CN102634449B - Viral aerosol collecting and enriching instrument - Google Patents

Abstract

A viral aerosol collection and enrichment instrument includes a frame, a gas supply device, a gas outlet unit nozzle and a collection unit tube; the gas supply device is used to collect a gas sample to be detected, and the gas sample is input to the In the nozzle of the gas outlet unit; the nozzle of the gas outlet unit and the pipe of the collection unit are both arranged on the frame, and the pipe of the collection unit is detachably arranged under the nozzle of the gas outlet unit, and a test device with virus-capturing immune magnetic beads is arranged therein. liquid; the gas sample is sprayed into the collection unit tube in a rotating manner through the nozzle of the gas outlet unit, the test liquid absorbs the aerosol contained in the gas to be tested, and captures the gas to be tested through the virus immune magnetic beads virus carried. While efficiently capturing biological aerosols in the air, the invention can specifically realize the enrichment of virus and microorganism samples, and is especially suitable for the monitoring of virus pathogens in ambient air.

Description

Viral aerosol collection and enrichment instrument

technical field

The present invention relates to a viral aerosol collection and enrichment device, in particular to an integrated device that effectively collects biological aerosols in ambient air and integrates virus enrichment technology for subsequent virus detection.

Background technique

At present, most of the epidemics of infectious diseases are mainly caused by virus infection, and the virus infection with the respiratory tract as the main transmission route is very easy to spread rapidly in a short period of time due to its convenient mode of transmission and infection, leading to serious public health incidents. In recent years, SARS The serious health hazards and international concerns caused by the infection and spread of highly pathogenic avian influenza and the new type A H1N1 influenza virus are typical examples; the spread of virus-like bioterrorism factors through aerosol is also the focus of anti-terrorism experts. Guard content. In addition, non-respiratory viruses can also be infected through aerosols or caused by laboratory misuse and transmission from time to time. Therefore, it is urgent and necessary to study the virus monitoring of aerosols in the environment. However, due to the fact that traditional bioaerosol collection instruments or device counterpart detection methods are mainly suitable for bacteria and fungal pathogens, the monitoring of virus aerosols has been limited, and the monitoring of viruses is inconvenient to operate and has poor sensitivity, which is seriously unsuitable for the current field of infectious diseases The current monitoring research status of the virus restricts the monitoring requirements of pathogens in the environment, and the new virus sampler has become one of the current research hotspots in this field.

Since viruses are non-cellular microorganisms that must rely on cells to replicate and multiply, the particles are small (nanoscale), and air dilution leads to low concentrations, it has always been a problem to collect respiratory viruses in ambient air to a detectable level. Obviously, viral air collectors should focus on the aerosol particles produced by breathing. Studies have shown that most of the aerosol particles produced by human respiratory activities (including coughing, sneezing, talking and breathing, etc.) are smaller than 1 μm (Edwards et al, 2004 ), while influenza viruses associated with respiratory activity are mainly distributed in 0.3-1μm aerosol particles (Fabian et al, 2008). Therefore, the existing air samplers have high collection efficiency for particles larger than 1 μm, but low efficiency for collecting virus aerosol particles, and lack of pertinence is one of the main reasons. The vast majority of existing bioaerosol samplers are not specifically designed for viral aerosols, and the collection spectrum is wide, which is not suitable for collecting virus samples. The typical reasons can be summarized as follows:

l) The flow rate of the sampler is small or the working time is short, it is difficult to collect a sufficient amount of air samples to ensure the minimum limit of sufficient virus detection;

2) The collection efficiency of aerosol particles is low, especially the collection efficiency of aerosol particles at the submicron level;

3) Insufficient particle removal and concentration techniques in collection media lead to inefficiencies;

4) The technical problem of insufficient applicability of the collection medium for subsequent detection methods;

5) Poor operability for virus monitoring, or lack of on-site portability settings.

The latest foreign comparative analysis of virus collection on important commercial air samplers shows (Fabian et al, Indoor Air 2009, 19: 433-441), including SKC biosamper biological sampler, Teflon air filter, gelatin filter and CCI impact collection A total of four key commercial biological samplers were compared and analyzed, and the conclusions are: new aerosol virus samplers need to be studied; existing commercial biological samplers lack efficiency in virus collection and infectivity maintenance; Focus on aspects of collection efficiency and collection liquid media optimization at the particle size stage of viral aerosols. Except that some foreign military units have carried out monitoring and detection of viral aerosols based on the needs of anti-bioterrorism, there are no air virus monitoring and enrichment instruments specially used in sensitive places such as large gatherings, confined spaces of vehicles or medical institutions, especially For the development of nanoscale virus sampling and monitoring devices and supporting virus concentration to form highly sensitive and specific detection devices, there are still many requirements for improvement in terms of sampling efficiency, concentration efficiency and simplicity, and there is still a long way to go before the actual application value.

