CN111024661B - Fluorescence detection card and detection method for environmental pollutants - Google Patents
Fluorescence detection card and detection method for environmental pollutants Download PDFInfo
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- CN111024661B CN111024661B CN201911277096.5A CN201911277096A CN111024661B CN 111024661 B CN111024661 B CN 111024661B CN 201911277096 A CN201911277096 A CN 201911277096A CN 111024661 B CN111024661 B CN 111024661B
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Abstract
The invention provides a fluorescent detection card for environmental pollutants, and relates to the technical field of pollutant detection. The utility model provides an environmental contaminant's fluorescence detection card, includes first fluorescence pick-up plate, first detection window, and first fluorescence pick-up plate articulates there is second fluorescence pick-up plate, and second fluorescence pick-up plate is equipped with the second and detects the groove, has solved current detection card structure simple relatively, leads to fluorescence detection card to use the problem that the limitation is big. A method for detecting environmental pollutants by using a fluorescence detection card comprises the steps of adjusting the overlapping of a first fluorescence detection plate and a second fluorescence detection plate, adding a reagent, adding a detection sample and carrying out photoelectric treatment, and can realize the fluorescence detection of a small number of groups of information of the environmental pollutants. A method for detecting environmental pollutants by using a fluorescence detection card comprises the steps of adjusting a first fluorescence detection plate and a second fluorescence detection plate to be parallel, adding a reagent, adding a detection sample and carrying out photoelectric treatment, and can realize the fluorescence detection of information of a large number of groups of environmental pollutants.
Description
Technical Field
The invention relates to the technical field of pollutant detection, in particular to a fluorescent detection card and a fluorescent detection method for environmental pollutants.
Background
The environmental pollutants refer to substances which change the normal composition and properties of the environment after entering the environment, are directly or indirectly harmful to human survival or cause the decline of the natural ecological environment, and are the objects of environmental monitoring and research. Most environmental pollutants are produced by human production and living activities. Some substances are originally useful substances in production, even nutrient elements necessary for people and organisms, and are discharged in large quantities due to underutilization, so that not only is the resource waste caused, but also the substances can become environmental pollutants.
The environmental pollutants are mainly organic pollutants, and the current industrial detection methods for the organic pollutants (organic matters such as polycyclic and squaric hydrocarbons) mainly comprise two main types of chemistry and biology: the former is mostly chromatography and mass spectrometry, the instrument and equipment are large and expensive, the detection item is single, the flux is small, the sample pretreatment is complex, and new unknown substances in the sample cannot be found out; for some aldehyde compounds, the aldehyde compounds and dyes form colored compounds to be detected by a visible light spectrophotometry, the sensitivity is low, and the detection items are very limited. Biological detection methods, such as uterus growth experiments, hepatocyte vitellogenin production experiments, human breast cancer cell (MCF-7) proliferation experiments, recombinant gene yeast experiments, yeast two-hybrid experiments and other five main environmental estrogen biological detection methods, the instruments need an enzyme labeling instrument, an animal cell incubator, an ultra-clean bench and a related ultra-clean working environment, the method is complicated, the time is long, the cost is high, and the detection sample is single. These methods are not suitable for rapid detection at the basement level and in the field.
In order to increase the detection quality of industrial environmental organic pollutants, a fluorescence detection mode appears, wherein the fluorescence detection is a natural luminescence reaction, and human cells, bacteria, mould and food residues can be detected by reacting luciferase with ATP, and a reaction result can be obtained within 15 seconds. The illuminance is measured by a dedicated device and is represented in digital form. In fluorescence detection, a fluorescence detection card is filled with a non-cell expression system reagent and a detection sample, and the fluorescence detection card is combined with a photoelectric technology detection means (the fluorescence detection card and the photoelectric technology detection means form the conventional organic pollutant detector) to detect the fluorescence intensity of expressed GFP (green fluorescent protein) so as to judge the concentration of organic matters in the organic matter detection sample and evaluate the degree of environmental pollutants. The existing fluorescent detection card has a relatively simple structure, so that the problem of large use limitation of the fluorescent detection card is caused.
Disclosure of Invention
The invention aims to provide a fluorescent detection card for environmental pollutants, which solves the problems that the existing detection card has a relatively simple structure and causes large use limitation of the fluorescent detection card.
The invention aims to provide a detection method for detecting environmental pollutants by adopting a fluorescence detection card, which can realize the fluorescence detection of a few groups (within three groups) of information of the environmental pollutants.
The invention aims to provide a detection method for detecting environmental pollutants by adopting a fluorescence detection card, which can realize the fluorescence detection of information of more groups (within six groups) of environmental pollutants.
