CN115228262A - Three-dimensional purification method for nucleic acid pollution in laboratory closed space - Google Patents
Three-dimensional purification method for nucleic acid pollution in laboratory closed space Download PDFInfo
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- 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
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- B01D—SEPARATION
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- B01D53/34—Chemical or biological purification of waste gases
- B01D53/346—Controlling the process
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- 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
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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
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- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F24—HEATING; RANGES; VENTILATING
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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
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- 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
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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
- F24F8/95—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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Abstract
The invention provides a three-dimensional purification method for nucleic acid pollution in a laboratory closed space, which comprises the following steps: s1, space purification: uniformly arranging different numbers of air suction points according to the size of the closed space of the laboratory, sucking the air in the closed space of the laboratory into the nucleic acid scavenger at the air suction points, controlling the air to be at least retained in the nucleic acid scavenger for 3s, and discharging the air into the laboratory after the air is contacted and mixed with the nucleic acid scavenger; s2, verifying the space purification effect; s3, surface purification; s4, verifying the surface purification effect; and S5, verifying the three-dimensional purification 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 experiment 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 forming modes in a closed space, one is that 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 the nucleic acid aerosol; the second is that the nucleic acid is directly attached to the operation surface in the closed space, such as an experiment operation table, the ground, the surface of instrument equipment and the like, i.e., the nucleic acid can directly pollute the operation surface.
Aerosols are colloidal dispersions of solid, liquid microparticles suspended in a gaseous medium, typically having a particle size of 1nm to 1 mm. Nucleic acid aerosol is generated along with the nucleic acid amplification process, and the nucleic acid aerosol pollution is generated in the processes of air and liquid surface friction, centrifugal machine centrifugation, violent shaking of a reaction tube, PCR (polymerase chain reaction) uncapping, repeated sample suction of a liquid transfer device, leakage of pollutants and the like. Once formed, nucleic acid aerosols are not only suspended in gaseous media for extended periods of time, but also partially settle on solid phase surfaces such as laboratory tables, instruments, consumables, floors, and the like, with the circulation of air. Therefore, in the closed space, the contamination of nucleic acid is dynamically changed, and it is difficult to ensure the nucleic acid in the closed space to be completely removed and purified by the conventional purification means.
As a scientific research and clinical examination place, a molecular laboratory has high PCR use frequency, a sample and an amplification target are frequently subjected to repeated treatment in multiple batches, nucleic acid pollution is continuously accumulated in an experimental area, and the pollution risk is continuously increased. With the continuous improvement of the amplification efficiency and sensitivity of technologies such as PCR and the like, the occurrence frequency of false positive in detection results is higher and higher due to the generated nucleic acid pollution. False positives mean that the results of the experiment are not authentic and directly contribute to the economic loss of the laboratory. More seriously, once nucleic acid contamination has developed, it can cause contamination of the entire PCR laboratory and can be difficult to clean out completely in a short time, requiring even shut down of the laboratory for a period of time if necessary.
Disclosure of Invention
The nucleic acid can be directly attached to the operation surface to cause nucleic acid pollution on the operation surface, meanwhile, the nucleic acid can pollute the air in the form of nucleic acid aerosol, in addition, the nucleic acid aerosol can naturally settle to the operation surface, and the nucleic acid aerosol or the nucleic acid pollution on the operation surface can also spread into the air to form nucleic acid aerosol pollution, so the nucleic acid pollution in the closed space is always a dynamic complex process; at present, the disinfection of a closed space in a laboratory usually has a better effect on bacteria, viruses and the like, and no systematic method for nucleic acid pollution can ensure that 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 closed laboratory space, which can ensure the three-dimensional purification effect of the nucleic acid pollution in the closed laboratory space so as to avoid the occurrence of false positive detection or experiment failure caused by the nucleic acid pollution in the laboratory.
The invention is realized by the following technical scheme.
The three-dimensional purification method for nucleic acid pollution in the closed space of the laboratory is characterized by comprising the following steps:
s1, space purification: uniformly arranging different numbers of air suction points according to the size of the closed space of the laboratory, sucking the air in the closed space of the laboratory into the nucleic acid scavenger at the air suction points, controlling the air to be at least retained for 3s in the nucleic acid scavenger, and discharging the air into the laboratory after the air is contacted and mixed with the nucleic acid scavenger;
in the step, air at different air suction points in the closed space of the laboratory is sucked into the nucleic acid scavenger, so that the air carrying with the pollution of the nucleic acid aerosol is fully contacted, mixed and reacted with the nucleic acid scavenger, and the nucleic acid is degraded, so that purified air can be discharged, and the effect of purifying the closed space of the laboratory is achieved;
s2, verifying the space purification effect: after the space purification in the step S1 is finished, respectively detecting the content of residual nucleic acid in the air of each air suction point in the closed space, if the detection results are negative, entering the step S3, and if positive results are detected, repeatedly circulating the step S1 and the step S2 until the detection results of all the air suction points in the step S2 are negative;
as the weight and the particle size of the nucleic acid aerosol in the air are possibly increased continuously after the nucleic acid aerosol is suspended in the air for a long time, part of the nucleic acid aerosol continuously and slowly settles to the working surface, thereby polluting the working surface; the sedimentation pollution of the nucleic acid aerosol seems to be uncontrollable, and the sedimentation pollution of the nucleic acid aerosol from a subsequent space to an operation surface can be avoided only after the nucleic acid aerosol in the air is thoroughly polluted and purified through the step S1 of space purification; otherwise, if the space is not thoroughly purified, the nucleic acid aerosol in the space still can be settled to pollute the surface after the surface is thoroughly purified in the step S3; therefore, after the space purification of step S1 is completed, the residual nucleic acid content in the air at each air suction point in the closed space needs to be detected to ensure the space purification effect of step S1;