At present, the air microbial samplers used in environmental monitoring in the domestic public health field can be roughly divided into three categories: air microorganisms captured on solid medium or other adhesive surfaces, collected in nutrient solution, collected on filter membranes or other filter materials In terms of other aspects, this type of commercial air microbial sampler is mainly aimed at collecting microbial aerosols such as micron-sized bacteria, and the collection efficiency of sub-micron level particles is low. Due to the characteristics of nano-scale particles of viruses and the high variability of pathogens such as influenza viruses, the concentration and discharge of viruses in the local respiratory tract of different viruses are different, and the detection methods of different viruses are different. There is a lack of systematic and targeted research, and the corresponding sample collection devices do not have the integration of virus enrichment technology. The sensitivity of virus detection needs to be improved, and the connection with subsequent detection technologies also needs to be improved. No air sampling device that integrates virus collection and enrichment has been seen. . At the same time, based on the environmental requirements of air virus monitoring, the existing biological samplers lack portability, simplicity and specificity in terms of the characteristics of relatively closed and narrow spaces such as vehicles, biosafety laboratories or animal farms, making sampling The device does not meet the actual requirements.

Contents of the invention

In order to solve the problems existing in the prior art, the present invention provides a device for collecting and enriching viral aerosols, especially an integrated device for effectively collecting and integrating virus enrichment of bioaerosol virus samples in ambient air .

To achieve the above object, the present invention provides a viral aerosol collection and enrichment instrument comprising a frame, a gas supply device, a gas outlet unit nozzle and a collection unit tube; the gas supply device is used to collect gas samples to be detected, and through The gas delivery pipe inputs the gas sample into the nozzle of the gas outlet unit; the nozzle of the gas outlet unit and the pipe of the collection unit are both arranged on the frame, and the pipe of the collection unit is detachably arranged under the nozzle of the gas outlet unit, and the nozzle of the gas outlet unit is set in it. There is a test liquid; the gas sample is rotated and sprayed into the tube of the collection unit through the nozzle of the gas outlet unit, and the aerosol contained in the gas to be tested is absorbed by the test liquid therein.

Further, the spray head of the air outlet unit is a cylindrical chamber, and several air outlets are arranged on its bottom surface. Spatially spaced and obliquely downward at a set angle with respect to said axis.

Further, the number of the jet outlets is greater than or equal to 3.

Further, the bottom surface is a downwardly protruding surface.

Further, the several air jet outlets are located at the same height in the vertical direction.

Further, the collection unit tube is a container composed of a cylinder and a conical cylinder arranged below the cylinder, and the volume of the conical cylinder part is greater than or equal to 15ml.

Further, the collection and enrichment device also includes an enrichment unit frame, the enrichment unit frame includes a tube frame capable of free rotation in the horizontal and vertical directions and an elastic spring for fixing the collection unit tube arranged on the tube frame. Fix the circlip; after the collection unit tube support device is tested, the collection unit tube is set on the enrichment unit frame.

Further, the enrichment unit frame further includes a magnetic device arranged at the bottom of the tube frame.

Further, the outer wall of the collection unit tube is provided with a bayonet matching with the elastic fixing ring.

Further, the test solution is a test solution with virus immune magnetic beads; and the virus carried by the gas to be tested is captured by the virus immune magnetic beads.

The principle of the viral aerosol collection and enrichment instrument defined in the present invention is to collect aerosol by using gas-liquid mixing, and put immunomagnetic beads into the test solution to capture the virus carried by the gas to be tested. The device consists of two important parts, the nozzle of the air outlet unit and the tube of the collection unit, which are combined to form a complete air sampling and virus enrichment instrument. The structure is simple, the operation links are coherent, and the implementation is simple. , especially when docked with commonly used virus molecular biology detection methods, it can directly enter the nucleic acid extraction and detection steps of pathogens without additional processing, saving time and effort. In addition, the three main components of the air outlet unit nozzle, the collection unit tube and the enrichment unit frame in the present invention can also be used independently to complete the independent functions of each part.

Description of drawings

Fig. 1 is the schematic diagram of the structure of the collection and enrichment instrument of the present invention;

Fig. 2 is a structural schematic diagram of a nozzle of an air outlet unit;

Fig. 3 is the structural schematic diagram of the nozzle of the second air outlet unit;

Fig. 4 is a structural schematic diagram of another nozzle of an air outlet unit;

Fig. 5 is a schematic diagram of the structure of the collection unit tube;

Fig. 6 is a sectional view of the connection between the collection unit pipe and the nozzle of the air outlet unit;

Fig. 7 is a schematic diagram of the framework structure of the enrichment unit.