The embodiment of the invention is realized by the following steps:
the utility model provides an environmental contaminant's fluorescence detection card, including first fluorescence pick-up plate, be equipped with three first detection groove on the first fluorescence pick-up plate, the bottom in first detection groove is equipped with the first detection window of detachable, one side of first fluorescence pick-up plate articulates there is the second fluorescence pick-up plate rather than overlapping, and first fluorescence pick-up plate is the same with second fluorescence pick-up plate shape, one side of second fluorescence pick-up plate is equipped with the second that can overlap with first detection groove and detects the groove, the bottom in second detection groove is equipped with detachable second and detects the window, first detection window, the second detects the window and makes by transparent material.
In some embodiments of the present invention, the first fluorescence detection plate and the second fluorescence detection plate are made of plastic, the three first detection grooves are coaxially arranged on the first fluorescence detection plate, the three second detection grooves are coaxially arranged on the second fluorescence detection plate, and the inner sides of the first detection groove and the second detection groove are made of transparent materials.
In some embodiments of the invention, the first fluorescence detection plate and the second fluorescence detection plate are strip-shaped, opposite sides of the first fluorescence detection plate and the second fluorescence detection plate are hinged through hinges, and the first fluorescence detection plate and the second fluorescence detection plate are connected through a lock catch at the side far away from the hinged part.
In some embodiments of the invention, the first fluorescence detection plate and the second fluorescence detection plate are detachably connected by a lock catch at a side far away from the hinge joint, an annular groove is arranged in the upper side of the second detection groove, a projection butted with the annular groove is arranged on the upper side of the first detection groove, and the projection is made of transparent material.
In some embodiments of the invention, one side of the first detection window is hingedly connected to the first fluorescence detection plate, the other side of the first detection window is detachably connected to the first fluorescence detection plate, one side of the second detection window is hingedly connected to the second fluorescence detection plate, and the other side of the second detection window is detachably connected to the second fluorescence detection plate.
In some embodiments of the invention, a clamping groove is arranged on one side of the first fluorescence detection plate facing the first detection window, a spherical limiting groove capable of being in coaxial line butt joint with the clamping groove is arranged on one side of the first detection window away from the hinged part, a spring in a compressed state is arranged in the clamping groove, a spherical limiting body capable of being in butt joint with the spherical limiting groove is connected to one side of the spring, and the spherical limiting body cannot fall off from the clamping groove.
In some embodiments of the invention, a clamping groove is arranged on one side of the second fluorescence detection plate facing the second detection window, a spherical limiting groove capable of being in coaxial line butt joint with the clamping groove is arranged on one side of the second detection window away from the hinged part, a spring in a compressed state is arranged in the clamping groove, a spherical limiting body capable of being in butt joint with the spherical limiting groove is connected to one side of the spring, and the spherical limiting body cannot fall off from the clamping groove.
The first detection window and the second detection window are both in a threaded column shape, the screw rod parts of the first detection window and the second detection window which are in the threaded column shape are cylindrical, and the first detection window and the second detection window are respectively connected with the first detection groove and the second detection groove in a threaded mode.
A detection method for detecting environmental pollutants by adopting a fluorescence detection card comprises the following steps:
adjusting: adjusting the first fluorescence detection plate and the second fluorescence detection plate to enable the first fluorescence detection plate and the second fluorescence detection plate to be overlapped, and opening three second detection windows on the second fluorescence detection plate;
adding a reagent: filling the three pairs of overlapped first detection grooves and second detection grooves with a testosterone comamonas non-cell system reagent for expressing 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase through three opened second detection windows;
adding a detection sample, namely adding environmental pollutants into the overlapped first detection groove and the second detection groove, incubating the environmental pollutants with a Comamonas testosteroni non-cell system reagent for expressing 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase at 37 ℃ for 10 minutes, and expressing the Comamonas testosteroni non-cell expression system reagent for expressing 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase to generate green fluorescent protein;
and (3) photoelectric treatment: by utilizing the principle of a photoelectric technology, the first fluorescence detection plate, the second fluorescence detection plate and a photoelectric technology detection means are organically combined to design an environmental dye detector, the fluorescence intensity of expressed green fluorescent protein is detected to judge the concentration of organic pollutants in the environmental pollutants, the degree of the environmental pollutants is evaluated, the Comamonas testosteroni capable of expressing 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase is replaced by a green fluorescent protein reporter gene through a transgenic technology to construct a Comamonas testosteroni fluorescent variant strain of the 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase, the variant strain can efficiently express the green fluorescent protein in the presence of the organic pollutants, and the strains are increased and crushed, obtaining various organelles in the thallus and green fluorescent protein reporter genes to prepare the transformed Comamonas testosteroni non-cell system reagent of the 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase.
