Portable infiltration type detection test paper tube for detecting organophosphorus pesticide

Abstract

The utility model provides a portable infiltration type detection test paper tube for detecting organophosphorus pesticides. The portable infiltration type detection test paper tube comprises a test paper mounting tube, a test paper sheet and filter paper, the test paper mounting barrel is of a barrel-shaped structure with an upper opening, the test paper mounting barrel comprises a bottom barrel with an upper opening and a vertically-through unit barrel section, and the upper end of the bottom barrel is detachably connected with the lower end of the unit barrel section in a sealed butt joint manner; annular supporting plates are fixedly arranged on the inner wall of the bottom cylinder and the inner walls of the unit cylinder sections, test paper sheets are placed on the supporting plates in the bottom cylinder, and filter paper is placed on the supporting plates in the unit cylinder sections; the test paper sheet is provided with a water passing hole which is through up and down, and the test paper sheet comprises a detection area and a contrast area; a UCNPs-Apt layer and an AuNPs-cDNA layer are sequentially arranged in the detection region from bottom to top, and the UCNPs-Apt layer is made of CIT-NaYF4: Yb < 3 + >, Tm < 3 + >-Apt; the control area comprises a UCNPs layer, and the UCNPs layer is made of NaYF4: Yb < 3 + >, Tm < 3 + >.

Description

TECHNICAL FIELD

[0001] The utility model relates to detection test paper technical field especially is portable infiltration type detection test paper tube for detecting organophosphorus pesticides. BACKGROUND

[0002] Agricultural production is the key to maintain food safety and social and economic stability, with the sustained development of modern agriculture, pesticides play an important role in improving crop yield, preventing and treating diseases and pests, reducing production cost and reducing farmers' economic burden. Organophosphorus pesticides (OPs) are phosphoric acid or phosphoric acid derivatives containing phosphorus (P = O) or phosphorus (P = S) thio phosphoryl group, which has the characteristics of high efficiency, broad spectrum, low price, etc. It has been widely used.

[0003] OPs has been widely used due to its wide biological activity, strong applicability, cost-effectiveness and other advantages. Studies have shown that low-dose OPs can cause liver dysfunction, blood toxicity, immune abnormalities, embryotoxicity, genetic toxicity, teratogenicity, developmental neurotoxicity and behavior changes in the body. Therefore, the development of simple, rapid, sensitive and accurate OPs residue detection method has important and far-reaching significance for the safety of living organisms and the protection of ecological environment.

[0004] At present, the main methods for detecting OPs residues in the environment and food are gas chromatography, high performance liquid chromatography, gas chromatography-mass spectrometry, enzyme-linked immunosorbent assay, thin layer chromatography-enzyme inhibition method and spectral analysis method, etc. Although they can achieve trace level detection, sensitivity, reliability and strong specificity, but gas chromatography and gas chromatography-mass spectrometry technology both need large and precise instrument equipment, the detection cost is high, the technical level of the detection personnel is high, and the on-site rapid detection cannot be realized; Enzyme-linked immunosorbent assay, thin layer chromatography-enzyme inhibition method has low detection precision and is easily affected by external factors, resulting in false positive or false negative results; Spectral analysis method is not suitable for trace analysis, has low detection sensitivity and large relative error.

[0005] The current OPs test paper product mainly uses the enzyme inhibition of OPs on acetylcholine esterase. OPs all have enzyme inhibition, this method can only qualitatively detect OPs species, and does not have specific detection function. And the traditional paper-based test paper piece uses capillary action to absorb the sample solution to be measured, and the action time is long. Utility model content

[0006] In order to solve the problem of long action time of traditional paper-based test paper piece using capillary action to absorb the sample solution to be measured in the background art, the utility model provides a portable infiltration type detection test paper tube for detecting organophosphorus pesticides.

[0007] The utility model discloses a portable infiltrating type test paper tube for detecting organic phosphorus pesticide, which comprises a test paper mounting tube, test paper sheets and filter paper.

