, the neutralizing agent includes one or more of glycine, polysorbate, saponin, lecithin, and sodium thiosulfate.

9. The method for evaluating the bactericidal effect of a long-acting disinfectant according to claim 8, characterized in that: The neutralizing agent comprises the following raw materials in parts by weight: 100 parts water; Glycine 0.5-1.0 part; Polysorbate 2-3 parts; Saponin 0.1-0.5 parts; Lecithin 0.5-1.0 part; Sodium thiosulfate 0.2-0.8 parts.

10. The method for evaluating the bactericidal effect of a long-acting disinfectant according to claim 1, characterized in that: Step (4) also includes evaluating the bactericidal effect of the disinfectant to be tested in combination with the counting results of the blank control group.

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