CN102175493B - Expiratory air sampling box and method - Google Patents
Expiratory air sampling box and method Download PDFInfo
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- CN102175493B CN102175493B CN201110020115A CN201110020115A CN102175493B CN 102175493 B CN102175493 B CN 102175493B CN 201110020115 A CN201110020115 A CN 201110020115A CN 201110020115 A CN201110020115 A CN 201110020115A CN 102175493 B CN102175493 B CN 102175493B
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Abstract
The invention discloses a simple and efficient expiratory air sampling box and a method for sampling expiratory air by using the sampling box. The sampling box comprises a sampling box base (1) and a sampling box cover (4), wherein the sampling box cover is provided with an expiratory air inlet (5); an ice layer (2) is arranged on the sampling box base; and a super-hydrophobic film (3) is arranged on the ice layer. The sampling method comprises the following steps of: conveying expiratory air of a sampler to the surface of the super-hydrophobic film treated at low temperature; condensing the expiratory air on the super-hydrophobic film to form small liquid drops; dripping a certain amount of water which does not contain nucleic acid onto the surface of the super-hydrophobic film by using a liquid-transferring gun; stirring the liquid drops by using a gun head of the liquid-transferring gun to make sure the contact and rolling of the liquid drops on the surface of the super-hydrophobic film; and transferring the liquid drops into a sterile centrifugal tube to finish sampling. By adopting the sampling box and the method, the defects such as heavy weight, large size, low repeated utilization ratio and the like of the conventional expiratory air sampler are overcome, and portability and reutilization are realized. The sampling box and the method can be applied to the aspects of detection of clinical diseases, prevention and control of flu epidemic situation and the like.
Description
Technical field
The present invention relates to a kind of simple and easy, the expiratory air sampling box and the method for sampling efficiently, be mainly used in clinical disease and detect, take precautions against control influenza epidemic situation etc.
Background technology
The outburst of the large-scale influenza of SARS in 2003 and H1N1 in 2009 has caused huge social fear and economic loss.When flu outbreak, usually adopt the body temperature primary dcreening operation, isolate and means such as make a definite diagnosis and take precautions against the further diffusion of influenza then., by the patient of influenza infection being arranged certain latent period, though there is not symptom clinically, also is the carrier of virus, has infectivity.Exhaled breath condensate (Exhaled breath condensate is called for short EBC) is used to diagnose the illness more and more, and research object comprises cell factor, hydrogen peroxide (H
2O
2), nitrite (nitrite) etc.A nearest research shows the patient for primary breast cancer, and the hydrogen peroxide in the expiratory air is obviously higher.In addition, research finds that also common influenza A virus is arranged in patient's expiratory air, like this for coming the diagnose infections crowd that theoretical foundation is provided through the biotic component that detects in the expiratory air.
Yet expiratory air condenser commonly used is like RTube collection system (Respiratory Research; Inc; Charlottesville is VA) with EcoScreen condenser (Erich Jaeger GmbH, W ü rzburg; Germany) exist complex structure, heaviness, efficient low, carry defectives such as inconvenience, these defectives have greatly limited exhaled breath condensate in medically application.
Summary of the invention
The object of the invention:, the invention provides a kind of portable simple and easy expiratory air sampling box and the method for sampling for overcoming shortcomings such as heavy, the huge and recycling rate of waterused of at present traditional expiratory air sampling thief is low.
Technical scheme of the present invention is following:
Scheme 1: a kind of expiratory air sampling box comprises: the sampling box base, sampling lid, said sampling box have covered expiratory air inlet, it is characterized in that, place ice sheet on the sampling box base, place the super-hydrophobic film that a ultralow temperature is handled on the ice sheet.Said ultralow temperature is-70 ℃.
Scheme 2: a kind of preferred realization as scheme 1 is characterized in that said sampling lid and sampling box base seamless combination.For example, can use modes such as spiral shell button or buckle to carry out snapping.Can guarantee that like this anhydrous steam pollutes in sampling process.