s3, surface purification: spraying the nucleic acid scavenger to an operation surface in a closed laboratory space in a spraying manner, so that the operation surface is uniformly wetted by the nucleic acid scavenger, and naturally airing the nucleic acid scavenger on the operation surface;
in the experimental operation process, the nucleic acid may directly contaminate the working surface, or the nucleic acid aerosol suspended in the air settles and contaminates the working surface, which can cause the working surface in the closed space of the laboratory to be contaminated by the nucleic acid aerosol; by spraying the nucleic acid scavenger onto the working surface in the closed laboratory space in the form of a spray, the nucleic acid scavenger can be effectively brought into contact with the working surface while the amount of the nucleic acid scavenger used is reduced, and the nucleic acid aerosol settled on the working surface can be degraded;
s4, surface purification effect verification: immediately sampling and detecting the content of the nucleic acid remained on the working surface near each air suction point after the surface purification treatment in the step S3, wherein the number of 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 a positive result is detected, the steps S3 and S4 are repeatedly circulated until the detection results of the step S4 are negative;
after the surface purification is finished, immediate sampling is needed to verify whether the surface purification in the step S3 is thorough; if the surface purification in the step S3 is not complete, there is a possibility that the nucleic acid contamination of some working surfaces still exists, and in the subsequent detection or experiment operation process, along with the movement of personnel and inevitable touch on the working surfaces, there is still a certain risk that the nucleic acid contamination remaining on the working surfaces is resuspended in the air along with the flow of the air, so that the three-dimensional purification of the closed space of the laboratory cannot be realized, and therefore, the step S4 is required to verify whether the surface purification is complete;
s5, verifying the three-dimensional purification effect: immediately sampling and detecting the content of the nucleic acid remained on the working surface near each air suction point 2-4 hours after the detection results are negative in the verification of the surface purification effect in the step S4, 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 nucleic acid pollution in the closed space is finished; if a positive result is detected, repeating the steps S1 to S5 until the detection result of the step S5 is negative.
Since the steps S1 to S4 are sequentially completed, however, the nucleic acid aerosol remaining in the air inevitably settles and pollutes the working surface, and the remaining nucleic acid aerosol or nucleic acid pollution on the working surface risks to pollute the air with the movement of personnel or the touch with the working surface, there may be the following cases: when the surface purification is performed in step S3, since a person needs to perform necessary operations and touch the operation surface, the nucleic acid contamination on the operation surface will be resuspended in the air along with the flow of the air in the process of surface purification, and will not be easily deposited on the operation surface in a short time, so that the results of the surface purification effect verification in the subsequent step S4 are all negative, that is, although the results of the space purification effect verification in step S2 and the verification results of the surface purification effect verification in step S4 are both negative, it still cannot be ensured that the three-dimensional purification of nucleic acid contamination in the closed space of the laboratory is thorough, that is, there may be a case of false negative purification results; in contrast, the applicant samples and detects the content of the nucleic acid remained on the working surface near different suction points after the last cycle treatment of surface purification in the step S3 for 2-4 hours, if the air is polluted again during the surface purification in the step S4, the working surface is sampled after the detection results in the step S4 are negative for 2-4 hours, and the positive results are inevitably detected in the step S5 due to the sedimentation pollution of the nucleic acid aerosol; if the detection results in the step S5 are negative, the fact that the nucleic acid pollution in the closed space of the laboratory is three-dimensionally and thoroughly purified is shown;
in addition, the applicant has considered the following conditions and timings for verifying the three-dimensional purification effect in step S5: if a positive result is detected in the verification of the surface purification effect in the step S4, the surface purification in the closed space of the laboratory is not thorough, namely nucleic acid pollution of certain working faces still exists, and the step S5 is unnecessary in order to save unnecessary experimental links and purification time; and only when the detection results are negative in the verification of the surface purification effect in the step S4, the verification of whether the false negative purification result exists or not needs to be carried out in the step S5. In order to judge whether the air is polluted by the nucleic acid aerosol or not at the detection time, the nucleic acid aerosol is sampled by adopting a nucleic acid aerosol natural sedimentation method in the step S2, namely, a blank culture dish is placed in the space to be detected for a period of time, and the nucleic acid aerosol is naturally sedimented into the culture dish so as to be collected and further detected, the preferable range of sedimentation time (culture dish placement time) is discussed in the embodiment of the invention, and the result shows that the sedimentation time is 15-20 min, so that the pollution of the nucleic acid aerosol in the air can be detected; however, since the purpose of the verification in step S5 is to determine whether or not there is a false negative result, the false negative result may be a small amount of nucleic acid aerosol in the air because it is "nucleic acid contamination on the work surface is resuspended in the air with the flow of the air during the surface decontamination and is difficult to completely settle to the work surface in a short time", and therefore if the detection is performed by collecting nucleic acid aerosol by natural settling as well, the settling time should be increased so that the nucleic acid aerosol diffused to the air has a sufficient settling time to settle to the work surface. In contrast, the applicant comprehensively considers the factors and selects to sample the operation surface 2-4 hours after the detection results in the step S4 are all negative, namely, at the moment, if nucleic acid aerosol pollution exists in the air, the nucleic acid aerosol has settled for 2-4 hours (which is far more than 15-20 min), so that effective nucleic acid pollution can be collected from the operation surface; in addition, the applicant also increases the number of sampling points in step S5, and controls the number of sampling points on 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 totipotent nuclease solution (UNC enzyme solution) or a hypochlorous acid solution; in the step S3, the nucleic acid scavenger is a totipotent nuclease solution (UNC enzyme solution) or a hypochlorous acid solution.
