CN115228262B - Three-dimensional purification method for acid pollution in laboratory enclosed space - Google Patents
Three-dimensional purification method for acid pollution in laboratory enclosed space Download PDFInfo
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- CN115228262B CN115228262B CN202211011077.XA CN202211011077A CN115228262B CN 115228262 B CN115228262 B CN 115228262B CN 202211011077 A CN202211011077 A CN 202211011077A CN 115228262 B CN115228262 B CN 115228262B
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- 102000039446 nucleic acids Human genes 0.000 claims abstract description 217
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 217
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
- B01D53/34—Chemical or biological purification of waste gases
- B01D53/74—General processes for purification of waste gases; Apparatus or devices specially adapted therefor
- B01D53/77—Liquid phase processes
- B01D53/78—Liquid phase processes with gas-liquid contact
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
- B01D53/30—Controlling by gas-analysis apparatus
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
- B01D53/34—Chemical or biological purification of waste gases
- B01D53/346—Controlling the process
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
- B01D53/34—Chemical or biological purification of waste gases
- B01D53/46—Removing components of defined structure
- B01D53/72—Organic compounds not provided for in groups B01D53/48 - B01D53/70, e.g. hydrocarbons
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
- B01D53/34—Chemical or biological purification of waste gases
- B01D53/74—General processes for purification of waste gases; Apparatus or devices specially adapted therefor
- B01D53/84—Biological processes
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- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F24—HEATING; RANGES; VENTILATING
- F24F—AIR-CONDITIONING; AIR-HUMIDIFICATION; VENTILATION; USE OF AIR CURRENTS FOR SCREENING
- F24F11/00—Control or safety arrangements
- F24F11/30—Control or safety arrangements for purposes related to the operation of the system, e.g. for safety or monitoring
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- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F24—HEATING; RANGES; VENTILATING
- F24F—AIR-CONDITIONING; AIR-HUMIDIFICATION; VENTILATION; USE OF AIR CURRENTS FOR SCREENING
- F24F8/00—Treatment, e.g. purification, of air supplied to human living or working spaces otherwise than by heating, cooling, humidifying or drying
- F24F8/10—Treatment, e.g. purification, of air supplied to human living or working spaces otherwise than by heating, cooling, humidifying or drying by separation, e.g. by filtering
- F24F8/15—Treatment, e.g. purification, of air supplied to human living or working spaces otherwise than by heating, cooling, humidifying or drying by separation, e.g. by filtering by chemical means
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- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
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- F24F—AIR-CONDITIONING; AIR-HUMIDIFICATION; VENTILATION; USE OF AIR CURRENTS FOR SCREENING
- F24F8/00—Treatment, e.g. purification, of air supplied to human living or working spaces otherwise than by heating, cooling, humidifying or drying
- F24F8/10—Treatment, e.g. purification, of air supplied to human living or working spaces otherwise than by heating, cooling, humidifying or drying by separation, e.g. by filtering
- F24F8/175—Treatment, e.g. purification, of air supplied to human living or working spaces otherwise than by heating, cooling, humidifying or drying by separation, e.g. by filtering using biological materials, plants or microorganisms
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- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F24—HEATING; RANGES; VENTILATING
- F24F—AIR-CONDITIONING; AIR-HUMIDIFICATION; VENTILATION; USE OF AIR CURRENTS FOR SCREENING
- F24F8/00—Treatment, e.g. purification, of air supplied to human living or working spaces otherwise than by heating, cooling, humidifying or drying
- F24F8/95—Treatment, e.g. purification, of air supplied to human living or working spaces otherwise than by heating, cooling, humidifying or drying specially adapted for specific purposes
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/20—Air quality improvement or preservation, e.g. vehicle emission control or emission reduction by using catalytic converters
Abstract
The invention provides a three-dimensional purification method for acid pollution in a laboratory enclosed space, which comprises the following steps: s1, space purification: uniformly setting different numbers of air suction points according to the size of the laboratory enclosed space, sucking air in the laboratory enclosed space into the nucleic acid scavenger at the air suction points, controlling the air to stay in the nucleic acid scavenger for at least 3s, so that the air is contacted and mixed with the nucleic acid scavenger and then discharged into the laboratory; s2, verifying the space purifying effect; s3, surface purification; s4, verifying the surface purifying effect; s5, verifying the three-dimensional purifying effect. The method can achieve the three-dimensional purification effect of nucleic acid pollution in the closed space of the laboratory, so as to avoid the occurrence of false positive detection or experimental failure caused by nucleic acid pollution in the laboratory.
Description
Technical Field
The invention belongs to a method for removing nucleic acid, and particularly relates to a three-dimensional purification method for nucleic acid pollution in a laboratory closed space.
Background
Nucleic acid pollution mainly exists in two forms in a closed space, namely, the nucleic acid can be combined with aerosol in the air to form nucleic acid aerosol, namely, the pollution of the nucleic acid in the air usually exists in the form of nucleic acid aerosol; and secondly, nucleic acid is directly attached to a working surface in the closed space, namely, the working surface is directly polluted by the nucleic acid when an operation table, the ground, the surfaces of instruments and equipment are tested.
Aerosols are colloidal dispersions of fine particles of solids and liquids suspended in a gaseous medium and having a particle size of generally 1nm to 1 mm. Nucleic acid aerosol is generated along with the nucleic acid amplification process, and the nucleic acid aerosol pollution can be generated in the processes of air-liquid surface friction, centrifugal separation of a centrifugal machine, violent shaking of a reaction tube, PCR uncapping, repeated sample suction of a liquid transfer device, leakage of pollutants and the like. Once formed, the nucleic acid aerosol is not only suspended in the gaseous medium for a long period of time, but also partially settles on the solid phase surfaces of laboratory desktops, instruments, consumables, floors, etc. with the circulation of air. Thus, in the closed space, the contamination of the nucleic acid is dynamically changed, and it is difficult to ensure that the nucleic acid in the closed space is thoroughly purged by conventional purging means.
The molecular laboratory is used as a scientific research and clinical examination place, the use frequency of PCR is high, the condition of repeated treatment of samples and amplification targets in batches often occurs, the nucleic acid pollution is continuously accumulated in an experiment area, and the pollution risk is continuously increased. With the continuous improvement of amplification efficiency and sensitivity of PCR and other technologies, the occurrence frequency of false positives of detection results is increased more and more due to the generated nucleic acid pollution. False positives mean that the experimental results are not reliable and directly result in a loss of laboratory economy. Even more serious, once nucleic acid contamination has formed, contamination of the entire PCR laboratory can occur and is difficult to clean thoroughly in a short period of time, even requiring the laboratory to be shut down for a period of time if necessary.
