阅读说明：本技术 紫外线消毒盒杀菌率验证方法 (Method for verifying sterilization rate of ultraviolet disinfection box ) 是由 胡桂云 卢涛 李席如 周德发 王全胜 于 2020-12-05 设计创作，主要内容包括：本发明公开了一种紫外线消毒盒杀菌率验证方法,其先将多种培养基在高温高压蒸汽炉内处理灭菌备用,并制备浓度为0.9％的氯化钠稀释液、洗脱液；再对物体表面进行灭菌为无菌状态；接下来进行菌悬液的制备,然后吸取制备好的菌悬液于已灭菌的物体表面最难杀菌的位置,进行菌片制备；再将染菌物体分别进行紫外线消毒盒试验组和阳性对照组的试验,获取菌数,最后根据杀菌率进行判定是否合格,通过如此杀菌率验证步骤进行,以达到准确高效的验证紫外线消毒盒杀菌率的有益效果。(The invention discloses a sterilization rate verification method of an ultraviolet sterilization box, which comprises the steps of treating and sterilizing various culture mediums in a high-temperature high-pressure steam furnace for later use, and preparing 0.9% sodium chloride diluent and eluent; then sterilizing the surface of the object to be in an aseptic state; preparing bacterial suspension, sucking the prepared bacterial suspension to a position on the surface of a sterilized object which is most difficult to sterilize, and preparing bacterial tablets; and then, respectively testing the infected objects with the ultraviolet disinfection box test group and the positive control group to obtain the number of bacteria, and finally judging whether the infected objects are qualified according to the sterilization rate, wherein the sterilization rate verification step is carried out so as to achieve the beneficial effect of accurately and efficiently verifying the sterilization rate of the ultraviolet disinfection box.)

1. A sterilization rate verification method for an ultraviolet disinfection box is characterized by comprising the following steps:

s1: preparing a sterilization culture medium and a reagent: treating and sterilizing a tryptone soytone liquid culture medium, a Sabouraud's dextrose liquid culture medium, a tryptone soytone agar culture medium and a Sabouraud's dextrose agar culture medium in a high-temperature high-pressure steam furnace for later use, and preparing a sodium chloride diluent and an eluent with the concentration of 0.9 percent;

s2: sterilizing the surface of the object; cleaning and drying an object to be sterilized, sterilizing the object at high temperature and high pressure, and detecting the surface of the object to be sterilized to be in an aseptic state;

s3: preparing a bacterial suspension: preparing a bacteria test tube inclined plane by a bacteria passage method, inoculating a fresh culture of scraped bacteria into a sodium chloride test tube filled with 5mL of bacteria suspension with the concentration, roughly measuring the bacteria concentration of the bacteria suspension with a Mach turbidimeter, counting the viable bacteria of the bacteria solution, diluting the bacteria suspension into the bacteria suspension with the bacteria content of 5 multiplied by 105 to 5 multiplied by 106cfu/mL by using sodium chloride diluent with the concentration, and determining the number of bacterial colonies;

s4: preparing a fungus tablet: sucking the bacterial suspension obtained in the dripping step S3 into the position where the surface of the object sterilized in the step S2 is the most difficult to sterilize, placing the object flat in a sterilization plate, and then placing the object in a 37 ℃ thermostat for 30min for drying treatment;

s5: bacterial count experiments: is divided into an ultraviolet disinfection box test group and a positive control group, wherein,

the ultraviolet disinfection box test group is as follows: placing the contaminated object in the step S4 into an ultraviolet disinfection box for disinfection, then eluting the sterilized object with a sodium chloride reagent with the concentration, sucking the eluted object into a culture dish, pouring the eluted object as a flat plate, inoculating two flat plates for culture, and counting the flat plates;

the positive control group was: directly placing the infected objects in the step S4 in a sodium chloride reagent with the concentration for elution, sucking the objects into a culture dish for pouring as a flat plate, inoculating two flat plates for culture, and counting the flat plates;

s6: and judging the result, according to a calculation formula of the sterilization rate:

and judging that the sterilization rate reaches the target, and determining that the sterilization rate is qualified.

2. The ultraviolet sterilization box sterilization rate verification method of claim 1, wherein the temperature of the high-temperature high-pressure steam oven in the step S1 is 121 ℃ and the time is 20 min.