A detection method for detecting environmental pollutants by adopting a fluorescence detection card comprises the following steps:
adjusting: adjusting the first fluorescence detection plate and the second fluorescence detection plate to enable the second fluorescence detection plate to be turned away from the first fluorescence detection plate and arranged side by side with the first fluorescence detection plate, and enabling the first detection window and the second detection window to be respectively in close fit with the bottoms of the first detection groove and the second detection groove;
adding a reagent: filling the first detection groove and the second detection groove with testosterone comamonas non-cell system reagents for expressing 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase through upper openings of the first detection groove and the second detection groove respectively;
adding a detection sample, namely respectively adding environmental pollutants into a first detection groove and a second detection groove, incubating the environmental pollutants with a Comamonas testosteroni non-cell system reagent for expressing 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase at 37 ℃ for 10 minutes, and expressing the Comamonas testosteroni non-cell expression system reagent for expressing 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase to generate green fluorescent protein;
photoelectric treatment: by utilizing the principle of the photoelectric technology, the first fluorescence detection plate, the second fluorescence detection plate and the photoelectric technology detection means are organically combined to design an environmental pollutant detector, the fluorescence intensity of expressed green fluorescent protein is detected to judge the concentration of organic pollutants in the environmental pollutants, the degree of the environmental pollutants is evaluated, the 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase gene in the Comamonas testosteroni can be replaced by a green fluorescent protein reporter gene through a transgenic technology, the 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase gene in the Comamonas testosteroni is constructed, the fluorescent variant strain of the Comamonas testosteroni of the 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase can be constructed, the variant strain can efficiently express the green fluorescent protein in the presence of the organic pollutants, and the strains are increased and crushed through a large amount of bacteria, obtaining various organelles in the thallus and green fluorescent protein reporter genes to prepare the transformed Comamonas testosteroni non-cell system reagent of the 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase.
The embodiment of the invention at least has the following advantages or beneficial effects:
the invention provides a fluorescent detection card for environmental pollutants, which comprises a first fluorescent detection plate, wherein three first detection grooves are arranged on the first fluorescent detection plate, the bottom of each first detection groove is a detachable first detection window, one side of the first fluorescent detection plate is hinged with a second fluorescent detection plate which can be overlapped with the first fluorescent detection plate, the first fluorescent detection plate and the second fluorescent detection plate are identical in shape, one side of the second fluorescent detection plate is provided with a second detection groove which can be butted with the first detection groove, the bottom of the second detection groove is a detachable second detection window, and the first detection window and the second detection window are both made of transparent materials. When environmental pollutants with less contrast data are detected and the second fluorescence detection plate is turned over to the upper side of the first fluorescence detection plate, the first detection groove is in butt joint with the second detection groove, the second detection window is detached, the first detection groove and the second detection groove are in an open state, when the first detection groove and the second detection groove are longitudinally overlapped, three groups of detection grooves (the first detection groove and the second detection groove are integrated) exist, the floor area of the fluorescence detection card is reduced, the fluorescence detection card is easy to store, meanwhile, the depth of the three groups of detection grooves (the first detection groove and the second detection groove are integrated) is large, and a large number of reagents and detection samples can be filled for detection. When detecting environmental pollutants with more contrast data, the second fluorescence detection plate is turned away from the upper side of the first fluorescence detection plate, the second fluorescence detection plate and the first fluorescence detection plate are arranged in parallel, the first detection window and the second detection window respectively seal one end port of the first detection groove and one end port of the second detection groove, detection samples and reagents with different quantities are respectively added into the six detection grooves (the three first detection grooves and the three second detection grooves), and detection of multiple data can be achieved. The switching of above-mentioned two kinds of detection methods (first detection groove overlaps and separates with the second detection groove), can realize the use of the different detection states of same fluorescence detection card, increased the application range of fluorescence detection card, reduced the use limitation of fluorescence detection card, simultaneously, first detection window, the connection dismantled of second detection window make the first both ends that detect the groove, the second detects the groove all be in and open the state, be convenient for wash first detection groove, the second detection groove.
After the first fluorescence detection plate and the second fluorescence detection plate are overlapped, three groups of detection grooves (the first detection groove and the second detection groove are integrated) can appear, detection samples and reagents with different quantities can be correspondingly added into the three groups of detection grooves, and then the detection of small-scale (within three groups) environmental pollutant samples can be realized through the photoelectric technology treatment.