Disclosure of Invention
Nucleic acid is directly attached to the working surface to cause nucleic acid pollution of the working surface, and meanwhile, the nucleic acid pollutes the air in a form of nucleic acid aerosol, in addition, the nucleic acid aerosol naturally settles to the working surface, and the nucleic acid aerosol or the nucleic acid pollution of the working surface is also diffused into the air to form the nucleic acid aerosol pollution, so that the nucleic acid pollution in the closed space is always a dynamic complex process; at present, the sterilization of the closed space of a laboratory generally has better effect on bacteria, viruses and the like, and no systematic method for nucleic acid pollution can ensure that the nucleic acid in the closed space is thoroughly purified; aiming at the problems, the invention provides a three-dimensional purification method for nucleic acid pollution in a laboratory enclosed space, which can ensure the three-dimensional purification effect of the nucleic acid pollution in the laboratory enclosed space so as to avoid the occurrence of false positive detection or experimental failure caused by nucleic acid pollution in the laboratory.
The invention is realized by the following technical scheme.
The three-dimensional purification method for the nuclear acid pollution in the closed space of the laboratory is characterized by comprising the following steps:
s1, space purification: according to the size of the laboratory enclosed space, different numbers of air suction points are uniformly arranged, air in the laboratory enclosed space is sucked into the nucleic acid scavenger at the air suction points, the air is controlled to stay in the nucleic acid scavenger at least for 3 s, and the air is discharged into the laboratory after being contacted and mixed with the nucleic acid scavenger;
In the step, air with nucleic acid aerosol pollution is fully contacted, mixed and reacted with the nucleic acid scavenger by sucking air with different air suction points in a laboratory closed space into the nucleic acid scavenger, so that nucleic acid is degraded, and purified air can be discharged, so that the effect of purifying the laboratory closed space is achieved;
s2, verifying the space purifying effect: after the space purification in the step S1 is finished, detecting the content of residual nucleic acid in the air of each air suction point in the closed space respectively, if the detection results are negative, entering the step S3, and if the detection results are positive, repeating the steps S1 and S2 until the detection results of all the air suction points in the step S2 are negative;
because the nucleic acid aerosol in the air is suspended in the air for a long time, the weight and the grain size are probably increased continuously, and finally, part of the nucleic acid aerosol continuously and slowly subsides on the working surface, so that the working surface is polluted; while the sedimentation pollution of the nucleic acid aerosol cannot be controlled, the sedimentation pollution of the nucleic acid aerosol from the subsequent space to the working surface can be avoided only after the nucleic acid aerosol in the air is thoroughly purified by the space purification in the step S1; otherwise, if the space is not thoroughly purified, the nucleic acid aerosol in the space still settles to pollute the surface after the surface is thoroughly purified in the step S3; therefore, after the space purification of step S1 is completed, it is necessary to detect the content of the residual nucleic acid in the air at each air suction point in the closed space to ensure the effect of the space purification of step S1;
S3, surface purification: spraying the nucleic acid scavenger to a working surface in a laboratory closed space in a spray mode, so that the nucleic acid scavenger uniformly wets the working surface, and naturally airing the nucleic acid scavenger on the working surface;
in the experimental operation process, the nucleic acid is likely to be directly stained on the working surface, or the nucleic acid aerosol suspended in the air subsides to pollute the working surface, so that the working surface in the closed space of the laboratory is polluted by the nucleic acid aerosol; by spraying the nucleic acid scavenger in the form of a spray onto the working surface in the laboratory enclosed space, the nucleic acid scavenger can be effectively contacted with the working surface under the condition of reducing the use amount of the nucleic acid scavenger, so that the nucleic acid aerosol settled on the working surface is degraded;
s4, verifying the surface purification effect: immediately sampling and detecting the content of the residual nucleic acid of the working surface near each air suction point after the surface cleaning treatment in the step S3, wherein the number of the sampling points of the working surface near each air suction point is 10-15, and the 10-15 sampling points are distributed on at least 3 different working surfaces near the air suction point; if the detection results are negative, the step S5 is carried out, and if the positive result is detected, the step S3 and the step S4 are repeatedly circulated until the detection results of the step S4 are negative;
After the surface is purified, the step S3 of sampling and verifying whether the surface is thoroughly purified is needed immediately; if the surface purification in step S3 is not thorough, it is possible that some nucleic acid contamination of the working surface still exists, and in the subsequent detection or experimental operation, along with the movement of personnel and unavoidable touching of the working surface, there is still a certain risk that the nucleic acid contamination remaining on the working surface is resuspended in air along with the flow of air, so that the three-dimensional purification of the laboratory enclosed space cannot be realized, and therefore it is necessary to verify whether the surface purification is thorough through step S4;
s5, verifying the three-dimensional purification effect: after the detection results in the verification of the surface purification effect in the step S4 are negative, 2-4 h are obtained, and the content of the residual nucleic acid on the working surface near each air suction point is immediately sampled and detected, wherein the number of sampling points of the working surface near each air suction point is 20-30, and the 20-30 sampling points are distributed on at least 3 different working surfaces near the air suction point; if all the detection results are negative, the three-dimensional purification of the acid pollution in the closed space is finished; if the positive result is detected, repeating the steps S1 to S5 until the detection results of the step S5 are all negative.