3. The method for verifying the sterilization rate of an ultraviolet disinfection box as claimed in claim 1, wherein the bacterial suspension prepared by the bacterial suspension preparation method in step S3 comprises a bacterial suspension of staphylococcus aureus, a bacterial suspension of escherichia coli, a bacterial suspension of pseudomonas aeruginosa and a bacterial suspension of candida albicans.

4. The method for verifying the sterilization rate of an ultraviolet disinfection box according to claim 3, wherein 0.1mL of bacterial suspension with the bacterial content of 5 x 106cfu/mL is sucked on the surface of the sterilized object in step S4, the dripping area is 2 x 3cm, and the bacterial content of the recovered carrier is 5 x 105 cfu/piece.

5. The ultraviolet disinfection box sterilization rate verification method of claim 4, wherein in step S4, the surface of the sterilized object is inoculated with bacterial strains of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Candida albicans.

6. The ultraviolet disinfection box sterilization rate verification method of claim 5, wherein in step S5, the ultraviolet disinfection box test group puts the infected objects into the ultraviolet disinfection box for disinfection for 60S, and uses 100ml of 0.9% sodium chloride reagent for elution, and sucks 1ml into the culture dish.

7. The ultraviolet disinfection box sterilization rate verification method of claim 6, wherein in step S5, the ratio of the bacterial propagules: the culture temperature of the staphylococcus aureus, the escherichia coli and the pseudomonas aeruginosa is 30-37 ℃, and the culture time is 3 days; fungi: the culture temperature of the candida albicans is 20-25 ℃, and the culture time is 5 days.

Technical Field

The invention relates to the technical field of sanitation sterilization, in particular to a method for verifying the sterilization rate of an ultraviolet sterilization box.

Background

Along with the improvement of the consciousness of people for living health prevention, clothes, food, live and walk in life are required to meet the health standard after disinfection and sterilization, for example, people often carry out alcohol disinfection treatment on mobile phones and glasses, and carry out ultraviolet irradiation disinfection treatment on clothes and food, and the like, so as to prevent physical health through disinfection and sterilization measures.

At present, the ultraviolet sterilization lamp is used for irradiation for disinfection and sterilization, is one of the common domestic physical disinfection methods, and is widely applied to scientific research and production departments of medicine, health, biological products and the like for disinfection treatment of air, water and object surfaces. Referring to the iodine-containing disinfectant hygienic standard, the sterilization rate of bacterial propagules is more than 99.999 percent and the sterilization rate of fungi is more than 99.99 percent after ultraviolet sterilization is designed to reach the medical grade sterilization level, so that the verification of the ultraviolet sterilization rate is crucial, and the product sterilization rate is qualified only when the sterilization rate is qualified in the place where the bacteria are most difficult to kill.

Disclosure of Invention

In order to solve the problems, the invention provides a method for verifying the sterilization rate of an ultraviolet disinfection box, which can accurately and efficiently verify the sterilization rate of the ultraviolet disinfection box.

Based on the above, the invention provides a method for verifying the sterilization rate of an ultraviolet disinfection box, which comprises the following steps:

s1: preparing a sterilization culture medium and a reagent: treating and sterilizing a tryptone soytone liquid culture medium, a Sabouraud's dextrose liquid culture medium, a tryptone soytone agar culture medium and a Sabouraud's dextrose agar culture medium in a high-temperature high-pressure steam furnace for later use, and preparing a sodium chloride diluent and an eluent with the concentration of 0.9 percent;

s2: sterilizing the surface of the object; cleaning and drying an object to be sterilized, sterilizing the object at high temperature and high pressure, and detecting the surface of the object to be sterilized to be in an aseptic state;

s3: preparing a bacterial suspension: preparing a bacteria test tube inclined plane by a bacteria passage method, inoculating a fresh culture of scraped bacteria into a sodium chloride test tube filled with 5mL of bacteria suspension with the concentration, roughly measuring the bacteria concentration of the bacteria suspension with a Mach turbidimeter, counting the viable bacteria of the bacteria solution, diluting the bacteria suspension into the bacteria suspension with the bacteria content of 5 multiplied by 105 to 5 multiplied by 106cfu/mL by using sodium chloride diluent with the concentration, and determining the number of bacterial colonies;