Detailed ways

As shown in FIG. 1 , the viral aerosol collection and enrichment instrument includes a frame 1 , an air supply device, an air outlet unit nozzle 3 , a collection unit tube 4 and an enrichment unit rack 5 . And the up and down directions are set according to the use state of the collection and enrichment device, that is, the collection unit tube 4 is located below the nozzle 3 of the air outlet unit. The gas supply device includes an air pump and a gas delivery pipe 2, and the gas delivery pipe 2 transports the gas to be measured drawn in by the air suction pump to the nozzle 3 of the gas outlet unit, and then sprays out from the gas injection outlet 7. The nozzle 3 of the air outlet unit is a cylindrical chamber, and its bottom surface 6 is preferably an outwardly protruding surface, which can be a part of a spherical surface or a regular or irregular curved surface or plane such as a conical surface or a pyramidal surface; as shown in Figure 2-4 The bottom surface 6 of the air outlet unit nozzle 3 is also provided with some air outlets 7, which are evenly distributed with respect to the axis of the air outlet unit nozzle 3, and are preferably positioned at the same height in the vertical direction, and the axes of some air outlets 7 are uniform. It is spaced apart from the axis of the spray head 3 of the air outlet unit, and is set at an obliquely downward angle relative to the axis. The number of air jet outlets 7 is not less than 3, preferably 3-6. The advantages of this arrangement of the gas jet outlet 7 are as follows: the gas jet outlet 7 is directly processed on the bottom surface, which is convenient for processing, and can ensure that the jetting direction of the gas remains unchanged during the jetting process. The number of jet outlets is large, the jet outlet pipe is short, not easy to be blocked, and easy to clean; the jet airflow evenly forms a tangential airflow in the downward direction of the same surface, forming a fully rotating downward driving force, driving the liquid level to rotate and rise, and around the jet outlet A semi-circular wrapping liquid surface is formed to realize complete gas-liquid mixing and comprehensive collection of aerosols.

As shown in Figures 5 and 6, the collection unit tube 4 is a container composed of a cylinder 8 and a conical cylinder 9 arranged below the cylinder 8. The upper end of the cylinder 8 is open, and the lower end is connected to the large end of the conical cylinder 9. The top of the inner wall of the cylinder 8 is provided with a screw port so as to be connected with the spray head 3 of the air outlet unit, or to be detachably connected with the spray head 3 of the air outlet unit by other means. The outer wall of the collection unit tube 4 is also provided with an inward shallow concave bayonet 10, which is used to fix the collection unit tube on the enrichment unit frame 5. The bayonet is preferably arranged on the cylinder 8 and the conical cylinder 9. Junction. The conical cylinder part of the collection unit tube is mainly used to store the collection solution, and the liquid capacity of the conical cylinder part is not less than 15ml. The structure of the collection unit tube 4 can ensure enough test liquid to fully mix with a large amount of air samples, otherwise effective gas-liquid mixing will not be achieved or aerosol loss will occur due to the rapid evaporation of the test liquid. The test solution can be a test solution with virus immune magnetic beads; and the virus carried by the gas to be tested is captured by the virus immune magnetic beads.

As shown in FIG. 7 , the enrichment unit frame 5 is a tube frame that can rotate freely in the horizontal and vertical directions, and an elastic fixing spring 11 is provided on the top of the tube frame for docking with the bayonet 10 of the collection unit tube. The elastic fixing circlip 11 can realize the fixing of the collection unit tube, and can also manually pick and place the collection unit tube 4 at any time. A magnetic device 12 is provided at the bottom of the tube rack, which can be a fixed permanent magnet, etc., and the size of the tube rack needs to match the collection unit tube 4 . As shown in FIG. 1 , the enrichment unit frame 5 is arranged at the lower end of the frame 1 , and can be received into the frame 1 by rotating the tube frame horizontally, or rotated out during use, so as to fix the collection unit tube 4 . The liquid in the collection unit tube 4 can be poured out by rotating the tube holder in the vertical direction. In addition, the enrichment unit frame 5 can also be set independently from the frame 1, but its structure and function remain unchanged.

During the test, the viral aerosol collection and enrichment instrument of the present invention is set in the region to be tested, the air pump is started, the gas to be tested is sucked in, and is input into the gas outlet unit nozzle 3 through the gas delivery pipe 2, and is set Some jet outlets 7 on its bottom surface 6 are ejected into the test liquid in the collection unit pipe 4, because some jet outlets 7 are positioned at the same height of the bottom surface 6, and their outlets are all facing obliquely downwards, so the jets are ejected by it. The gas to be tested is sprayed into the collection unit tube 4 at the same time to form a rotating air flow, which drives the test liquid in the collection unit tube 4 to rotate, so that the liquid level rises, and realizes wrapping the curved surface of the liquid injected into the air flow, so that all collected The gas to be tested can be completely sprayed onto the surface of the test liquid and continuously mixed in the rotating test liquid, thereby achieving efficient mixing of air and test liquid and effective capture and collection of aerosols in the air. After the gas collection is completed, the collection unit tube 4 is removed from the nozzle 3 of the gas outlet unit, and the collection unit tube 4 is fixedly placed on the enrichment unit frame 5 to stand still, and the magnetic beads in the liquid to be tested are all adhered by magnetic force After collecting the bottom of the unit tube 4, rotate the tube rack vertically to pour out the test solution, and keep the virus-enriched magnetic beads at the bottom of the tube for further virus detection.