Scheme 3: a kind of preferred realization as scheme 1 is characterized in that the material of said sampling box is a teflon.Can reduce expiratory air in the inner absorption of device.
Scheme 4: as a kind of preferred realization of scheme 1; It is characterized in that; It is rectangular parallelepiped that said sampling box is shaped as the bottom; Top is truncated rectangular pyramids, and the length and width height of said sampling box base is of a size of 100mm * 50mm * 30mm, sampling box covers, the bottom is of a size of 50mm * 50mm * and 25mm and 100mm * 50mm * 25mm.
Scheme 5: a kind of preferred realization as scheme 1 or 3 or 4 is characterized in that the thickness of said sampling box box wall is 3mm.Whole like this sampling box small-sized, weight can be accomplished less than 50g, and is very portable.
The present invention provides the method for utilizing scheme 1 described sampling thief to carry out the expiratory air sampling simultaneously, as follows:
Scheme 6: a kind of method of utilizing scheme 1 described sampling box to carry out the expiratory air sampling, it is characterized in that, comprise the steps:
A) one deck sterilized water is joined on the sampling box base;
B) the sampling box base is carried out ultralow temperature and handle, be condensed into solid-state until said sterilized water;
C) super-hydrophobic film after the aseptic ultralow temperature (70 ℃) of suitable size is handled is positioned in the sampling box base on ice;
D) an aseptic wireway is inserted in the expiratory air inlet, makes wireway lower end and super-hydrophobic film keep a determining deviation;
E) the quilt person of adopting of expiratory air is put into mouth on the wireway, and is air-breathing with nose, with mouth expiratory air blown in the wireway again;
F) carried out repeatedly a last step, reaching predetermined number of times or after the fixed time, stopping sampling;
G) under gnotobasis, super-hydrophobic film is taken out;
H) use liquid-transfering gun to drip a certain amount of water that does not contain nucleic acid, stir the liquid pearl, make it on the super-hydrophobic film surface, can both have contact also to roll back and forth with the rifle head of liquid-transfering gun on the super-hydrophobic film surface;
I) the liquid pearl is drawn and transfers in the aseptic centrifuge tube with liquid-transfering gun, the liquid of collecting promptly contains the composition of expiratory air, and sampling finishes.
Scheme 7: a kind of preferred realization as scheme 6 is characterized in that in the said step b), said ultralow temperature is treated to a sampling box base and places ultra low temperature freezer to preserve.
Scheme 8: a kind of preferred realization as scheme 6 is characterized in that in the said step b), the spacing of wireway lower end and super-hydrophobic film is 2mm.Notice that the wireway lower end necessarily can not contact with super-hydrophobic film.
Scheme 9: a kind of preferred realization as scheme 6 is characterized in that in the said step f), the fixed time is 5-10 minute.
Scheme 10: a kind of preferred realization as scheme 6 is characterized in that said step h) in, the said amount that does not contain the water of nucleic acid is 5-10 μ L.
Beneficial effect of the present invention: device provided by the invention adopts simple super-hydrophobic film and special miniature sampling box to realize quick, the efficient sampling to patient expired gas, and gross weight is less than 50 grams.Principal feature is:
(1) utilizes the super-hydrophobic film after ultralow temperature is handled not only can realize quick, high-efficiency condensation, and realized convenient sampling, shortened the breathing sampling time greatly condensed fluid to expiratory air;
(2) utilize special expiratory air sampling box, can prevent loss, the cross pollution of expiratory air;
(3) this expiratory air device is simple and easy, cheap, reusable.
Description of drawings
Fig. 1 expiratory air easy device structural representation.
The PRELIMINARY RESULTS of the gene magnification (PCR) of total bacterium in Fig. 2 expiratory air sample.