Preferably, in step S2, the number of sampling points of each air suction point is 10 to 15, and the 10 to 15 sampling points should be distributed at least 3 different spatial positions near the air suction point.
Preferably, in step S2, sampling modes of the sampling points are as follows: a blank culture dish is placed at a sampling point, nucleic acid aerosol in the air naturally settles for 15-20 min to the blank culture dish, and then whether nucleic acid pollution exists in the air is detected and represented by whether nucleic acid pollution exists in the culture dish.
Preferably, in step S2, the number of sampling points of each air suction point is 10 to 15, and the 10 to 15 sampling points should be distributed at least 3 different spatial positions near the air suction point.
Preferably, in the step S1, the air suction points synchronously suck the air in the laboratory enclosed 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, thereby improving the efficiency of space purification.
Preferably, in the step S1, the volume of the laboratory enclosed space is 40 to 100m 3 An air suction point is provided.
Preferably, in the step S1, the total suction amount of the air is controlled to be 2 to 5 times of the laboratory closed space.
Preferably, in the step S1, the ratio of the volume of the nucleic acid scavenger used to the volume of the laboratory enclosed space is 1 to 5 3 。
Preferably, in step S2, the working surface is an outer surface of a facility, equipment, or vessel used for performing an experimental operation in a laboratory enclosed space, or an outer surface of a garment of an operator.
Preferably, in the steps S4 and S5, a cotton swab wiping standard sampling method is used to sample the high-pollution area of the working surface so as to detect the content of the remaining nucleic acid on the working surface.
Preferentially, in the step S2, 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 working face includes experiment operation mesa, instrument and equipment, test-tube rack, pipettor cell-phone shell, refrigerator, superclean bench, door handle, stool, ground and lighting switch's surface.
Preferably, the nucleic acid content is detected by fluorescent quantitative PCR.
The invention has the following beneficial effects:
1) According to the invention, nucleic acid pollution in air in a closed laboratory space can be effectively purified through space purification, and nucleic acid pollution on a working surface in the laboratory can be effectively purified through surface purification, so that three-dimensional purification of the laboratory space and the surface is realized.
2) The risk that the nucleic acid aerosol in the air pollutes the working surface again is higher because the nucleic acid aerosol in the air is inevitably settled and polluted; and the surface diffusion pollution only occurs with a certain probability, and the risk of the diffusion pollution of the air is lower. Therefore, aiming at the characteristics that the nucleic acid aerosol has high-risk air sedimentation pollution and low-risk surface diffusion pollution, the invention firstly eliminates the high-risk nucleic acid aerosol sedimentation pollution through space purification, carries out effect verification, then carries out surface purification to eliminate the low-risk nucleic acid diffusion pollution, simultaneously carries out effect verification, can ensure thorough three-dimensional purification of nucleic acid pollution in a closed space of a laboratory through scientific control of the purification and verification steps, and finally carries out three-dimensional purification effect verification to avoid the occurrence of false positive detection or experimental failure caused by nucleic acid pollution in the laboratory.
Drawings
FIG. 1 is a flow chart of the method of the present invention.
Detailed Description
The technical solutions in the present invention are described clearly and completely below, and it is obvious that the described embodiments are only some of the embodiments of the present invention, not all of them. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
Referring to fig. 1, the method for three-dimensional purification of nucleic acid pollution in a laboratory enclosed space comprises the following steps:
s1, space purification
Uniformly arranging different numbers of air suction points according to the size of the closed space of the laboratory, sucking the air in the closed space of the laboratory into the nucleic acid scavenger at the air suction points, controlling the air to be at least retained in the nucleic acid scavenger for 3s, and discharging the air into the laboratory after the air is contacted and mixed with the nucleic acid scavenger;
s2, verifying space purification effect
After the space purification in the step S1 is finished, respectively detecting the content of residual nucleic acid in the air of each air suction point in the closed space, if the detection results are negative, entering the step S3, and if positive results are detected, repeatedly circulating 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 an operation surface in a closed laboratory space in a spraying manner, so that the operation surface is uniformly wetted by the nucleic acid scavenger, and naturally airing the nucleic acid scavenger on the operation surface;
s4, verifying surface purification effect
Immediately sampling and detecting the content of the nucleic acid remained on the working surface near each air suction point after the surface purification treatment in the step S3, wherein the number of 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 a positive result is detected, the steps S3 and S4 are repeatedly circulated until the detection results of the step S4 are negative;
s5, verifying three-dimensional purification effect
Immediately sampling and detecting the content of the nucleic acid remained on the working surface near each air suction point 2-4 hours after the detection results are all negative in the step S4, wherein the number of the 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 nucleic acid pollution in the closed space is finished; if a positive result is detected, repeating the steps S1 to S5 until the detection result of the step S5 is negative.