After the first fluorescence detection plate is separated from the second fluorescence detection plate, three groups of first detection grooves and three groups of second detection grooves can appear, detection samples and reagents with different amounts can be correspondingly added into the three groups of first detection grooves and the three groups of second detection grooves, and then the detection of large-scale (within six groups) environmental pollutant samples can be realized through photoelectric technology processing.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
FIG. 1 is a schematic structural diagram of a fluorescence detection card for environmental pollutants according to an embodiment of the present invention;
FIG. 2 is a bottom view of FIG. 1;
FIG. 3 is a full sectional view of A in FIG. 2;
FIG. 4 is a side view of FIG. 1;
FIG. 5 is a partial schematic view of a first fluorescent detection panel overlapping a second fluorescent detection panel in accordance with an embodiment of the present invention;
FIG. 6 is a schematic structural diagram of a method for detecting environmental pollutants by using a fluorescence detection card according to an embodiment of the present invention;
FIG. 7 is a schematic view of the first and second fluorescence detection plates when they are not connected to the first and second detection windows according to the embodiment of the present invention.
Icon: 1-a first fluorescence detection plate, 2-a hinge, 3-a second fluorescence detection plate, 4-a second detection groove, 5-an annular groove, 6-a bulge, 7-a second detection window, 8-a first detection window, 9-a spherical limiting body, 10-a spring, 11-a clamping groove, 12-a spherical limiting groove, 13-a lock catch, 14-an environmental dye detector and 15-a first detection groove.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. The components of embodiments of the present invention generally described and illustrated in the figures herein may be arranged and designed in a wide variety of different configurations.
Thus, the following detailed description of the embodiments of the present invention, as presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. 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.
It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, it need not be further defined and explained in subsequent figures.
In the description of the embodiments of the present invention, it should be noted that, if the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. indicate an orientation or a positional relationship based on an orientation or a positional relationship shown in the drawings, or an orientation or a positional relationship which is usually arranged when the product of the present invention is used, it is only for convenience of description and simplification of the description, but does not indicate or imply that the device or the element referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus, should not be construed as limiting the present invention. Furthermore, the terms "first," "second," "third," and the like are used solely to distinguish one from another, and are not to be construed as indicating or implying relative importance.
Furthermore, the terms "horizontal", "vertical", "overhang" and the like do not require that the components be absolutely horizontal or overhang, but may be slightly inclined. For example, "horizontal" merely means that the direction is more horizontal than "vertical" and does not mean that the structure must be perfectly horizontal, but may be slightly inclined.
In the description of the embodiments of the present invention, "a plurality" represents at least 2.
In the description of the embodiments of the present invention, it should be further noted that unless otherwise explicitly stated or limited, the terms "disposed," "mounted," "connected," and "connected" should be broadly construed and interpreted as including, for example, fixed connections, detachable connections, or integral connections; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.
Example 1
Referring to fig. 1 and 2, fig. 1 is a schematic structural diagram of a fluorescence detection card for environmental pollutants according to an embodiment of the present invention, and fig. 2 is a bottom view of fig. 1.
The utility model provides an environmental pollution's fluorescence detection card, including first fluorescence detection board 1, be equipped with three first detection groove 15 on the first fluorescence detection board 1, the bottom that first detection groove 15 was equipped with the first detection window 8 of detachable, one side of first fluorescence detection board 1 articulates there is the second fluorescence detection board 3 that can overlap rather than, and first fluorescence detection board 1 is the same with second fluorescence detection board 3 shape, one side of second fluorescence detection board 3 is equipped with the second that can overlap with first detection groove 15 and detects groove 4, the bottom that the groove 4 was detected to the second is equipped with detachable second and detects window 7, first detection window 8, second detects window 7 and is made by transparent material.
When detecting environmental pollutants with less contrast data and turning the second fluorescence detection plate 3 to the upper side of the first fluorescence detection plate 1, the first detection groove 15 is in butt joint with the second detection groove 4, the second detection window 7 is detached, the first detection groove 15 and the second detection groove 4 are in an open state, when the first detection groove 15 and the second detection groove 4 are longitudinally overlapped, three groups of detection grooves (the first detection groove 15 and the second detection groove 4 are integrated) exist, the floor area of the fluorescence detection card is reduced, the fluorescence detection card is easy to store, meanwhile, the depth of the three groups of detection grooves (the first detection groove 15 and the second detection groove 4 are integrated) is large, and a large amount of reagents and detection samples can be filled for detection. When detecting environmental pollutants with more contrast data, the second fluorescence detection plate 3 is turned away from the upper side of the first fluorescence detection plate 1, the second fluorescence detection plate 3 and the first fluorescence detection plate 1 are arranged in parallel, the first detection window 8 and the second detection window 7 respectively seal one end ports of the first detection groove 15 and the second detection groove 4, detection samples and reagents with different amounts are respectively added into six detection grooves (the three first detection grooves 15 and the three second detection grooves 4), and detection of multiple data can be realized. Above-mentioned two kinds of detection methods (first detect groove 15 and second and detect the switching that groove 4 overlaps and separates), can realize the use of the different detection states of same fluorescence detection card, the application range of fluorescence detection card has been increased, the use limitation of fluorescence detection card has been reduced, and simultaneously, first detection window 8, the connection dismantled of second detection window 7, make first detection groove 15, the both ends that groove 4 was detected to the second all are in open state, be convenient for to first detection groove 15, the second detects groove 4 and washs.