Embodiment
Design of device: as shown in Figure 1, the expiratory air harvester mainly comprises sampling box base 1, sampling lid 4, expiratory air inlet 5 and ice sheet 2, super-hydrophobic film 3 compositions.The length and width height of sampling box base is of a size of 100mm * 50mm * 30mm.Sampling box covers, the following 50mm of being of a size of * 50mm * 25mm and 100mm * 50mm * 25mm, on the end face of lid, is drilled with the through hole that diameter is about 4mm, is the expiratory air inlet.The thickness of total contacts with lid closely in order to ensure the sampling box base about 3mm, has made the groove of 2mm on the border that sampling box base 1 contacts with sampling lid 4; Correspondingly, made the prominent groove of 2mm on the border of sampling lid 4.What the material of whole device used is teflon, can reduce expiratory air in the inner absorption of device.As long as do not hinder portable sampling, the size and dimension of harvester can according to circumstances change.
Gordian technique: because used super-hydrophobic film has very strong hydrophobic effect; Expiratory air becomes globule shape very soon on the super-hydrophobic film of handling through ultralow temperature; And there is not loss; Use liquid-transfering gun head (water that contains micro-free nucleic acid) on super-hydrophobic film, to drag, the automatic like this expiratory air that will be condensate on the super-hydrophobic film forms a big globule, thereby obtains the high-level efficiency sampling of expiratory air; Adopt the seal box technology to prevent the pollution of indoor water vapor simultaneously.
Operation instruction: at first sterilized water is joined on the sampling box base, make water depth reach 5mm and get final product, put into-70 ℃ of ultra low temperature freezers (Thermo Fisher Scientific Co.Marietta, OH) the middle preservation.Treat that sterilized water in the base has been condensed into when solid-state, (Parafilm Co.Menasha WI) is made into length and width and is of a size of 80mm * 40mm, is positioned in the sampling box base on ice with aseptic super-hydrophobic film.The lid of will sampling covers on the sampling box base, and is promptly as shown in Figure 1.An aseptic wireway (diameter is 4mm) is inserted in the expiratory air inlet, and wireway lower end and super-hydrophobic film keep the spacing about 2mm, and the lower end does not contact the upper surface of film.The quilt person of adopting of expiratory air is put into mouth on the wireway, and is air-breathing with nose, with mouth expiratory air blown to sampling box again; Be transported to the surface of super-hydrophobic film through wireway, because the interior environment temperature of film surface and box body is in subzero, expiratory air condenses on the super-hydrophobic film very soon; Form small liquid pearl (naked eyes are invisible), this expiratory air sampling operation continued after 5-10 minute, under gnotobasis; Super-hydrophobic film is taken out, use liquid-transfering gun (Eppendorf, Hauppauge; NY) Dropwise 5-10 μ L does not contain the water of nucleic acid on film surface, stirs the liquid pearl with the rifle head of liquid-transfering gun, makes it that contact and rolling back and forth can both be arranged on the super-hydrophobic film surface; Behind this end of operation, the liquid pearl is drawn and transfers in the aseptic centrifuge tube with liquid-transfering gun, the liquid of collecting promptly contains the composition of expiratory air.
Fig. 2 shows is the gene magnification figure of total bacterium in the expiratory air that 4 laboratory volunteers are sampled; Sample representes to utilize the laboratory participant's that this patent samples exhaled breath condensate among the figure; Wherein sampling film is blank, is under kindred circumstances but the blank sample that obtains when not sampling expiratory air.As can be seen from the figure, to four bit test persons, have in three the expiratory air to detect a certain amount of DNA of bacteria, and blank sample is negative.
The sampling thief provided by the invention and the method for sampling have overcome shortcomings such as heavy, the huge and recycling rate of waterused of present traditional expiratory air sampling thief is low, have realized portability.The expiratory air sample that these apparatus and method collect can supply each item biochemical analysis, is a kind of noninvasive sample method of sampling and device, and this is easy and simple to handle, with low cost.The present invention can be used for medical experiment, and the sample of sampling can supply the early detection and the diagnosis of follow-up disease.