Example 2
Referring to fig. 1, the method for three-dimensional purification of nucleic acid pollution in a laboratory enclosed space comprises the following steps:
s1, space purification
Uniformly arranging different numbers of air suction points according to the size of the closed space of the laboratory, sucking the air in the closed space of the laboratory into the nucleic acid scavenger at the air suction points, controlling the air to be at least retained for 3s in the nucleic acid scavenger, and discharging the air into the laboratory after the air is contacted and mixed with the nucleic acid scavenger; wherein, different numbers of air suction points are uniformly arranged according to the size of the closed space of the laboratory, namely every 40 to 100m in the closed space of the laboratory 3 Setting an air suction point, synchronously sucking air in the closed laboratory space at different air suction points at the same air suction rate, and controlling the total suction amount of the air to be the closed laboratory space2 to 5 times, the ratio of the volume of the nucleic acid scavenger used to the volume of the laboratory enclosed space is 1 to 5 3 (ii) a The nucleic acid scavenger is UNC enzyme solution or hypochlorous acid solution; when UNC enzyme solution is used as the nucleic acid scavenger, the concentration of the UNC enzyme is preferably 3-4 ug/mL; when using hypochlorous acid solution as nucleic acid scavenger, the recommended concentration of hypochlorous acid solution is 400-600 ppm;
s2, verifying space purification effect
After the space purification in the step S1 is finished, respectively detecting the content of residual nucleic acid in the air of each air suction point in the closed space by adopting fluorescence quantitative PCR, if the detection result is negative, entering the step S3, and if a positive result is detected, repeatedly circulating the step S1 and the step S2 until the detection result in the step S2 is 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 to the blank culture dish, and further representing whether the air is polluted by nucleic acid according to the content of the nucleic acid in the culture dish;
s3, surface purification
If the UNC enzyme solution is used as a nucleic acid scavenger, and the concentration of the UNC enzyme is preferably 3-4 ug/mL, spraying the UNC enzyme solution to an operation surface in a closed laboratory space in a spraying manner, so that the UNC enzyme solution uniformly wets the operation surface, forming a water film on the operation surface, and naturally airing the UNC enzyme solution on the operation 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 operation surface in a laboratory closed space in a spraying mode, so that the hypochlorous acid solution uniformly wets the operation surface, after the hypochlorous acid solution is naturally dried on the operation surface, the operation surface needs to be wiped by pure water, residues of the hypochlorous acid solution are wiped off, and therefore the long-time residual corrosion and the pollution to the operation surface of the hypochlorous acid solution are prevented;
the operation surface is the outer surfaces of facilities, equipment and vessels used for experimental operation in a laboratory closed space and the outer surfaces of clothes of operators;
s4, verifying surface purification effect
Immediately sampling and detecting the content of the nucleic acid remained on the working surface near each air suction point after the surface purification treatment in the step S3, wherein the number of 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 a positive result is detected, the steps S3 and S4 are repeatedly circulated until the detection results of the step S4 are negative; sampling a high-pollution area of a working surface near each air suction point by adopting a cotton swab wiping standard sampling method to detect the content of nucleic acid remained on 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 an experimental operation table surface, instrument equipment, a test tube rack, a pipettor, a mobile phone shell, a refrigerator, an ultra-clean workbench, a door handle, a stool, a ground and the outer surface of a lighting switch;
s5, verifying three-dimensional purification effect
Immediately sampling and detecting the content of the nucleic acid remained on the working surface near each air suction point 2-4 hours after the detection results are all negative in the step S4, wherein the number of the 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 nucleic acid pollution in the closed space is finished; if a positive result is detected, repeating the steps S1 to S5 until the detection result of the step S5 is negative; the sampling area of the high-pollution area of the operation surface is 3-8 cm multiplied by 3-8 cm, and the high-pollution area of the operation surface comprises an experiment operation table surface, instrument equipment, a test tube rack, a pipettor, a mobile phone shell, a refrigerator, an ultra-clean workbench, a door handle, a stool, a ground and the outer surface of an illumination switch.
Example 3
Influence of different sedimentation time on detection results of nucleic acid aerosol natural sedimentation sampling.
1. The principle of the natural sedimentation method of nucleic acid aerosol: by utilizing the characteristic of continuous natural sedimentation of nucleic acid aerosol in the air, the nucleic acid sedimented to the culture dish from the air is collected by the blank culture dish which is placed in the air for a period of time (sedimentation time), and then whether nucleic acid aerosol pollution exists in the air is represented by detecting whether the nucleic acid exists in the culture dish, if the nucleic acid exists in the blank culture dish, the nucleic acid pollution exists in the air is represented, otherwise, the nucleic acid pollution does not exist in the air.
2. The detection method comprises the following steps: 300mL of PCR fragment test solution with a Ct value of 20.6 was prepared by using a CN61 aerosol generator and all the PCR fragment test solutions had a space size of 79m 3 Spraying treatment is carried out in the left and right closed laboratories, then 9 blank culture dishes (the specification of the culture dishes is 150mm, the culture dishes are strictly disinfected and have no nucleic acid pollution) are randomly and simultaneously placed in the closed laboratories, after the culture dishes are sequentially placed for 15min, 20min and 30min, any 3 culture dishes are respectively taken out from the 9 culture dishes, samples are immediately collected by cotton swabs and eluted in 1mL of sterile water, the eluent is taken for carrying out fluorescence quantitative PCR detection to discuss the change of the detected nucleic acid content in the culture dishes under different settling times, and the results are shown in Table 1; in the present invention, the manufacturer of the CN61 aerosol generator is Collison corporation of usa; the model of the real-time fluorescent quantitative PCR instrument is a full-automatic medical PCR analysis system SLAN-96S;
TABLE 1 results of detection of nucleic acid residues in air
As can be seen from Table 1, after the nucleic acid pollution source is sprayed to the closed space, the nucleic acid naturally settles, and the settled nucleic acid is collected by using a culture dish, and the detection results in different settling times show that the longer the settling time is, the smaller the Ct value is, the higher the detected nucleic acid content is. Although the Ct value is reduced in 15min of sedimentation compared with 20min of sedimentation, the reduction range is small, the result of 20min of sedimentation is not significantly different from that of 30min of sedimentation, but nucleic acid can be detected in a culture dish in 15 min-30 min of sedimentation, so that in order to save detection time and ensure that nucleic acid pollution in the air can be detected, 15 min-20 min can be selected as sedimentation time length to detect nucleic acid pollution in the air.