[0008] The test paper mounting tube is in a cylindrical structure with an upper opening, and comprises a bottom cylinder with an upper opening and a unit cylinder section which is permeable from top to bottom.

[0009] An annular support plate is fixed on the inner wall of the bottom cylinder and the unit cylinder section, and the test paper sheets are placed on the support plate in the bottom cylinder, and the filter paper is placed on the support plate in the unit cylinder section.

[0010] The test paper sheet is provided with a water passing hole which is permeable from top to bottom, and comprises a detection area and a control area.

[0011] The detection area comprises, from bottom to top, a UCNPs-Apt layer and an AuNPs-cDNA layer, and the material of the UCNPs-Apt layer is CIT-NaYF4:Yb 3+ ,Tm 3+ -Apt.

[0012] The control area comprises a UCNPs layer, and the material of the UCNPs layer is NaYF4:Yb 3+ ,Tm 3+ .

[0013] Preferably, the test paper mounting tube is made of transparent material.

[0014] Preferably, the test paper mounting tube comprises a plurality of unit cylinder sections, and the upper and lower ends of two unit cylinder sections are detachably connected in a sealed manner.

[0015] The uppermost support plate is provided with the filter paper, and the other support plates are provided with the test paper sheets.

[0016] The detection areas on the test paper sheets are respectively used for detecting different target Ops.

[0017] The water passing holes on the test paper sheets are arranged in a staggered manner.

[0018] Preferably, a water passing pipe is inserted into the water passing hole, and the upper end of the water passing pipe extends above the upper surface of the test paper sheet.

[0019] Preferably, the upper end of the water passing pipe extends above the upper surface of the test paper sheet by 1mm-3mm.

[0020] Preferably, a pressing sheet is fixed on the inner wall of the bottom cylinder and the unit cylinder section, the vertical section of the pressing sheet is in an L-shaped structure, and the lower end of the pressing sheet is used for pressing on the upper surface of the test paper sheet or the filter paper.

[0021] Preferably, a base is fixed on the bottom of the bottom cylinder, and the diameter of the base is greater than the outer diameter of the bottom cylinder.

[0022] The utility model discloses the advantages are: (1), the test paper piece mixes rare earth doped up-conversion nanometer material as fluorescent material, and this material has long luminescence life, stable chemical property, low toxicity, surface modification, little light damage to biological sample, no spontaneous fluorescence interference, stable fluorescent performance and many advantages, so that the test paper piece preserved according to the regulation has longer shelf life.

[0023] (2), compared with traditional colorimetry and conventional large instrument detection, the equipment has the advantages such as portable, high efficiency and accurate, etc. When carrying out on-site sample detection, the test paper piece can be used to carry out qualitative analysis on actual sample, and simple and accurate screening is carried out on the large batch of sample in the early stage, which is convenient for more accurate analysis and detection of screened sample subsequently.

[0024] (3), compared with traditional enzyme inhibition OPs test paper piece, based on the specific binding principle between aptamer complementary chain, the specific detection of OPs is realized. In addition, different OPs aptamer complementary chain can also be selected according to actual conditions to carry out specific detection on target OPs.

[0025] (4), the test paper piece can be selected according to actual conditions to set up portable fluorescence enzyme label instrument with 980nm excitation light or hitachi fluorescence spectrophotometer f-4600 with solid sample fixing frame to carry out analysis and detection.

[0026] (5), the test paper piece detects the specific numerical value of fluorescence signal, and compared with traditional colorimetry, the detection result is more accurate and sensitive.

[0027] (6), the straight cylinder soaking type sample cell device is adopted, compared with traditional capillary action paper base test paper piece, the reaction area is larger, so that the on-site rapid specific classification qualitative and quantitative detection of OPs is realized. The detection time is shorter.