s4: preparing a fungus tablet: sucking the bacterial suspension obtained in the dripping step S3 into the position where the surface of the object sterilized in the step S2 is the most difficult to sterilize, placing the object flat in a sterilization plate, and then placing the object in a 37 ℃ thermostat for 30min for drying treatment;

s5: bacterial count experiments: the kit is divided into an ultraviolet disinfection box test group and a positive control group, wherein the ultraviolet disinfection box test group comprises: placing the contaminated object in the step S4 into an ultraviolet disinfection box for disinfection, then eluting the sterilized object with a sodium chloride reagent with the concentration, sucking the eluted object into a culture dish, pouring the eluted object as a flat plate, inoculating two flat plates for culture, and counting the flat plates; the positive control group was: directly placing the infected objects in the step S4 in a sodium chloride reagent with the concentration for elution, sucking the objects into a culture dish for pouring as a flat plate, inoculating two flat plates for culture, and counting the flat plates;

s6: and judging the result, according to a calculation formula of the sterilization rate:

and judging that the sterilization rate reaches the target, and determining that the sterilization rate is qualified.

In the above technical solution, the temperature of the high-temperature high-pressure steam furnace in the step S1 is 121 ℃, and the time is 20 min.

In the above technical solution, the bacterial suspension prepared by the bacterial suspension preparation method in step S3 includes a bacterial suspension of staphylococcus aureus, a bacterial suspension of escherichia coli, a bacterial suspension of pseudomonas aeruginosa, and a bacterial suspension of candida albicans.

In the above technical solution, in the step S4, 0.1mL of bacterial suspension with a bacterial content of 5 × 106cfu/mL is sucked on the surface of the sterilized object, the dripping area is 2 × 3cm, and the bacterial content of the carrier recovery is 5 × 105 cfu/carrier recovery.

In the above technical solution, in the step S4, the surface of the sterilized object is inoculated with bacterial strains of staphylococcus aureus, escherichia coli, pseudomonas aeruginosa and candida albicans respectively.

In the above technical solution, in step S5, the ultraviolet disinfection box test group puts the contaminated object into the ultraviolet disinfection box for disinfection for 60S, and uses 100ml of 0.9% sodium chloride reagent for elution, and sucks 1ml of the contaminated object into the culture dish.

In the above technical solution, in the step S5, the ratio of the bacterial propagule: the culture temperature of the staphylococcus aureus, the escherichia coli and the pseudomonas aeruginosa is 30-37 ℃, and the culture time is 3 days; fungi: the culture temperature of the candida albicans is 20-25 ℃, and the culture time is 5 days.

The invention relates to a sterilization rate verification method of an ultraviolet disinfection box, which comprises the steps of treating and sterilizing various culture mediums in a high-temperature high-pressure steam furnace for later use, and preparing 0.9% sodium chloride diluent and eluent; then sterilizing the surface of the object to be in an aseptic state; preparing bacterial suspension, sucking the prepared bacterial suspension to a position on the surface of a sterilized object which is most difficult to sterilize, and preparing bacterial tablets; and then, respectively testing the infected objects with the ultraviolet disinfection box test group and the positive control group to obtain the number of bacteria, and finally judging whether the infected objects are qualified according to the sterilization rate, wherein the sterilization rate verification step is carried out so as to achieve the beneficial effect of accurately and efficiently verifying the sterilization rate of the ultraviolet disinfection box.

Drawings

FIG. 1 is a flow chart of the method for verifying the sterilization rate of the ultraviolet disinfection box.

Detailed Description

The following detailed description of embodiments of the present invention is provided in connection with the accompanying drawings and examples. The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.