Example 4
The effect of space purification in the method is verified.
1. Modeling of nucleic acid-contaminated spaces
Selecting two spaces with the size of 79m 3 Two 300mL bottles of PCR fragment test solution with Ct value of about 20 are respectively sprayed into the closed space of two laboratories and the surface of equipment facilities by a CN61 aerosol generator in a spraying mode by taking the left and right closed laboratories as processing objects, wherein one of the two laboratories is used as a comparison group, and the other one is used as an experiment group;
2. spatial decontamination of nucleic acids
The experimental group adopts the step S1 in the method of the invention to carry out space purification, while the comparative group does not adopt any measures; specifically, the space purification method comprises the following steps:
first, 3L of a nucleic acid scavenger (the nucleic acid scavenger is a UNC enzyme solution having a UNC enzyme concentration of 3.4ug/mL, and 25mm Tris-HCl (pH = 8.2), 10mm NaCl, and 2mm MgCl 2 Buffer solution at concentration) is placed in a container selected to have a narrow mouth and a deep bottom to ensure adequate contact time of the aspirated air with the nucleic acid scavenger; the volume of the laboratory closed space of the experimental group is 79m 3 The left and the right, therefore, only one air suction point is needed to be arranged, and the air suction point is positioned at the middle position of the laboratory closed space of the experimental group as much as possible; pumping air into the bottom of the container at the air suction point by using a pump body, and allowing the pumped air to be retained in the nucleic acid scavenger for at least 3s and then automatically discharged; in order to ensure that the polluted air in the laboratory closed space of the laboratory group can be pumped into the container as much as possible, the total air suction amount is controlled to be 3 times of the laboratory closed space, and only reasonable control is needed under the condition that the air suction efficiency of the pump body and the volume of the laboratory closed space of the laboratory group are determinedThe air exhaust time is only needed, and the space purification time set by the embodiment is 60min.
3. Determination of results
After the space purification of the experimental group is finished, the content of the residual nucleic acid in the air of 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 contrast group is detected by adopting fluorescent quantitative PCR, and the specific process is as follows:
for the experimental group, after the space purification of the experimental group is completed, 10 blank culture dishes (the specification of the culture dishes is 150mm, the culture dishes are strictly disinfected and have no nucleic acid pollution) are randomly placed on the experimental operation platform near the suction point and the ground, so that the sampling points are distributed at different space positions of the air suction point, after the culture dishes are kept still for 20min, the culture dishes are taken out, samples are collected by cotton swabs and eluted in 1mL sterile water, the eluent is taken for fluorescence quantitative PCR detection, and the result is shown in Table 2;
for the comparison group, the same sampling method, sampling time and detection mode as those of the experimental group were adopted, and the results are shown in table 2;
TABLE 2 detection of the effectiveness of spatial decontamination
As can be seen from Table 2, PCR fragments could not be detected in the blank culture dish after the space purification of the experimental group, so that the nucleic acid pollution in the air in the laboratory closed space of the experimental group can be thoroughly purified by the space purification in the invention. Generally, in the present invention, the nucleic acid contamination in the air in the closed laboratory space can be effectively purified by the step S1 space purification, but it is necessary to perform the standard operation by the experimenter under the set parameter conditions, so that it is inevitable that the space purification is not complete due to human factors or unexpected objective factors, and therefore, the step S2 space purification effect verification is required after the step S1 space purification is performed to determine whether the nucleic acid in the air is completely purified.
Example 5
The effect of surface purification in the method of the invention is verified.
1. Modeling of nucleic acid-contaminated surfaces
On different working surfaces in a laboratory, including the outer surfaces of a ground, a refrigerator, a door handle, an ep pipe frame and an experiment operation table, 100uL of PCR fragment test solution (Ct value is about 20) is sprayed on the different working surfaces in an area of 5cm multiplied by 5cm, after the PCR fragment test solution is naturally aired, a sterile cotton swab is used for wiping the whole surface, sampling is carried out in 1mL of sterile water for elution, the eluent is subjected to fluorescence PCR detection, the nucleic acid pollution condition of the surface before purification is determined, and the result is shown in a table 2;
2. surface decontamination of nucleic acids
Adopting hypochlorous acid solution as a nucleic acid scavenger, wherein the using concentration of the hypochlorous acid solution is 500ppm, according to the method of the step S3, spraying 0.4mL of hypochlorous acid solution to the area of 5cm multiplied by 5cm of the different operation surfaces in a spraying mode, enabling the hypochlorous acid solution to uniformly wet the operation surfaces, naturally airing the hypochlorous acid solution on the operation surfaces (the airing time after spraying is controlled to be 30 min), wiping the whole surface with a sterile cotton swab, sampling the whole surface, eluting the whole surface in 1mL of sterile water, carrying out fluorescence PCR detection on the eluent, and measuring the nucleic acid pollution condition of the purified surface, wherein the result is shown in Table 3;
3. determination of results
TABLE 3 examination of the effectiveness of surface decontamination
| Sampling point | Ct value before purification | Ct value after purification |
| Ground surface | 29.07 | NoCt |
| Refrigerator | 28.10 | NoCt |
| Door handle | 30.22 | NoCt |
| ep pipe frame | 31.18 | NoCt |
| Stool | 31.59 | NoCt |
| Experiment operating table | 28.53 | NoCt |
As can be seen from the results in Table 3, the sampling test results of the working surface are negative after the surface is purified by the method of the present invention, which indicates that the method has a good effect of removing the nucleic acid contamination on the solid surface. However, in the present invention, the surface purification in step S3 needs to be completed by experimenters through standard operations under the set parameter conditions, so there are inevitable situations where the spatial purification is not complete due to artificial reasons or unexpected objective factors, such as uneven spraying on the working surface, or some working surfaces are not sprayed, and therefore, after the surface purification in step S3, verification of the surface purification effect in step S4 needs to be performed to determine whether the nucleic acid contamination on the working surface is complete.