The first fluorescence detection plate 1 and the second fluorescence detection plate 3 are both made of plastics, three first detection grooves 15 are coaxially arranged on the first fluorescence detection plate 1, three second detection grooves 4 are coaxially arranged on the second fluorescence detection plate 3, and the inner sides of the first detection grooves 15 and the second detection grooves 4 are made of transparent materials. The first fluorescence detection plate 1 and the second fluorescence detection plate 3 made of plastics can reduce the overall weight of the invention and have the advantage of portability, the three first detection grooves 15 and the three second detection grooves 4 are coaxially arranged, so that the appearance attractiveness of the invention can be improved, and the inner sides of the first detection grooves 15 and the second detection grooves 4 are made of transparent materials, so that the light reflection efficiency can be improved.
Referring to fig. 1 and 5, fig. 1 is a schematic structural view of a fluorescence detection card for environmental pollutants according to an embodiment of the present invention, fig. 5 is a schematic partial view of a first fluorescence detection plate 1 and a second fluorescence detection plate 3 stacked according to an embodiment of the present invention, the first fluorescence detection plate 1 and the second fluorescence detection plate 3 are strip-shaped, opposite sides of the first fluorescence detection plate 1 and the second fluorescence detection plate 3 are hinged by a hinge 2, and one sides of the first fluorescence detection plate 1 and the second fluorescence detection plate 3 away from the hinged position are connected by a lock catch 13. Hinge 2 can realize the quick articulated between first fluorescence pick-up plate 1 and the second fluorescence detection, and can reduce the gap of first fluorescence pick-up plate 1 and the relative hinged end of second fluorescence pick-up plate 3, increases the compactness between first fluorescence pick-up plate 1 and the second fluorescence pick-up plate 3, and when hasp 13 can realize first fluorescence pick-up plate 1 and the overlap of second fluorescence pick-up plate 3, the fixed connection of the two has avoided rocking relatively between the two.
Referring to fig. 1, 2 and 4, fig. 1 is a schematic structural view of a fluorescence detection card for environmental pollutants according to an embodiment of the present invention, fig. 2 is a bottom view of fig. 1, fig. 4 is a side view of fig. 1, one sides of a first fluorescence detection plate 1 and a second fluorescence detection plate 3 away from a hinge joint are detachably connected by a lock catch 13, an annular groove 5 is disposed in an upper side of the second detection groove 4, and a protrusion abutting against the annular groove 5 is disposed on an upper side of a first detection groove 15 and is made of a transparent material. The first detection groove 15 and the second detection groove 4 are in butt joint with the annular groove 5 through soil blocks, so that the time structure compactness of the first detection groove 15 and the second detection groove 4 can be improved when the first detection groove and the second detection groove are overlapped.
One side of the first detection window 8 is hinged with the first fluorescence detection plate 1, and the other side of the first detection window 8 is detachably connected with the first fluorescence detection plate 1. First detection window 8 is articulated with first fluorescence detection board 1 between, and one side and the second fluorescence detection board 3 of second detection window 7 are articulated, and the opposite side and the second fluorescence detection board 3 of second detection window 7 can be dismantled and be connected, drop from first detection fluorescence detection board, second fluorescence detection board 3 when avoiding first detection window 8, second detection window 7 to dismantle, lead to first detection window 8, second detection window 7 to damage.
Referring to fig. 2 and 3, fig. 2 is a bottom view of fig. 1, fig. 3 is a full-sectional view of fig. 2 a, a clamping groove 11 is disposed on one side of the first fluorescence detection plate 1 facing the first detection window 8, a spherical limiting groove 12 capable of being in coaxial line butt joint with the clamping groove 11 is disposed on one side of the first detection window 8 away from the hinge joint, a spring 10 in a compressed state is disposed in the clamping groove 11, a spherical limiting body 9 capable of being in butt joint with the spherical limiting groove 12 is connected to one side of the spring 10, and the spherical limiting body 9 cannot fall off from the clamping groove 11. The spherical limiting body 9 can be contacted and separated with the spherical limiting groove 12 through the compression spring 10, and the detachable connection between the first detection window 8 and the first fluorescence detection plate 1 is achieved.