Since steps S1 to S4 are sequentially completed, however, the remaining nucleic acid aerosol in the air will be settled to pollute the working surface, and the remaining nucleic acid aerosol or the nucleic acid pollution on the working surface will pollute the air along with the movement of the personnel or the touch with the working surface, the following situations may exist: when the surface purification is performed in the step S3, as personnel need to perform necessary operations and touch the working surface, nucleic acid pollution on the working surface is resuspended in air along with the flow of air in the surface purification process and is difficult to be totally settled to the working surface in a short time, so that the verification results of the surface purification effect in the subsequent step S4 are negative, that is, although the verification results of the space purification effect in the step S2 and the verification results of the surface purification effect in the step S4 are both negative, the three-dimensional purification of the nucleic acid pollution in the closed space of a laboratory still cannot be ensured to be thorough, namely the situation of false negative purification results is possible; in contrast, after the applicant carries out the surface purification for the last cycle treatment for 2-4 hours in the step S3, sampling and detecting the content of the residual nucleic acid on the working surface near different suction points, if the air is polluted again in the step S4, sampling the working surface after the detection results in the step S4 are negative and 2-4 h, and detecting positive results in the step S5 certainly because of sedimentation pollution of the nucleic acid aerosol; if the detection results in the step S5 are negative, the laboratory closed space is completely purified in a three-dimensional way;
In addition, regarding the conditions and timing of the three-dimensional purification effect verification in step S5, the applicant has the following consideration: if the positive result is detected in the surface purification effect verification in the step S4, the surface purification in the laboratory closed space is not thorough, namely the nucleic acid pollution of certain working surfaces still exists, and in order to save unnecessary experimental links and save purification time, the step S5 is unnecessary; and only when the detection results in the verification of the surface cleaning effect in the step S4 are all negative, it is necessary to verify whether the above-described false negative cleaning result exists or not by the step S5. In order to judge whether the nucleic acid aerosol pollution exists in the air or not at the detection time, in the step S2, a natural sedimentation method for the nucleic acid aerosol is adopted for sampling, namely, a blank culture dish is placed in a space to be detected for a period of time, the nucleic acid aerosol is naturally sedimented into the culture dish to collect the nucleic acid aerosol for detection, the preferred range of sedimentation time (the placement time of the culture dish) is discussed in the embodiment, and the result shows that the sedimentation time is 15-20 min, so that the nucleic acid aerosol pollution in the air can be ensured to be detected; however, since the verification of step S5 is to determine whether or not there is a false negative result, for which the nucleic acid aerosol in the air may be less contaminated because it is "nucleic acid contamination on the work surface is resuspended in the air with the flow of air during the surface purification and is difficult to be totally settled to the work surface in a short time", if the nucleic acid aerosol is collected by natural settling as well, the settling time should be increased so that the nucleic acid aerosol diffused into the air has a sufficient settling time to settle to the work surface. In this regard, the applicant comprehensively considers the above factors, and selects 2 to 4 h to sample the working surface after the detection results in the step S4 are negative, that is, if the nucleic acid aerosol pollution exists in the air, the nucleic acid aerosol is settled for 2 to 4 h (far more than 15 to 20 minutes), thereby being beneficial to collecting the effective nucleic acid pollution from the working surface; in addition, the applicant increases the number of sampling points in step S5, and controls the number of sampling points of the working surface near each air suction point to be 20-30, so as to ensure that the nucleic acid pollution in the air under the false negative purification result can be detected.
Preferably, in the step S1, the nucleic acid scavenger is a universal nuclease solution (UNC enzyme solution) or hypochlorous acid solution; in the step S3, the nucleic acid scavenger is a universal nuclease solution (UNC enzyme solution) or hypochlorous acid solution.
Preferably, in the step S2, the number of sampling points of each air sucking point is 10-15, and the 10-15 sampling points should be distributed in at least 3 different spatial positions near the air sucking point.
Preferably, in the step S2, the sampling manner of the sampling points is as follows: and (3) placing a blank culture dish at a sampling point, naturally settling the nucleic acid aerosol in the air for 15-20 min into the blank culture dish, and further detecting whether nucleic acid pollution exists in the air or not according to whether the nucleic acid pollution exists in the culture dish or not.
Preferably, in the step S2, the number of sampling points of each air sucking point is 10-15, and the 10-15 sampling points should be distributed in at least 3 different spatial positions near the air sucking point.
Preferentially, in the step S1, the air suction points synchronously suck the air in the laboratory closed space at the same air suction rate, so that the air in the space can be uniformly and effectively sucked and contacted and mixed with the nucleic acid scavenger, and the efficiency of space purification is improved.
Preferably, in the step S1, every 40 to 100 m of the laboratory enclosed space 3 An air suction point is provided.
Preferably, in the step S1, the total suction amount of the air is controlled to be 2-5 times of the laboratory enclosed space.
Preferably, in the step S1, the nucleic acid scavenger is usedThe volume ratio of the product to the laboratory closed space is 1-5:100L/m 3 。
Preferably, the working surface is the outer surface of facilities, equipment and vessels used for experimental operation in a laboratory closed space and the outer surface of clothes of operators.
Preferably, in the step S4 and the step S5, a high pollution area of the working surface is sampled by a cotton swab wiping standard sampling method, so as to detect the content of the nucleic acid remained on the working surface.
Preferentially, the sampling area of the high pollution area of the working surface is 3-8 cm multiplied by 3-8 cm; the high pollution area of the working surface comprises the outer surfaces of an experiment operation table top, instrument equipment, a test tube rack, a mobile phone shell, a refrigerator, an ultra-clean workbench, a door handle, a stool, the ground and an illumination switch.
Preferably, the nucleic acid content is detected by fluorescent quantitative PCR.
1) According to the invention, the nucleic acid pollution in the air in the laboratory closed space can be effectively purified through space purification, and the nucleic acid pollution on the working surface in the laboratory can be effectively purified through surface purification, so that the three-dimensional purification of the laboratory space and the surface is realized.
2) Since sedimentation pollution of the nucleic acid aerosol in the air is unavoidable, the risk of recontamination of the working surface by the nucleic acid aerosol in the air is high; while surface diffusion pollution only occurs with a certain probability, the risk of diffusing polluted air is low. Therefore, aiming at the characteristics that the nucleic acid aerosol has high risk of air sedimentation pollution and low risk of surface diffusion pollution, the invention firstly eliminates the high risk of nucleic acid aerosol sedimentation pollution through space purification, performs effect verification, then performs surface purification to eliminate low risk of nucleic acid diffusion pollution, simultaneously performs effect verification, and through scientific control of the purification and verification steps, the invention gradually goes through, and finally, through three-dimensional purification effect verification, the three-dimensional purification of the nucleic acid pollution in the closed space of a laboratory can be ensured to be thorough, so that the condition that the laboratory detects false positive or experimental failure due to nucleic acid pollution is avoided.
Drawings
FIG. 1 is a flow chart of the method of the present invention.