Example 6
Selecting a space with the size of 79m 3 The left and right laboratories are used as processing objects300mL of PCR fragment test solution with a Ct value of about 20 is sprayed into a closed space of a laboratory and on a working surface in a spraying mode by a CN61 aerosol generator, so that the laboratory forms nucleic acid pollution, and then the laboratory of an experimental group is subjected to three-dimensional purification by adopting the method disclosed by the invention, please refer to figure 1, wherein the specific process is as follows:
s1, space purification
The nucleic acid scavenger was UNC enzyme solution with a UNC enzyme concentration of 3.4ug/mL using 25mm Tris-HCl (pH = 8.2), 10mm NaCl,2mm MgCl 2 Preparing a buffer solution with a concentration; the 3L of nucleic acid scavenger is first placed in a container, the container being selected to have a narrow mouth and a deep bottom to ensure adequate contact time between the drawn air and the nucleic acid scavenger; the volume of the laboratory closed space of the experimental group is 79m 3 The left and the right, therefore, only one air suction point is needed to be arranged, and the air suction point is positioned at the middle position of the laboratory closed space of the experimental group as much as possible; pumping air into the bottom of the container at the air suction point by using a pump body, and allowing the pumped air to be retained in the nucleic acid scavenger for at least 3s and then automatically discharged; in the process of pumping air to discharge, the nucleic acid aerosol carried in the air can contact, mix and react with the nucleic acid scavenger, so that the nucleic acid in the air is degraded, and finally purified air can be discharged, thereby achieving the effect of purifying the closed space of a laboratory group; in order to ensure that the polluted air in the laboratory closed space of the laboratory group can be pumped into the container as far as possible, the total air suction quantity is controlled to be 2-5 times of the laboratory closed space, the air suction time only needs to be reasonably controlled under the condition that the air suction efficiency of the pump body and the volume of the laboratory closed space of the laboratory group are determined, and the space purification time is set to be 60min in the embodiment.
S2, verifying space purification effect
After the space purification in the step S1 is finished, detecting the content of the residual nucleic acid in the air of the air suction point by adopting fluorescence quantitative PCR, wherein the specific process is as follows:
after the spatial purification in the step S1 is finished, 10 blank culture dishes (the specification of the culture dishes is 150mm, the culture dishes are strictly disinfected and have no nucleic acid pollution) are immediately placed on an experiment operation table, the ground and stools near the suction point, 4 blank culture dishes are placed on a table among the 10 blank culture dishes, the number of the blank culture dishes is 1-4, 4 blank culture dishes are placed on the ground, the number of the blank culture dishes is 5-8, 2 blank culture dishes are placed on the stools, the number of the blank culture dishes is 9-10, sampling points are distributed at different spatial positions near the air suction point, after the culture dishes are kept still for 20min, the culture dishes are taken out and eluted in 1mL of sterile water by using cotton swabs, and the eluents are taken for fluorescence quantitative PCR detection, wherein the results are shown in a table 4;
TABLE 4 detection results of nucleic acid residues in laboratory air after space purification
As can be seen from Table 4, after the space purification in step S1, PCR fragments 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 has been completely completed in step S1, and the process can proceed to step S3.
S3, surface purification
The nucleic acid scavenger was UNC enzyme solution with a UNC enzyme concentration of 3.4ug/mL using 25mm Tris-HCl (pH = 8.2), 10mm NaCl,2mm MgCl 2 Preparing a buffer solution with a concentration; spraying the nucleic acid scavenger to an operation surface in a laboratory closed space in a spraying mode to enable the nucleic acid scavenger to uniformly wet the operation surface and form a water film on the operation surface, and naturally airing the nucleic acid scavenger on the operation surface; the operation surface is the outer surfaces of facilities, equipment and vessels used for experimental operation in a laboratory closed space and the outer surfaces of clothes of operators;
s4, verifying surface purification effect
Immediately after the surface purification treatment in step S3, detecting the content of the nucleic acid remaining on the working surface near the air suction point by using fluorescence quantitative PCR sampling, wherein the number of the sampling points on the working surface near the air suction point is 10, and the 10 sampling points are distributed on the following 6 working surfaces near the air suction point: the method comprises the following steps of (1) carrying out on-line detection on the ground (3 sampling points), a refrigerator (2 sampling points), a door handle (1 sampling point), an ep pipe frame (1 sampling point), a stool (1 sampling point) and an experimental operating platform (2 sampling points); for the sampling points, a cotton swab wiping standard sampling method is adopted during sampling, namely, a sterile cotton swab is used for wiping the surface with the area of 5cm multiplied by 5cm and is eluted in 1mL of sterile water, the eluent is subjected to fluorescence PCR detection, and the detection result is as follows:
TABLE 5 results of detection of nucleic acid residues on laboratory work surface after surface decontamination
| Serial number | Sampling point | Ct value of Experimental group |
| 1 | Floor 1 | NoCt |
| 2 | Floor 2 | NoCt |
| 3 | Floor 3 | NoCt |
| 4 | Refrigerator 1 | NoCt |
| 5 | Refrigerator 2 | NoCt |
| 6 | Door handle | NoCt |
| 7 | ep pipe frame | NoCt |
| 8 | Stool | NoCt |
| 9 | Experiment operating table 1 | NoCt |
| 10 | Experiment operating table 2 | NoCt |
As can be seen from table 5, if the detection results of 10 sampling points of 6 different working surfaces are negative, the step S5 is performed to verify the three-dimensional purification effect;
s5, verifying three-dimensional purification effect
Immediately sampling 2h after the detection results in the step S4 are all negative, and detecting the content of the nucleic acid remained on the working surface near the air suction point by adopting fluorescence quantitative PCR, wherein the number of the sampling points of 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: the method comprises the following steps of (1) carrying out on-line detection on the ground (8 sampling points), the refrigerator (2 sampling points), the door handle (1 sampling point), the ep pipe frame (1 sampling point), the stool (2 sampling points) and the experiment operating table (6 sampling points); for the sampling points, a cotton swab wiping standard sampling method is adopted during sampling, namely, a sterile cotton swab is used for wiping the surface of an operation surface with the area of 5cm multiplied by 5cm and is eluted in 1mL of sterile water, the eluent is subjected to fluorescence PCR detection, and the detection result is as follows:
table 6 test results of three-dimensional purification effect verification
As can be seen from Table 6, the detection results of 20 sampling points of 6 different working planes are all negative, which indicates that no false negative purification result exists and the nucleic acid pollution in the closed space of the laboratory is completely purified.