One side towards second detection window 7 is equipped with draw-in groove 11 in the second fluorescence detection board 3, and one side that its articulated department was kept away from to second detection window 7 is equipped with spherical spacing groove 12 that can dock with draw-in groove 11 coaxial line, is equipped with the spring 10 that is in compression state in draw-in groove 11, and spring 10 one side is connected with the spherical spacing body 9 that can dock with spherical spacing groove 12, and spherical spacing body 9 can not drop from draw-in groove 11. The spherical limiting body 9 can be contacted and separated with the spherical limiting groove 12 through the compression spring 10, so that the detachable connection between the second detection window 7 and the second fluorescence detection plate 3 is realized.
Referring to fig. 7, fig. 7 is a schematic structural view of the first fluorescence detection plate and the second fluorescence detection plate 3 when not connected to the first detection window and the second detection window according to the embodiment of the present invention, the first detection window 8 and the second detection window 7 are both in a threaded column shape, screw portions of the threaded column-shaped first detection window 8 and the threaded column-shaped second detection window 7 are in a cylindrical shape, and the threaded column-shaped first detection window 8 and the threaded column-shaped second detection window 7 are respectively connected to the first detection groove 15 and the second detection groove 4 in a threaded manner. First detection window 8, second detection window 7 and first detection groove 15, second detection groove 4 pass through threaded connection, have the advantage of high-speed joint and antiseep.
Example 2
Referring to fig. 1 and fig. 6, fig. 1 is a schematic structural diagram of a fluorescence detection card for environmental pollutants according to an embodiment of the present invention, and fig. 6 is a schematic structural diagram of a detection method for detecting environmental pollutants by using a fluorescence detection card according to an embodiment of the present invention, the detection method for detecting environmental pollutants by using a fluorescence detection card:
adjusting: adjusting the first fluorescence detection plate 1 and the second fluorescence detection plate 3 to enable the first fluorescence detection plate 1 and the second fluorescence detection plate 3 to be overlapped, and opening three second detection windows 7 on the second fluorescence detection plate 3;
adding a reagent: filling the three pairs of overlapped first detection grooves 15 and second detection grooves 4 with a testosterone comamonas non-cell system reagent for expressing 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase through three opened second detection windows 7;
adding a detection sample, namely adding environmental pollutants into the overlapped first detection groove 15 and the second detection groove 4, incubating the environmental pollutants with a Comamonas testosteroni non-cell system reagent for expressing 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase at 37 ℃ for 10 minutes, and expressing the Comamonas testosteroni non-cell expression system reagent for expressing 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase to generate green fluorescent protein;
photoelectric treatment: by utilizing the principle of the photoelectric technology, the first fluorescence detection plate 1, the second fluorescence detection plate 3 and the photoelectric technology detection means are organically combined to design an environmental dye detector 14, the fluorescence intensity of expressed green fluorescent protein is detected to judge the concentration of organic pollutants in the environmental pollutants, the degree of the environmental pollutants is evaluated, the 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase expressible testosterone comamonas is replaced by the green fluorescent protein reporter gene through the transgenic technology to construct the 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase fluorescence mutant strain of testosterone comamonas, the mutant strain can efficiently express the green fluorescent protein in the presence of the organic pollutants, through a large amount of enrichment and thallus breakage, various organelles and green fluorescent protein reporter genes in the thallus are obtained, and the transformed Comamonas testosteroni non-cell system reagent of the 3 alpha-hydroxysteroid dehydrogenase/carbonyl acyl reductase is prepared.
After the first fluorescence detection plate 1 and the second fluorescence detection plate 3 are overlapped, three groups of detection grooves (a first detection groove 15 and a second detection groove 4 are integrated) can appear, detection samples and reagents with different amounts can be correspondingly added into the three groups of detection grooves, and then the detection of environmental pollutant samples on a small scale (within three groups) can be realized through photoelectric technology processing.