Description of the embodiments
The following description of the embodiments of the present invention will be apparent and complete, and it is to be understood that the embodiments described are merely some, but not all embodiments of the present invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
Referring to fig. 1, the method for three-dimensional purification of acid pollution in a laboratory enclosed space comprises the following steps:
s1, space purification
According to the size of the laboratory enclosed space, different numbers of air suction points are uniformly arranged, air in the laboratory enclosed space is sucked into the nucleic acid scavenger at the air suction points, the air is controlled to stay in the nucleic acid scavenger at least for 3 s, and the air is discharged into the laboratory after being contacted and mixed with the nucleic acid scavenger;
s2, verifying space purifying effect
After the space purification in the step S1 is finished, detecting the content of residual nucleic acid in the air of each air suction point in the closed space respectively, if the detection results are negative, entering the step S3, and if the detection results are positive, repeating the steps S1 and S2 until the detection results of all the air suction points in the step S2 are negative;
s3, surface purification
Spraying the nucleic acid scavenger to a working surface in a laboratory closed space in a spray mode, so that the nucleic acid scavenger uniformly wets the working surface, and naturally airing the nucleic acid scavenger on the working surface;
s4, verifying surface purification effect
Immediately sampling and detecting the content of the residual nucleic acid of the working surface near each air suction point after the surface cleaning treatment in the step S3, wherein the number of the sampling points of the working surface near each air suction point is 10-15, and the 10-15 sampling points are distributed on at least 3 different working surfaces near the air suction point; if the detection results are negative, the step S5 is carried out, and if the positive result is detected, the step S3 and the step S4 are repeatedly circulated until the detection results of the step S4 are negative;
S5, three-dimensional purification effect verification
Immediately sampling and detecting the content of the residual nucleic acid of the working surface near each air suction point after the detection results in the step S4 are all negative and 2-4 h, wherein the number of sampling points of the working surface near each air suction point is 20-30, and the 20-30 sampling points are distributed on at least 3 different working surfaces near the air suction point; if all the detection results are negative, the three-dimensional purification of the acid pollution in the closed space is finished; if the positive result is detected, repeating the steps S1 to S5 until the detection results of the step S5 are all negative.
Example 2
Referring to fig. 1, the method for three-dimensional purification of acid pollution in a laboratory enclosed space comprises the following steps:
s1, space purification
According to the size of the laboratory enclosed space, different numbers of air suction points are uniformly arranged, air in the laboratory enclosed space is sucked into the nucleic acid scavenger at the air suction points, the air is controlled to stay in the nucleic acid scavenger at least for 3 s, and the air is discharged into the laboratory after being contacted and mixed with the nucleic acid scavenger; wherein, different numbers of air suction points are uniformly arranged according to the size of the laboratory enclosed space, namely, every 40 to 100 m of the laboratory enclosed space 3 Setting an air suction point, synchronously sucking air in a laboratory enclosed space by adopting the same air suction rate at different air suction points, controlling the total sucking amount of the air to be 2-5 times of the laboratory enclosed space, and controlling the volume ratio of the nucleic acid scavenger to the laboratory enclosed space to be 1-5:100L/m 3 The method comprises the steps of carrying out a first treatment on the surface of the The nucleic acid scavenger is UNC enzyme solution or hypochlorous acid solution; when the UNC enzyme solution is used as a nucleic acid scavenger, the concentration of the UNC enzyme is preferably 3-4 ug/mL; when a hypochlorous acid solution is used as a nucleic acid scavenger, the hypochlorous acid solution is preferably used at a concentration of 400～600 ppm；
S2, verifying space purifying effect
After the space purification in the step S1 is finished, respectively detecting the content of residual nucleic acid in the air at each air suction point in the closed space by adopting fluorescent quantitative PCR, if the detection results are negative, entering the step S3, and if the detection results are positive, repeating the steps S1 and S2 until the detection results in the step S2 are negative; wherein the number of sampling points of each air suction point is 10-15, and the 10-15 sampling points are distributed at least 3 different spatial positions near the air suction point; placing a blank culture dish at a sampling point, naturally settling the nucleic acid aerosol in the air for 15-20 min into the blank culture dish, and further representing whether nucleic acid pollution exists in the air or not according to the nucleic acid content in the culture dish;
S3, surface purification
If the UNC enzyme solution is used as a nucleic acid scavenger, the concentration of the UNC enzyme is preferably 3-4 ug/mL, spraying the UNC enzyme solution to a working surface in a laboratory closed space in a spraying mode, so that the UNC enzyme solution uniformly wets the working surface, forming a layer of water film on the working surface, and naturally airing the UNC enzyme solution on the working surface;
if hypochlorous acid solution is used as a nucleic acid scavenger, the recommended use concentration of the hypochlorous acid solution is 400-600 ppm, the hypochlorous acid solution is sprayed to the working surface in a laboratory closed space in a spraying mode, so that the hypochlorous acid solution wets the working surface uniformly, after the hypochlorous acid solution is naturally dried on the working surface, pure water is required to wipe the working surface, and residues of the hypochlorous acid solution are timely wiped off, so that the hypochlorous acid solution is prevented from being corroded and polluted for a long time;
the working surface is the outer surface of equipment, equipment and utensils used for experimental operation in the laboratory closed space and the outer surface of clothes of operators;
s4, verifying surface purification effect
Immediately sampling and detecting the content of the residual nucleic acid of the working surface near each air suction point after the surface cleaning treatment in the step S3, wherein the number of the sampling points of the working surface near each air suction point is 10-15, and the 10-15 sampling points are distributed on at least 3 different working surfaces near the air suction point; if the detection results are negative, the step S5 is carried out, and if the positive result is detected, the step S3 and the step S4 are repeatedly circulated until the detection results of the step S4 are negative; sampling a high-pollution area of the working surface near each air suction point by adopting a cotton swab wiping standard sampling method to detect the content of residual nucleic acid in the working surface, wherein the sampling area of the high-pollution area of the working surface is 3-8 cm multiplied by 3-8 cm, and the high-pollution area of the working surface comprises the outer surfaces of an experiment operation table top, instrument equipment, a test tube rack, a liquid transfer device, a mobile phone shell, a refrigerator, an ultra-clean workbench, a door handle, a stool, the ground and a lighting switch;
S5, three-dimensional purification effect verification
Immediately sampling and detecting the content of the residual nucleic acid of the working surface near each air suction point after the detection results in the step S4 are all negative and 2-4 h, wherein the number of sampling points of the working surface near each air suction point is 20-30, and the 20-30 sampling points are distributed on at least 3 different working surfaces near the air suction point; if all the detection results are negative, the three-dimensional purification of the acid pollution in the closed space is finished; if the positive result is detected, repeating the steps S1 to S5 until the detection results of the step S5 are negative; the high pollution area of the working surface near each air suction point is sampled by adopting a cotton swab wiping standard sampling method to detect the content of the residual nucleic acid of the working surface, the sampling area of the high pollution area of the working surface is 3-8 cm multiplied by 3-8 cm, and the high pollution area of the working surface comprises the outer surfaces of an experimental operation table top, instrument equipment, a test tube rack, a liquid transfer device, a mobile phone shell, a refrigerator, an ultra-clean workbench, a door handle, a stool, the ground and an illumination switch.