Example 7
The size of the selected space is 150m 3 The laboratory of the left, right, rectangle house type is regarded as the processing object, 800mL PCR fragment test solution with Ct value about 20 prepared is sprayed to the closed space and working face of the laboratory with CN61 aerosol generator in the form of spray, thus make the laboratory form the nucleic acid pollution, then use the method of the invention to carry on the stereoscopic purification to the laboratory of the experimental group, please refer to figure 1, the concrete process is as follows:
s1, space purification
The nucleic acid scavenger was UNC enzyme solution with a UNC enzyme concentration of 3.4ug/mL using 25mm Tris-HCl (pH = 8.2), 10mm NaCl,2mm MgCl 2 Preparing a buffer solution with a concentration; dividing 8L of the nucleic acid scavenger into two portions, and placing the portions in two separate containers, the containers being selected to have a narrow mouth and a deep bottom to ensure sufficient contact time between the drawn air and the nucleic acid scavenger; the volume of the laboratory closed space of the experimental group is 150m 3 On the left and right sides, only two air suction points, namely an air suction point No. 1 and an air suction point No. 2 are arranged, and the two air suction points are respectively positioned at the middle positions of the left side and the right side of the laboratory closed space of the experimental group; then pumping air into the bottom of the container containing 4L of nucleic acid scavenger at the two air suction points by using two pump bodies, respectively, so that the pumped air is retained in the nucleic acid scavenger for at least 3s, and thenAutomatically discharging; in the process of pumping air to discharge, the nucleic acid aerosol carried in the air can contact, mix and react with the nucleic acid scavenger, so that the nucleic acid in the air is degraded, and finally purified air can be discharged, thereby achieving the effect of purifying the closed space of a laboratory group; in order to ensure that the polluted air in the laboratory closed space of the laboratory group can be pumped into the container as far as possible, the total air suction quantity is controlled to be 2-5 times of the laboratory closed space, the air suction time only needs to be reasonably controlled under the condition that the air suction efficiency of the pump body and the volume of the laboratory closed space of the laboratory group are determined, and the space purification time is set to be 60min in the embodiment.
S2, verifying space purification effect
After the space purification in the step S1 is finished, detecting the content of the residual nucleic acid in the air of the air suction point by adopting fluorescence quantitative PCR, wherein the specific process is as follows:
after the space purification in the step S1 is finished, 10 blank culture dishes (the specification of the culture dishes is 150mm, and the culture dishes are strictly disinfected without nucleic acid pollution) are respectively placed on an experimental operating platform, 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 spatial positions near the air suction points; wherein 4 blank culture dishes with the number of 1-4 are placed on a table in 10 blank culture dishes with the number of 1-4; 4 pieces of the materials are placed on the ground, and the number of the materials is 5 to 8; 2 stools are arranged on the stool, and the number of the stools is 9-10; 4 blank culture dishes with the same number of No. 2 air suction points are placed on a table, and the numbers are 11-14; 4 pieces of the materials are placed on the ground, and the number of the materials is 15 to 18; 2 stools are arranged on the stool, and the number of the stools is 19-20; standing the blank culture dish for 20min, and taking out the culture dish; then, the blank culture dish taken out is eluted in 1mL of sterile water by using a cotton swab collection sample, and the eluent is taken for fluorescence quantitative PCR detection, wherein the results are shown in Table 7;
TABLE 7 detection results of nucleic acid residues in laboratory air after space purification
As is clear from Table 7, since PCR fragments were detected at the air suction point 2 after the space purging for 60min and 120min, i.e., positive detection was observed, the nucleic acids in the air in the closed space in the laboratory were not completely purged in step S1, and therefore, the space purging was performed by repeating step S1 again. The method for verifying the space 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 space purification
As can be seen from Table 8, after repeating step S1, i.e., after performing the second spatial purification, PCR fragments could not be detected in the blank petri dish, and thus the purification of nucleic acid contamination in air in the closed space of the laboratory of the experimental group was completely completed, and the process may proceed to step S3.