The advantages of the viral aerosol collection and enrichment instrument of the present invention are as follows: 1. The design of the nozzle of the air outlet unit conforms to the principle of the physical cyclone, and the collected air flow and the subsequent arc-shaped wrapping liquid surface can realize full contact between gas and liquid, ensuring that the collected The aerosol particles of different particle sizes contained in the gas to be tested are fully collected by the test liquid; 2. The design and layout of several gas outlet unit nozzles in the same cavity and short pipelines make the gas outlet unit nozzles not easy to block and easy to clean; 3. The collection unit tube It is a combined shape of cylinder and cone. On the one hand, it can ensure that after the airflow drives the liquid surface to rotate, the upper space of the liquid forms a large arc-shaped concave surface, so as to fully wrap the gas injection port and ensure effective gas collection and mixing. On the other hand, The pointed design at the lower end of the cone is beneficial to accommodate the small volume of the subsequent concentrated magnetic beads, which is convenient for recovery; 4. The collection unit tube has a large liquid sample capacity, which can ensure the collection of large air flow after the airflow is driven and the effective mixing with the test solution , Virus capture immune magnetic beads are directly added to the test solution to capture the virus in the aerosol while the gas and liquid are mixed; 5. The enrichment unit frame can be folded horizontally and vertically, which is convenient for integration into the instrument device. The bottom end of the frame is a fixed permanent magnet, which is convenient to contact with the conical bottom end of the conical part of the collection unit tube to enrich the magnetic beads in the tube.

Claims (9)

1. a viral aerosol gathers enriching instrument, it is characterized in that, described collection enriching apparatus comprises framework, air feeder, the unit spray head of giving vent to anger and collecting unit pipe; Described air feeder is used for collecting gas sample to be detected, and by pneumatic tube, described gas sample is input in the unit spray head of giving vent to anger; Described unit spray head and the collecting unit Guan Jun of giving vent to anger is arranged on framework, and described collecting unit pipe dismountable below that is arranged on the described unit spray head of giving vent to anger is provided with test fluid in it; Described gas sample is injected in described collecting unit pipe by the unit spray head rotation of giving vent to anger, and absorbs contained aerosol in gas to be measured by described test fluid wherein; Described collection enriching apparatus also comprises enrichment unit frame, and described enrichment unit frame comprises can be realized the pipe support that the horizontal and vertical direction rotates freely and be arranged on the flexible fastening jump ring that is used for fixing described collecting unit pipe on pipe support.

2. gather enriching instrument according to viral aerosol described in right 1, it is characterized in that, the described unit spray head of giving vent to anger is cylindrical chamber, be provided with some jet outlets on its bottom surface, some described jet outlets are uniform with respect to the axis of the unit spray head of giving vent to anger, the axis of some jet outlets all spatially is separated by with the axis of the unit spray head of giving vent to anger, and with respect to described axis to oblique below inclination set angle.

3. viral aerosol according to right 2 gathers enriching instrument, it is characterized in that, the quantity of described air outlet is more than or equal to 3.

4. gather enriching instrument according to viral aerosol described in right 2, it is characterized in that, described bottom surface is the face that protrudes downwards.

5. gather enriching instrument according to viral aerosol described in right 2, it is characterized in that, described some jet outlets are positioned at sustained height in vertical direction.

6. gather enriching instrument according to viral aerosol described in right 1, it is characterized in that, described collecting unit pipe is for by cylinder be arranged on the container that the taper cone barrel of described cylinder below consists of, and the capacity of described taper cone barrel part is more than or equal to 15ml.

7. gather enriching instrument according to viral aerosol described in right 1, it is characterized in that, described enrichment unit frame also comprises the magnetic devices that is arranged on described pipe support bottom.

8. gather enriching instrument according to viral aerosol described in right 1, it is characterized in that, be provided with the bayonet socket that matches with described flexible fastening jump ring on the outer wall of described collecting unit pipe.

9. gather enriching instrument according to viral aerosol described in right 1, it is characterized in that, described test fluid is the test fluid with the virus immunity magnetic bead; And catch by described virus immunity magnetic bead the virus that gas to be measured carries.

Images