Example 3
Influence of different sedimentation time on detection results of nucleic acid aerosol natural sedimentation method sampling.
1. Principle of natural sedimentation method of nucleic acid aerosol: by utilizing the characteristic that nucleic acid aerosol in air continuously and naturally subsides, nucleic acid subsides to the culture dish from the air is collected through a blank culture dish placed in the air for a period of time (subsidence time), and whether nucleic acid aerosol pollution exists in the air is characterized by whether nucleic acid is detected in the culture dish, if the nucleic acid is detected in the blank culture dish, the nucleic acid pollution exists in the air, otherwise, the nucleic acid pollution does not exist in the air.
2. The detection method comprises the following steps: the prepared 300 mL PCR fragment test solution with Ct value of 20.6 was subjected to space size 79 m by a CN61 aerosol generator 3 Spraying treatment is carried out in a left closed laboratory and a right closed laboratory, then 9 blank culture dishes (the specifications of the culture dishes are 150 mm, the culture dishes are strictly disinfected and have no nucleic acid pollution) are randomly placed in the closed laboratory, after 15min, 20min and 30min of the culture dishes are sequentially placed, any 3 culture dishes are taken out of the 9 culture dishes respectively, samples are immediately collected by cotton swabs and eluted in 1 mL sterile water, and fluorescent quantitative PCR detection is carried out on eluent to investigate the change of the nucleic acid content detected in the culture dishes under different sedimentation time, and the results are shown in table 1; in the invention, the manufacturer of the CN61 aerosol generator is Collison company in the United states; the model of the real-time fluorescence quantitative PCR instrument is SLAN-96S of a full-automatic medical PCR analysis system;
TABLE 1 detection results of nucleic acid residues in air
As can be seen from Table 1, the nucleic acid pollution source was sprayed into the closed space and allowed to naturally settle, and the settled nucleic acid was collected by a petri dish, and the detection results at different settling times showed that the longer the settling time, the smaller the Ct value, and the higher the detected nucleic acid content. The sedimentation is carried out for 15 min, compared with the sedimentation for 20 min, the Ct value is reduced, the reduction amplitude is not large, and the sedimentation results for 20 min and sedimentation results for 30min are not significantly different, but nucleic acid can be detected in the culture dish after sedimentation for 15 min-30 min, so that in order to save detection time, the nucleic acid pollution in the air can be detected, and 15 min-20 min can be selected as sedimentation time to detect the nucleic acid pollution in the air.
Example 4
The effect of space purification in the method of the invention is verified.
Selecting the size of the two spaces to be 79 m 3 The left and right closed laboratories are used as processing objects, two bottles of PCR fragment test solution 300 mL with Ct value about 20 are respectively and simultaneously sprayed into the closed spaces of the two laboratories and the surfaces of equipment and facilities in a spraying mode by using a CN61 aerosol generator, one of the two laboratories is used as a comparison group, and the other laboratory is used as a test group;
The experimental group adopts the step S1 in the method for space purification, while the comparison group does not adopt any measure; specifically, the method for purifying the space comprises the following steps:
the 3L nucleic acid scavenger (nucleic acid scavenger is UNC enzyme solution with concentration of UNC enzyme of 3.4 ug/mL, using 25mm Tris-HCl (pH=8.2), 10mm NaCl,2mm MgCl 2 Buffer solution with concentration is configured) to be placed in a container, wherein the mouth of the container is selected to be narrow and the bottom depth is selected so as to ensure that the sucked air has sufficient contact time with the nucleic acid scavenger; since the laboratory enclosed space of the experimental group has a volume of 79 m 3 The left and right sides are provided with only one air suction point, and the air suction point is positioned in the middle of the laboratory closed space of the experimental group as much as possible; pumping air into the bottom of the container at the air pumping point by using a pump body to ensure that the pumped air stays at least 3 s in the nucleic acid scavenger and is automatically discharged; in order to ensure that the polluted air in the laboratory enclosed space of the experimental group can be pumped into the container as much as possible, the total suction amount of the air is controlled to be 3 times that of the laboratory enclosed space, and under the condition that the suction efficiency of the pump body and the volume of the laboratory enclosed space of the experimental group are determined, only the suction time is required to be reasonably controlled, and the space purification time set in the embodiment is 60 minutes.
After the space purification of the experimental group is completed, the content of the residual nucleic acid in the air at the air suction point of the experimental group is detected by adopting fluorescent quantitative PCR, and meanwhile, the content of the residual nucleic acid in the air of the comparison group is detected by adopting fluorescent quantitative PCR, and the specific process is as follows:
for the experimental group, 10 blank culture dishes (the specification of the culture dishes is 150 mm, the culture dishes are strictly disinfected and have no nucleic acid pollution) are randomly placed on the ground on an experimental operation table near the suction point immediately after the space purification of the experimental group is completed, the sampling points are distributed at different space positions of the air suction point, after the culture dishes are kept stand for 20 min, the culture dishes are taken out, samples are collected by using cotton swabs and eluted in 1 mL sterile water, and fluorescent quantitative PCR detection is carried out on eluent, so that the results are shown in Table 2;
for the comparison group, the same sampling method, sampling time and detection mode as those of the experimental group are adopted, and the results are shown in table 2;
TABLE 2 Effect detection of spatial purification
As can be seen from Table 2, the PCR fragments could not be detected in the blank culture dish after the space purification, so that the nucleic acid contamination in the air in the laboratory enclosed space of the experimental group can be thoroughly purified by the space purification in the invention. Generally, in the invention, the space purification of the step S1 can effectively purify the nucleic acid pollution in the air in the closed space of the laboratory, but the purification is required to be completed by experimenters through standard operation under the set parameter condition, so that the condition that the space purification is not thorough due to the existence of objective factors of artificial reasons or accidents is unavoidable, and therefore, the step S2 space purification effect verification is required to be carried out after the step S1 space purification is carried out so as to judge whether the nucleic acid in the air is thoroughly purified.