Referring to fig. 1, the method for verifying the sterilization rate of an ultraviolet disinfection box disclosed by the invention comprises the following steps:

s1: preparing a sterilization culture medium and a reagent: in this example, the culture media were trypticase Soy peptone broth, Sabouraud's dextrose broth, trypticase Soy peptone agar medium, and Sabouraud's dextrose agar medium, respectively. Treating and sterilizing in a high-temperature high-pressure steam furnace for later use, wherein the temperature of the high-temperature high-pressure steam furnace is set to 121 ℃, and the action time is 20 min;

preparing 0.9% sodium chloride diluent and eluent reagent;

s2: and (3) sterilizing the surface of the object: cleaning and drying an object to be sterilized, sterilizing the object at high temperature and high pressure, and detecting the surface of the object to be sterilized to be in an aseptic state;

s3: preparing a bacterial suspension: taking staphylococcus aureus as an example, preparing a test tube inclined plane of the staphylococcus aureus by a strain passage method, scraping a fresh culture of the staphylococcus aureus by an inoculating loop into a sodium chloride test tube filled with 5mL of 0.9 percent, preliminarily preparing a bacterial suspension, roughly measuring the bacterial concentration of the bacterial suspension with a Mach's turbidimeter, counting the viable bacteria of the bacterial suspension, diluting the bacterial suspension into a bacterial suspension with the bacterial content of 5 multiplied by 105 to 5 multiplied by 106cfu/mL by using a sodium chloride diluent with the concentration, and determining the number of bacterial colonies.

In addition, bacterial suspensions of Escherichia coli, Pseudomonas aeruginosa, Candida albicans can be prepared by the same methods as described above.

S4: preparing a fungus tablet: sucking 0.1mL of bacterial suspension of 5 × 106cfu/mL, dripping 2 × 3cm of the bacterial suspension on the surface of a sterilized object at the position which is most difficult to sterilize, placing the bacterial suspension in a sterilization plate horizontally, placing the sterilization plate and the sterilization plate together in a 37 ℃ thermostat for 30min for drying, dripping a certain amount of bacterial liquid into each carrier, and specifically, the carrier recovery bacterial quantity reaches 5 × 105 cfu/carrier.

Wherein, the inoculum type is respectively bacterial propagule and fungi, and the bacterial propagule comprises: staphylococcus aureus, escherichia coli, pseudomonas aeruginosa; the fungus is Candida albicans.

S5: bacterial count experiments: the experiment was divided into an ultraviolet disinfection box test group and a positive control group, wherein,

the ultraviolet disinfection box test group is as follows: and (4) sterilizing the infected objects in the step S4 in an ultraviolet sterilization box for 60S, then eluting with 100ml of 0.9% sodium chloride reagent, sucking 1ml of the solution into a culture dish, pouring the solution as a plate, inoculating two plates for culture, and counting the plates. Wherein the culture temperature of the staphylococcus aureus, the escherichia coli and the pseudomonas aeruginosa is 30-37 ℃ for 3 days; fungi: the culture temperature of the candida albicans is 20-25 ℃ and the culture time is 5 days.

The positive control group was: directly placing the infected objects in 100ml of 0.9% sodium chloride reagent for elution, sucking 1ml of the eluted objects into a culture dish, pouring the eluted objects as a flat plate, inoculating two flat plates for culture, and counting the number of the flat plates. Similarly, the culture temperature of the staphylococcus aureus, the escherichia coli and the pseudomonas aeruginosa is 30-37 ℃ for 3 days; fungi: the culture temperature of the candida albicans is 20-25 ℃ and the culture time is 5 days.

And performing three parallel experiments in the same group of experiments, testing the results of the three groups of experiments, and taking the lowest sterilization rate value in the three groups as the sterilization result.

S6: and judging the result, according to a calculation formula of the sterilization rate:

and the sterilization rates of the three parallel experiments all reach the target and are judged to be qualified.

In summary, the sterilization rate verification method for the ultraviolet disinfection box of the invention comprises the steps of treating and sterilizing various culture mediums in a high-temperature high-pressure steam furnace for standby, and preparing 0.9% sodium chloride diluent and eluent; then sterilizing the surface of the object to be in an aseptic state; preparing bacterial suspension, sucking the prepared bacterial suspension to a position on the surface of a sterilized object which is most difficult to sterilize, and preparing bacterial tablets; and then, respectively testing the infected objects with the ultraviolet disinfection box test group and the positive control group to obtain the number of bacteria, and finally judging whether the infected objects are qualified according to the sterilization rate, wherein the sterilization rate verification step is carried out so as to achieve the beneficial effect of accurately and efficiently verifying the sterilization rate of the ultraviolet disinfection box.

The above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and substitutions can be made without departing from the technical principle of the present invention, and these modifications and substitutions should also be regarded as the protection scope of the present invention.