Example 3
Referring to fig. 1 and fig. 6, fig. 1 is a schematic structural diagram of a fluorescence detection card for environmental pollutants according to an embodiment of the present invention, and fig. 6 is a schematic structural diagram of a detection method for detecting environmental pollutants by using a fluorescence detection card according to an embodiment of the present invention, the detection method for detecting environmental pollutants by using a fluorescence detection card:
adjusting: adjusting the first fluorescence detection plate 1 and the second fluorescence detection plate 3, enabling the second fluorescence detection plate 3 to be turned away from the first fluorescence detection plate 1 and arranged side by side with the first fluorescence detection plate 1, and enabling the first detection window 8 and the second detection window 7 to be respectively in close fit with the bottoms of the first detection groove 15 and the second detection groove 4;
adding a reagent: filling the first detection groove 15 and the second detection groove 4 with a Comamonas testosteroni non-cell system reagent for expressing 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase through the upper openings of the first detection groove 15 and the second detection groove 4 respectively;
adding a detection sample, namely respectively adding environmental pollutants into a first detection groove 15 and a second detection groove 4, incubating the environmental pollutants with a Comamonas testosteroni non-cell system reagent for expressing 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase at 37 ℃ for 10 minutes, and expressing the Comamonas testosteroni non-cell expression system reagent for expressing 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase to generate green fluorescent protein;
photoelectric treatment: by utilizing the principle of the photoelectric technology, the first fluorescence detection plate 1, the second fluorescence detection plate 3 and the photoelectric technology detection means are organically combined to design an environmental pollutant detector 14, the fluorescence intensity of expressed green fluorescent protein is detected to judge the concentration of organic pollutants in the environmental pollutants, the degree of the environmental pollutants is evaluated, the 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase expressible of the 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase gene in the Comamonas testosteroni is replaced by the green fluorescent protein reporter gene through the transgenic technology to construct the 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase fluorescent mutant strain of the Comamonas testosteroni, the mutant strain can efficiently express the green fluorescent protein in the presence of the organic pollutants, through a large amount of enrichment and thallus breakage, various organelles and green fluorescent protein reporter genes in the thallus are obtained, and the transformed Comamonas testosteroni non-cell system reagent of the 3 alpha-hydroxysteroid dehydrogenase/carbonyl acyl reductase is prepared.
After the first fluorescence detection plate 1 is separated from the second fluorescence detection plate 3, three groups of first detection grooves 15 and three groups of second detection grooves 4 can appear, detection samples and reagents with different amounts can be added into the three groups of first detection grooves 15 and the three groups of second detection grooves 4 correspondingly, and then the detection of large-scale (within six groups) environmental pollutant samples can be realized through photoelectric technology processing.
In summary, the invention provides a fluorescence detection card for environmental pollutants, which has at least the following beneficial effects:
when the fluorescence detection card detects environmental pollutants with less contrast data, when the second fluorescence detection plate 3 is turned over to the upper side of the first fluorescence detection plate 1, the first detection groove 15 is in butt joint with the second detection groove 4, the second detection window 7 is detached, the first detection groove 15 and the second detection groove 4 are in an open state, when the first detection groove 15 and the second detection groove 4 are longitudinally overlapped, three groups of detection grooves (the first detection groove 15 and the second detection groove 4 are integrated) exist, the occupied area of the fluorescence detection card is reduced, the fluorescence detection card is easy to store, meanwhile, the depth of the three groups of detection grooves (the first detection groove 15 and the second detection groove 4 are integrated) is large, and a large amount of reagents and detection samples can be filled for detection. When detecting environmental pollutants with more contrast data, the second fluorescence detection plate 3 is turned away from the upper side of the first fluorescence detection plate 1, the second fluorescence detection plate 3 and the first fluorescence detection plate 1 are arranged in parallel, the first detection window 8 and the second detection window 7 respectively seal one end ports of the first detection groove 15 and the second detection groove 4, detection samples and reagents with different amounts are respectively added into six detection grooves (the three first detection grooves 15 and the three second detection grooves 4), and detection of multiple data can be realized. Above-mentioned two kinds of detection methods (first detect groove 15 and second and detect the switching that groove 4 overlaps and separates), can realize the use of the different detection states of same fluorescence detection card, the application range of fluorescence detection card has been increased, the use limitation of fluorescence detection card has been reduced, and simultaneously, first detection window 8, the connection dismantled of second detection window 7, make first detection groove 15, the both ends that groove 4 was detected to the second all are in open state, be convenient for to first detection groove 15, the second detects groove 4 and washs.
After the first fluorescence detection plate 1 and the second fluorescence detection plate 3 are overlapped, three groups of detection grooves (a first detection groove 15 and a second detection groove 4 are integrated) can appear, detection samples and reagents with different amounts can be correspondingly added into the three groups of detection grooves, and then the detection of environmental pollutant samples on a small scale (within three groups) can be realized through photoelectric technology processing.
After the first fluorescence detection plate 1 is separated from the second fluorescence detection plate 3, three groups of first detection grooves 15 and three groups of second detection grooves 4 can appear, detection samples and reagents with different amounts can be added into the three groups of first detection grooves 15 and the three groups of second detection grooves 4 correspondingly, and then the detection of large-scale (within six groups) environmental pollutant samples can be realized through photoelectric technology processing.