Example 5
The effect of surface purification in the method of the invention is verified.
Selecting 5 cm ×5 cm areas on different working surfaces of a laboratory, spraying 100 uL PCR fragment test solution (Ct value of about 20) on the different working surfaces, naturally airing, wiping the whole surface with a sterile cotton swab, eluting in 1mL sterile water, performing fluorescent PCR detection on the eluent, and determining the nucleic acid pollution condition of the surface before purification, wherein the result is shown in Table 2;
according to the method of the step S3, namely 0.4 mL hypochlorous acid solution is sprayed to the area of 5 cm multiplied by 5 cm of the different working surfaces in a spraying mode, so that the working surfaces are uniformly wetted by the hypochlorous acid solution, the hypochlorous acid solution is naturally dried on the working surfaces (the drying time after spraying is controlled to be 30 min), then the whole surface is wiped by a sterile cotton swab, the whole surface is sampled and eluted in 1mL of sterile water, the eluent is subjected to fluorescent PCR detection, and the nucleic acid pollution condition of the purified surface is measured, and the result is shown in Table 3;
TABLE 3 Effect detection of surface purification
As shown in the results of Table 3, the sampling detection results of the working face are negative after the surface purification by adopting the method provided by the invention, which shows that the method has a better effect of removing the nucleic acid pollution on the solid surface. However, the surface purification in step S3 of the present invention needs to be performed by experimenters through standard operations under the set parameter conditions, so that there is inevitably an objective factor of artificial reasons or accidents to cause incomplete space purification, such as uneven spraying on the working surface, or that some working surfaces are not sprayed, so that the surface purification effect verification in step S4 needs to be performed after the surface purification in step S3 to determine whether the nucleic acid pollution on the working surface is thoroughly purified.
Example 6
The size of the selected space is 79 m 3 The laboratory was treated, and the prepared 300 mL PCR fragment test solution with Ct value of about 20 was sprayed into the closed space and on the working surface of the laboratory by means of CN61 aerosol generator, so that the laboratory was contaminated with nucleic acid, and then the laboratory of the laboratory group was subjected to three-dimensional purification by the method of the present invention, please refer to fig. 1, the specific procedure is as follows:
s1, space purification
Nucleic acid scavengersAs the UNC enzyme solution, the concentration of UNC enzyme in the UNC enzyme solution was 3.4. 3.4 ug/mL, 25mm Tris-HCl (pH=8.2) was used, 10mm NaCl,2mm MgCl 2 Preparing a buffer solution with concentration; placing 3L nucleic acid scavenger in a container, wherein the container is selected to be narrow in mouth and deep in bottom so as to ensure sufficient contact time between the sucked air and the nucleic acid scavenger; since the laboratory enclosed space of the experimental group has a volume of 79 m 3 The left and right sides are provided with only one air suction point, and the air suction point is positioned in the middle of the laboratory closed space of the experimental group as much as possible; pumping air into the bottom of the container at the air pumping point by using a pump body to ensure that the pumped air stays at least 3 s in the nucleic acid scavenger and is automatically discharged; in the air pumping-in and discharging process, nucleic acid aerosol carried in the air is contacted, mixed and reacted with a nucleic acid scavenger, so that nucleic acid in the nucleic acid aerosol is degraded, and finally purified air can be discharged, thereby achieving the effect of purifying the laboratory closed space of the laboratory; in order to ensure that the polluted air in the laboratory enclosed space of the experimental group can be pumped into the container as much as possible, the total suction amount of the air is controlled to be 2-5 times of that of the laboratory enclosed space, and under the condition that the suction efficiency of the pump body and the volume of the laboratory enclosed space of the experimental group are determined, only reasonable control of the suction time is needed, and the space purification time is set to be 60 minutes in the embodiment.
S2, verifying space purifying effect
After the space purification in the step S1 is completed, the content of residual nucleic acid in the air at the air suction point is detected by adopting fluorescence quantitative PCR, and the specific process is as follows:
after the space purification in the step S1 is completed, 10 blank culture dishes (the specifications of the culture dishes are 150 mm, the culture dishes are strictly sterilized and have no nucleic acid pollution, 4 culture dishes are placed on a table of 10 blank culture dishes, the number is 1-4, 4 culture dishes are placed on the ground, the number is 5-8, 2 culture dishes are placed on the stool, the number is 9-10) are placed on the experiment operation table, the ground and the stool near the suction point, sampling points are distributed at different space positions near the air suction point, after the culture dishes stand for 20min, the culture dishes are taken out, a cotton swab collected sample is eluted in 1 mL sterile water, and an eluent is taken for fluorescence quantitative PCR detection, and the result is shown in a table 4;
TABLE 4 detection results of nucleic acid residues in laboratory air after spatial purification
As can be seen from Table 4, after the space purification in step S1, the PCR fragment could not be detected in the blank petri dish settled for 20min, and thus it can be seen that the purification of nucleic acid contamination in the air in the closed space of the laboratory group was completely completed in step S1, and the process could proceed to step S3.