S3, surface purification
Using hypochlorous acid solution as a nucleic acid scavenger, wherein the recommended use concentration of the hypochlorous acid solution is 500ppm, spraying the hypochlorous acid solution to a working surface in a laboratory closed space in a spraying mode to enable the hypochlorous acid solution to uniformly wet the working surface, wiping the working surface with pure water after the hypochlorous acid solution is naturally dried on the working surface, and wiping off residues of the hypochlorous acid solution in time, so that the hypochlorous acid solution is prevented from remaining for a long time to corrode and pollute the working surface;
s4, verifying surface purification effect
Immediately adopting fluorescent quantitative PCR sampling to detect the content of nucleic acid remained on the working surface near the No. 1 air suction point and the No. 2 air suction point after the surface is purified in the step S3; the number of sampling points of the working surfaces near the No. 1 air suction point and the No. 2 air suction point is 10, and the 10 sampling points are distributed on the following 3 working surfaces near each air suction point: the system comprises a ground (5 sampling points), stools (2 sampling points) and an experimental operating platform (3 sampling points); for the sampling points, a cotton swab wiping standard sampling method is adopted during sampling, namely, a sterile cotton swab is used for wiping the surface with the area of 5cm multiplied by 5cm and is eluted in 1mL of sterile water, the eluent is subjected to fluorescence PCR detection, and the detection result is as follows:
TABLE 9 results of detection of nucleic acid residues on laboratory work surface after surface decontamination
As can be seen from table 9, in the vicinity of the two air suction points, the detection results of 20 sampling points of 6 different working surfaces are negative, and if the surface purification of step S3 is complete, step S5 can be performed to verify the three-dimensional purification effect;
s5, verifying three-dimensional purification effect
Immediately sampling 2h after the detection results of the step S4 are all negative, and detecting the content of the nucleic acid remained on the working surfaces near the No. 1 air suction point and the No. 2 air suction point by adopting fluorescence quantitative PCR, wherein the number of the sampling points of the working surfaces near the No. 1 air suction point and the No. 2 air suction point is respectively 20, and the 20 sampling points are distributed on the following 3 working surfaces near the air suction points: the system comprises a ground (12 sampling points), stools (4 sampling points) and an experimental operating platform (6 sampling points); for the sampling points, a cotton swab wiping standard sampling method is adopted during sampling, namely, a sterile cotton swab is used for wiping the surface of an operation surface with the area of 5cm multiplied by 5cm and is eluted in 1mL of sterile water, the eluent is subjected to fluorescence PCR detection, and 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 planes are all negative near two air suction points, which indicates that no false negative purification results exist and the nucleic acid pollution in the closed laboratory space is completely purified.
Claims (10)
1. The three-dimensional purification method for nucleic acid pollution in the closed space of the laboratory is characterized by comprising the following steps:
s1, space purification: uniformly arranging different numbers of air suction points according to the size of the closed space of the laboratory, sucking the air in the closed space of the laboratory into the nucleic acid scavenger at the air suction points, controlling the air to be at least retained in the nucleic acid scavenger for 3s, and discharging the air into the laboratory after the air is contacted and mixed with the nucleic acid scavenger;
s2, verifying the space purification effect: after the space purification in the step S1 is finished, respectively detecting the content of residual nucleic acid in the air of each air suction point in the closed space, and if the detection results are negative, entering a step S3; if a 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 an operation surface in a closed laboratory space in a spraying manner, so that the operation surface is uniformly wetted by the nucleic acid scavenger, and naturally airing the nucleic acid scavenger on the operation surface;
s4, surface purification effect verification: immediately sampling and detecting the content of the nucleic acid remained on the working surface near each air suction point after the surface purification treatment in the step S3, wherein the number of 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 a 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 residual nucleic acid content of the working surface near each air suction point 2-4 h after the detection results are negative in the step S4, 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 nucleic acid pollution in the closed space is finished; if a positive result is detected, repeating the steps S1 to S5 until the detection result of the step S5 is negative.
2. The method for three-dimensional purification of nucleic acid contamination in a laboratory enclosed space according to claim 1, wherein in step S1, the nucleic acid scavenger is a nuclease omnipotent solution or a hypochlorous acid solution; in the step S3, the nucleic acid scavenger is a totipotent nuclease solution or a hypochlorous acid solution.
3. The method for the three-dimensional purification of the nucleic acid contamination in the closed laboratory space according to claim 1, wherein in the step S1, the air suction points synchronously suck the air in the closed laboratory space at the same air suction rate.
4. The method for three-dimensional purification of nucleic acid contamination in the closed laboratory space according to claim 3, wherein in step S1, every 40-100 m of the closed laboratory space is used 3 An air suction point is provided.
5. 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 total suction amount of air is controlled to be 2 to 5 times of the laboratory enclosed space.
6. The method for the three-dimensional purification of nucleic acid contamination in a laboratory enclosed space according to claim 3, wherein in the step S1, the ratio of the volume of the nucleic acid scavenger used to the volume of the laboratory enclosed space is 1 to 5 3 。
7. The method for three-dimensional purification of nucleic acid contamination in a laboratory enclosed space according to claim 1, wherein in step S2, the working surfaces are the outer surfaces of facilities, equipment, vessels and the outer surfaces of clothing of operators used for performing experimental operations in the laboratory enclosed space.
8. The method for three-dimensional purification of nucleic acid contamination in the closed laboratory space according to claim 1, wherein in the steps S4 and S5, a cotton swab wiping standard sampling method is used to sample the high contamination area of the working surface near each air suction point so as to detect the content of the nucleic acid remaining on the working surface.
9. The method for three-dimensional purification of nucleic acid contamination in a laboratory enclosed space according to claim 9, wherein in step S2, the sampling area of the high contamination area of the work surface is 3 to 8cm x 3 to 8cm; the high-pollution area of the operation surface comprises an experiment operation table surface, instrument equipment, a test tube rack, a pipettor, a refrigerator, an ultra-clean workbench, a door handle, a stool, the ground and the outer surface of a lighting switch.
10. The method for the three-dimensional purification of nucleic acid contamination in a laboratory enclosed 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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