The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and various modifications and changes will occur to those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. A detection method for detecting environmental pollutants by adopting a fluorescence detection card, wherein the fluorescence detection card comprises a first fluorescence detection plate, three first detection grooves are arranged on the first fluorescence detection plate, a detachable first detection window is arranged at the bottom of each first detection groove, a second fluorescence detection plate which can be overlapped with the first fluorescence detection plate is hinged to one side of the first fluorescence detection plate, the first fluorescence detection plate and the second fluorescence detection plate are in the same shape, a second detection groove which can be overlapped with the first detection groove is arranged at one side of the second fluorescence detection plate, a detachable second detection window is arranged at the bottom of the second detection groove, and the first detection window and the second detection window are both made of transparent materials, and the method is characterized by comprising the following steps of:
adjusting: adjusting the first fluorescence detection plate and the second fluorescence detection plate to enable the first fluorescence detection plate and the second fluorescence detection plate to be overlapped, and opening three second detection windows on the second fluorescence detection plate;
adding a reagent: filling the three pairs of overlapped first detection grooves and second detection grooves with a testosterone comamonas non-cell system reagent for expressing 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase through three opened second detection windows;
adding a detection sample, namely adding environmental pollutants into the overlapped first detection groove and the second detection groove, incubating the environmental pollutants with a Comamonas testosteroni non-cell system reagent for expressing 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase at 37 ℃ for 10 minutes, and expressing the Comamonas testosteroni non-cell expression system reagent for expressing 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase to generate green fluorescent protein;
photoelectric treatment: by utilizing the principle of a photoelectric technology, the first fluorescence detection plate, the second fluorescence detection plate and a photoelectric technology detection means are organically combined to design an environmental dye detector, the fluorescence intensity of expressed green fluorescent protein is detected to judge the concentration of organic pollutants in the environmental pollutants, the degree of the environmental pollutants is evaluated, the Comamonas testosteroni capable of expressing 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase is replaced by a green fluorescent protein reporter gene through a transgenic technology to construct a Comamonas testosteroni fluorescent variant strain of the 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase, the variant strain can efficiently express the green fluorescent protein in the presence of the organic pollutants, and the strains are increased and crushed, obtaining various organelles in the thallus and green fluorescent protein reporter genes to prepare the transformed Comamonas testosteroni non-cell system reagent of the 3 alpha-hydroxysteroid dehydrogenase/carbonyl reductase.
2. The method as claimed in claim 1, wherein the first and second fluorescence detection plates are made of plastic, three first detection grooves are coaxially formed on the first fluorescence detection plate, three second detection grooves are coaxially formed on the second fluorescence detection plate, and inner sides of the first and second detection grooves are made of transparent material.
3. The method as claimed in claim 2, wherein the first fluorescence detection plate and the second fluorescence detection plate are strip-shaped, opposite sides of the first fluorescence detection plate and the second fluorescence detection plate are hinged by a hinge, and one sides of the first fluorescence detection plate and the second fluorescence detection plate, which are far away from the hinge, are connected by a lock catch.
4. The detecting method for detecting environmental pollutants by using a fluorescence detecting card as claimed in claim 3, wherein the sides of the first fluorescence detecting plate and the second fluorescence detecting plate away from the hinge joint are detachably connected by a lock catch, an annular groove is arranged in the upper side of the second detecting groove, a convex block butted with the annular groove is arranged on the upper side of the first detecting groove, and the convex block is made of a transparent material.
5. A method as recited in claim 1, wherein one side of said first detection window is hingedly connected to said first fluorescence detection plate, said other side of said first detection window is removably connected to said first fluorescence detection plate, said one side of said second detection window is hingedly connected to said second fluorescence detection plate, and said other side of said second detection window is removably connected to said second fluorescence detection plate.
6. The detecting method for detecting environmental pollutants by using a fluorescence detecting card as claimed in claim 5, wherein a slot is formed in one side of the first fluorescence detecting plate facing the first detecting window, a spherical limiting groove capable of being in coaxial line butt joint with the slot is formed in one side of the first detecting window away from the hinge joint of the first detecting window, a spring in a compressed state is arranged in the slot, a spherical limiting body capable of being in butt joint with the spherical limiting groove is connected to one side of the spring, and the spherical limiting body cannot fall off from the slot.
7. The detecting method for detecting environmental pollutants by using the fluorescent detecting card as claimed in claim 5, wherein a slot is provided at one side of the second fluorescent detecting board facing the second detecting window, a spherical limiting groove capable of being in coaxial line butt joint with the slot is provided at one side of the second detecting window away from the hinge joint, a spring in a compressed state is provided in the slot, a spherical limiting body capable of being in butt joint with the spherical limiting groove is connected to one side of the spring, and the spherical limiting body cannot fall off from the slot.
8. The method as claimed in claim 5, wherein the first and second windows are in a threaded cylindrical shape, and the screw portions of the first and second windows are in a cylindrical shape, and the threaded cylindrical windows are respectively connected to the first and second detection grooves by threads.
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