S3, surface purification
The nucleic acid scavenger is UNC enzyme solution with concentration of 3.4 ug/mL UNC enzyme, 25mm Tris-HCl (pH=8.2) is used, 10mm NaCl,2mm MgCl 2 Preparing a buffer solution with concentration; spraying the nucleic acid scavenger to a working surface in a laboratory closed space in a spray mode, so that the nucleic acid scavenger wets the working surface uniformly, forming a layer of water film on the working surface, and naturally airing the nucleic acid scavenger on the working surface; the working surface is the outer surface of equipment, equipment and utensils used for experimental operation in the laboratory closed space and the outer surface of clothes of operators;
s4, verifying surface purification effect
Immediately after the surface cleaning treatment in step S3, the remaining nucleic acid content of the working face in the vicinity of the air suction point was detected by fluorescent quantitative PCR sampling, wherein the number of sampling points of the working face in the vicinity of the air suction point was 10, and the 10 sampling points were distributed on the following 6 working faces in the vicinity of the air suction point: ground (3 sampling points), refrigerator (2 sampling points), door handle (1 sampling point), ep pipe rack (1 sampling point), stool (1 sampling point) and experiment operation table (2 sampling points); for the sampling point, a cotton swab wiping standard sampling method is adopted in sampling, namely, a surface with a sterile cotton swab wiping area of 5 cm multiplied by 5 cm is eluted in 1mL of sterile water, and fluorescent PCR detection is carried out on the eluent, wherein the detection result is as follows:
TABLE 5 detection results of nucleic acid residues on working surfaces in laboratory after surface purification
As shown in table 5, if the detection results of 10 sampling points of 6 different working surfaces are all negative, step S5 is entered to perform three-dimensional purification effect verification;
s5, three-dimensional purification effect verification
Immediately after the detection results in step S4 are negative, 2 h, detecting the content of the nucleic acid remained on the working surface near the air suction point by using fluorescence quantitative PCR, wherein the number of the sampling points on the working surface near the air suction point is 20, and the 20 sampling points are distributed on the following 6 working surfaces near the air suction point: ground (8 sampling points), refrigerator (2 sampling points), door handle (1 sampling point), ep pipe rack (1 sampling point), stool (2 sampling points) and experiment operation table (6 sampling points); for the sampling point, a cotton swab wiping standard sampling method is adopted in sampling, namely, a surface of a working surface with the area of 5 cm multiplied by 5 cm is wiped by a sterile cotton swab and eluted in 1mL of sterile water, and fluorescent PCR detection is carried out on eluent, wherein the detection result is as follows:
table 6 test results of three-dimensional purification effect verification
As shown in Table 6, the detection results of the total 20 sampling points of 6 different working surfaces are negative, and the fact that no false negative purification result exists indicates that the nucleic acid pollution in the closed space of the laboratory is thoroughly purified.
Example 7
The size of the selected space is 150 m 3 The laboratory with left and right rectangular house type is used as a processing object, the prepared PCR fragment test solution with Ct value of about 20 and 800 mL is sprayed into the closed space and the working surface of the laboratory in a spray mode by using a CN61 aerosol generator, so that the laboratory forms nucleic acid pollution, and then the method is adoptedThe laboratory of the experimental group was purified three-dimensionally by the method, see fig. 1, and the specific process is as follows:
s1, space purification
The nucleic acid scavenger is UNC enzyme solution with concentration of 3.4 ug/mL UNC enzyme, 25mm Tris-HCl (pH=8.2) is used, 10mm NaCl,2mm MgCl 2 Preparing a buffer solution with concentration; dividing the 8L nucleic acid scavenger into two parts, respectively placing the two parts in two independent containers, wherein the selected containers are narrow in mouth and deep in bottom so as to ensure that the sucked air has sufficient contact time with the nucleic acid scavenger; since the laboratory enclosed space of the experimental group has a volume of 150 m 3 The left and right sides are provided with only two air suction points, namely a No. 1 air suction point and a No. 2 air suction point, which are respectively positioned at the middle positions of the left and right sides of the laboratory enclosed space of the experimental group; pumping air into the bottom of the container filled with the 4L nucleic acid scavenger by using two pump bodies at the two air pumping points respectively, so that the pumped air stays at least 3 s in the nucleic acid scavenger, and then is automatically discharged; in the air pumping-in and discharging process, nucleic acid aerosol carried in the air is contacted, mixed and reacted with a nucleic acid scavenger, so that nucleic acid in the nucleic acid aerosol is degraded, and finally purified air can be discharged, thereby achieving the effect of purifying the laboratory closed space of the laboratory; in order to ensure that the polluted air in the laboratory enclosed space of the experimental group can be pumped into the container as much as possible, the total suction amount of the air is controlled to be 2-5 times of that of the laboratory enclosed space, and under the condition that the suction efficiency of the pump body and the volume of the laboratory enclosed space of the experimental group are determined, only reasonable control of the suction time is needed, and the space purification time is set to be 60 minutes in the embodiment.
S2, verifying space purifying effect
After the space purification in the step S1 is completed, the content of residual nucleic acid in the air at the air suction point is detected by adopting fluorescence quantitative PCR, and the specific process is as follows:
after the space purification in the step S1 is finished, 10 blank culture dishes (the culture dishes are 150 mm in specification and have no nucleic acid pollution after strict disinfection) are respectively placed on an experiment operation table, the ground and a stool near the No. 1 air suction point and the No. 2 air suction point, so that sampling points are distributed at different space positions near the air suction point; 4 blank culture dishes with the number of 1-4 are placed on a table of 10 blank culture dishes with the number of 1-4; 4, 5-8 numbered, are placed on the ground; 2 stools are placed on the bench with the number of 9-10; 4 blank culture dishes with the same number 2 air suction points are placed on a table, and the numbers are 11-14; 4, 15-18, are placed on the ground; 2 stools are placed on the bench with the number of 19-20; standing the blank culture dish for 20 min, and taking out the culture dish; then the blank culture dish is eluted in 1 mL sterile water by using a cotton swab collection sample, and the eluent is taken for fluorescence quantitative PCR detection, and the result is shown in Table 7;
TABLE 7 detection results of nucleic acid residues in laboratory air after spatial purification
As is clear from Table 7, after the space purification for 60 min and 120 min, the PCR fragment was detected at the air suction point 2, that is, a positive detection was found, and it was found that the complete purification of nucleic acid in the air in the laboratory enclosed space was not achieved in the step S1, and therefore, the space purification was required to be repeated again in the step S1. The method for verifying the spatial purification effect after repeated purification is not repeated here, and only verification results are listed, specifically as follows:
TABLE 8 detection results of nucleic acid residues in laboratory air after spatial purification
As shown in Table 8, after repeating step S1, the PCR fragments could not be detected in the blank culture dish after the second space purification, so that the purification of nucleic acid contamination in the air in the laboratory enclosed space of the laboratory was completely completed, and the process could proceed to step S3.