Claims (5)
1. a method of utilizing the expiratory air sampling box to carry out the expiratory air sampling is characterized in that, comprises the steps:
A) one deck sterilized water is joined on the sampling box base (1);
B) sampling box base (1) is carried out ultralow temperature and handle, be condensed into solid-state until said sterilized water;
C) it is interior on ice that the super-hydrophobic film of the aseptic ultralow temperature of suitable size being handled (3) is positioned over sampling box base (1);
D) an aseptic wireway is inserted in the expiratory air inlet, makes wireway lower end and super-hydrophobic film keep a determining deviation;
E) the quilt person of adopting of expiratory air is put into mouth on the wireway, and is air-breathing with nose, with mouth expiratory air blown in the wireway again;
F) carried out repeatedly a last step, reaching predetermined number of times or after the fixed time, stopping sampling;
G) under gnotobasis, super-hydrophobic film is taken out;
H) use liquid-transfering gun to drip a certain amount of water that does not contain nucleic acid, stir the liquid pearl, make it on the super-hydrophobic film surface, can both have contact also to roll back and forth with the rifle head of liquid-transfering gun on the super-hydrophobic film surface;
I) the liquid pearl is drawn and transfers in the aseptic centrifuge tube with liquid-transfering gun, the liquid of collecting promptly contains the composition of expiratory air, and sampling finishes;
Described expiratory air sampling box; Comprise: sampling box base (1), sampling lid (4) has expiratory air inlet (5) on the said sampling lid (4); Sampling box base (1) is gone up and is placed ice sheet (2), places the super-hydrophobic film (3) that a ultralow temperature is handled on the ice sheet (2).
2. the method for claim 1 is characterized in that, in the said step b), said ultralow temperature is treated to a sampling box base (1) and places ultra low temperature freezer to preserve.
3. the method for claim 1 is characterized in that, in the said step b), the spacing of wireway lower end and super-hydrophobic film is 2mm.
4. the method for claim 1 is characterized in that, in the said step f), the fixed time is 5-10 minute.
5. the method for claim 1 is characterized in that, said step h) in, the said amount that does not contain the water of nucleic acid is 5-10 μ L.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201110020115A CN102175493B (en) | 2011-01-18 | 2011-01-18 | Expiratory air sampling box and method |
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|---|---|---|---|
| CN201110020115A CN102175493B (en) | 2011-01-18 | 2011-01-18 | Expiratory air sampling box and method |
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| CN102175493A CN102175493A (en) | 2011-09-07 |
| CN102175493B true CN102175493B (en) | 2012-08-29 |
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| DE10232849A1 (en) * | 2002-07-19 | 2004-02-12 | Abb Patent Gmbh | Gas analyzer, e.g. for field detection of odorless, invisible toxic agents and pollutants, has self-contained sampling and detector, and sample is transferred to detector under action of force field |
| US7914460B2 (en) * | 2006-08-15 | 2011-03-29 | University Of Florida Research Foundation, Inc. | Condensate glucose analyzer |
| US8394030B2 (en) * | 2009-03-18 | 2013-03-12 | Carefusion 207, Inc. | Exhaled breath condensate biometric marker measurement apparatus and method |
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Non-Patent Citations (2)
| Title |
|---|
| Cam.T.V.Anh et al..《A hydrogen peroxide sensor for exhaled breath measurement》.《Sensors and Actuators B:Chemical》.2005,第111-112卷第494-499页. * |
| Irfan Rahman, Saibal K. Biswas.《Non-invasive biomarkers of oxidative stress: reproducibility and methodological issues》.《Redox Report》.2004,第9卷(第3期),第127-128页. * |
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Effective date of registration: 20181105 Address after: 100085 F block 606, 9 on the 3rd Street, Haidian District, Beijing. Patentee after: Beijing Zhong LAN Technology Co.,Ltd. Address before: 100871 No. 5, the Summer Palace Road, Beijing, Haidian District Patentee before: Peking University Effective date of registration: 20181105 Address after: 100085 D405, block D, 9 Sandi street, Haidian District, Beijing. Patentee after: Beijing Ding LAN Technology Co.,Ltd. Address before: 100085 F block 606, 9 on the 3rd Street, Haidian District, Beijing. Patentee before: Beijing Zhong LAN Technology Co.,Ltd. |
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