S3, surface purification
The hypochlorous acid solution is used as a nucleic acid scavenger, the recommended use concentration of the hypochlorous acid solution is 500 ppm, the hypochlorous acid solution is sprayed to the working surface in a laboratory closed space in a spraying mode, the working surface is uniformly wetted by the hypochlorous acid solution, after the hypochlorous acid solution is naturally dried on the working surface, the working surface is required to be wiped by pure water, and residues of the hypochlorous acid solution are timely wiped off, so that the hypochlorous acid solution is prevented from being corroded and polluting the working surface for a long time;
S4, verifying surface purification effect
Immediately detecting the content of the nucleic acid remained on the working surface near the No. 1 air suction point and the No. 2 air suction point by adopting fluorescent quantitative PCR sampling after the surface purification treatment in the step S3; wherein, the sampling points of the working surfaces near the No. 1 air suction point and the No. 2 air suction point are 10, and the 10 sampling points are distributed on the following 3 working surfaces near each air suction point: ground (5 sampling points), stool (2 sampling points) and experiment operation table (3 sampling points); for the sampling point, a cotton swab wiping standard sampling method is adopted in sampling, namely, a surface with a sterile cotton swab wiping area of 5 cm multiplied by 5 cm is eluted in 1mL of sterile water, and fluorescent PCR detection is carried out on the eluent, wherein the detection result is as follows:
TABLE 9 detection results of nucleic acid residues on working surfaces in laboratory after surface purification
As shown in table 9, near the two air suction points, the detection results of 20 sampling points of 6 different working surfaces are all negative, and the surface purification in the surface step S3 is thorough, and then the three-dimensional purification effect verification can be performed in the step S5;
s5, three-dimensional purification effect verification
Immediately after the detection results of the step S4 are negative, 2 h, detecting the content of the nucleic acid remained on the working surfaces near the air suction point No. 1 and the air suction point No. 2 by adopting fluorescent quantitative PCR, wherein the number of the sampling points of the working surfaces near the air suction point No. 1 and the air suction point No. 2 is 20, and the 20 sampling points are distributed on the following 3 working surfaces near each air suction point: ground (12 sampling points), stool (4 sampling points) and experiment operation table (6 sampling points); for the sampling point, a cotton swab wiping standard sampling method is adopted in sampling, namely, a surface of a working surface with the area of 5 cm multiplied by 5 cm is wiped by a sterile cotton swab and eluted in 1mL of sterile water, and fluorescent PCR detection is carried out on eluent, wherein the detection result is as follows:
Table 10 test results of three-dimensional purification effect verification
As can be seen from Table 10, the detection results of 40 sampling points of 6 different working surfaces were all negative in the vicinity of two air suction points, indicating that there was no false negative purification result, and nucleic acid contamination in the laboratory enclosed space was thoroughly purified.
Claims (10)
1. The three-dimensional purification method for the nuclear acid pollution in the closed space of the laboratory is characterized by comprising the following steps:
s1, space purification: according to the size of the laboratory enclosed space, different numbers of air suction points are uniformly arranged, air in the laboratory enclosed space is sucked into the nucleic acid scavenger at the air suction points, the air is controlled to stay in the nucleic acid scavenger at least for 3 s, and the air is discharged into the laboratory after being contacted and mixed with the nucleic acid scavenger;
s2, verifying the space purifying effect: after the space purification in the step S1 is finished, detecting the content of residual nucleic acid in the air at each air suction point in the closed space respectively, and if the detection results are negative, entering the step S3; if the positive result is detected, repeating the step S1 and the step S2 until the detection results of all the air suction points in the step S2 are negative;
s3, surface purification: spraying the nucleic acid scavenger to a working surface in a laboratory closed space in a spray mode, so that the nucleic acid scavenger uniformly wets the working surface, and naturally airing the nucleic acid scavenger on the working surface;
S4, verifying the surface purification effect: immediately sampling and detecting the content of the residual nucleic acid of the working surface near each air suction point after the surface cleaning treatment in the step S3, wherein the number of the sampling points of the working surface near each air suction point is 10-15, and the 10-15 sampling points are distributed on at least 3 different working surfaces near the air suction point; if the detection results are negative, the step S5 is carried out; if the positive result is detected, repeating the step S3 and the step S4 until the detection result of the step S4 is negative;
s5, verifying the three-dimensional purification effect: immediately sampling and detecting the content of the residual nucleic acid of the working surface near each air suction point after the detection results in the step S4 are all negative and 2-4 h, wherein the number of sampling points of the working surface near each air suction point is 20-30, and the 20-30 sampling points are distributed on at least 3 different working surfaces near the air suction point; if all the detection results are negative, the three-dimensional purification of the acid pollution in the closed space is finished; if the positive result is detected, repeating the steps S1 to S5 until the detection results of the step S5 are all negative.
2. The method for three-dimensional purification of nucleic acid contamination in a confined laboratory space according to claim 1, wherein in step S1, the nucleic acid scavenger is a holomorphic acid solution or hypochlorous acid solution; in the step S3, the nucleic acid scavenger is a holohydronuclease solution or a hypochlorous acid solution.
3. The method for three-dimensional purification of acid contamination in a laboratory enclosed space according to claim 1, wherein in step S1, the air suction points synchronously suck air in the laboratory enclosed space at the same air suction rate.
4. The method for three-dimensional purification of acid contamination in a laboratory enclosed space according to claim 3, wherein in step S1, each 40 to 100 m of the laboratory enclosed space is obtained 3 An air suction point is provided.
5. A method for three-dimensional purification of acid contamination in a laboratory enclosed space according to claim 3, wherein in step S1, the total amount of air sucked is controlled to be 2 to 5 times that of the laboratory enclosed space.
6. The method for three-dimensional purification of nucleic acid contamination in a laboratory enclosed space according to claim 3, wherein in the step S1, the volume ratio of the nucleic acid scavenger to the laboratory enclosed space is 1 to 5:100L/m 3 。
7. The method for three-dimensional purification of acid contamination in a laboratory enclosed space according to claim 1, wherein the working surface is the outer surface of facilities, equipment, vessels and the outer surface of the clothing of an operator used for performing experimental operations in the laboratory enclosed space.
8. The method for three-dimensional purification of nucleic acid contamination in a laboratory enclosed space according to claim 1, wherein in step S4 and step S5, a high contamination area of the work surface in the vicinity of each air suction point is sampled by a cotton swab wiping standard sampling method to detect the content of nucleic acid remaining on the work surface.
9. The method for three-dimensional purification of acid pollution in a laboratory enclosed space according to claim 8, wherein the sampling area of the high pollution area of the working surface is 3-8 cm x 3-8 cm; the high pollution area of the working surface comprises the outer surfaces of an experiment operation table top, instruments and equipment, a test tube rack, a pipettor, a refrigerator, an ultra-clean workbench, a door handle, a stool, the ground and an illumination switch.
10. The method for the three-dimensional purification of nucleic acid contamination in a confined laboratory space according to any one of claims 1 to 9, wherein the nucleic acid content is detected by fluorescent quantitative PCR.
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