[0028] (7), the test paper piece design of plug-in type is adopted, and different OPs nucleic acid aptamer and complementary chain test paper piece can be selected according to actual conditions to carry out specific detection on target OPs.

[0029] (8), the to-be-measured solution passes through the upper part of the straight cylinder structure, and most of the large-particle impurities are filtered clean and transparent due to the filtering effect of filter paper. The influence of little impurities on the determination result is reduced, and the accuracy of the test paper piece detection is further improved. BRIEF DESCRIPTION OF DRAWINGS

[0030] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or prior art description. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without creative effort on the basis of these drawings.

[0031] Figure 1 It is a schematic diagram of the main structure of example 1.

[0032] Figure 2 It is Figure 1 The enlarged view of the structure at A in

[0033] Figure 3 It is Figure 1 The schematic diagram of the planar structure of the top angle of the test paper sheet in

[0034] In the figure, 1 is a bottom cylinder, 2 is a base, 3 is a unit cylinder section, 301 is an upper butt joint, 302 is a lower butt joint, 4 is a support plate, 5 is a test paper sheet, 6 is a water pipe, 7 is a detection area, 8 is a control area, 9 is a pressing sheet, and 10 is a filter paper. DETAILED DESCRIPTION

[0035] The technical solutions in the embodiments of the present application will be described clearly and completely below in combination with the drawings in the embodiments of the present application. Obviously, the described embodiments are only some embodiments of the present application, not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative effort fall within the scope of the present application.

[0036] Example 1: A portable infiltrating test paper cylinder for detecting organophosphorus pesticides, comprising a test paper installation cylinder, a test paper sheet 5, and a filter paper 10.

[0037] The test paper installation cylinder is a cylinder structure with an upper opening made of transparent acrylic material. The test paper installation cylinder comprises a bottom cylinder 1 with an upper opening and a plurality of unit cylinder sections 3 which are permeable from top to bottom. The upper end of the bottom cylinder 1 and the lower end of the unit cylinder section 3, and the upper and lower ends of the two unit cylinder sections 3 can be sealed and detachably connected. Specifically, as shown in Figure 1 and Figure 2As shown, both the bottom cylinder 1 and the unit cylinder section 3 are fixedly provided with an upper connector 301. The inner diameter of the upper connector 301 is equal to the inner diameter of the bottom cylinder 1 and the unit cylinder section 3. The outer diameter of the upper connector 301 is smaller than the outer diameter of the bottom cylinder 1 and the unit cylinder section 3. The lower end of the unit cylinder section 3 is fixedly provided with a lower connector 302. The outer diameter of the lower connector 302 is equal to the outer diameter of the unit cylinder section 3. The inner diameter of the lower connector 302 is equal to the outer diameter of the upper connector 301. The lower connector 302 is sleeved on the upper connector 301, and the lower connector 302 and the upper connector 301 are threaded together.

[0038] Annular support plates 4 are fixedly installed on the inner wall of the bottom cylinder 1 and the inner wall of the unit cylinder section 3. Filter paper 10 is placed on the uppermost support plate 4, and test paper pieces 5 are placed on the other support plates 4.

[0039] like Figure 3 As shown, the test strip 5 has water-permeable holes that are open at both the top and bottom, and the water-permeable holes on each test strip 5 are staggered. The test strip 5 includes a detection area 7 and a control area 8. In order to improve the wetting effect of the detection liquid on the test strip 5, such as... Figure 1 and Figure 2 As shown, a water-passing tube 6 is inserted into the water-passing hole, and the upper end of the water-passing tube 6 extends to 2 mm above the upper surface of the test paper 5.

[0040] The detection area 7 includes, from bottom to top, a UCNPs-Apt layer and an AuNPs-cDNA layer. The UCNPs-Apt layer uses CIT-NaYF4:Yb material. 3+ ,Tm 3+ -Apt. In order to enable specific detection of different target Ops, the detection area 7 on each test strip 5 is used to detect different target Ops.

[0041] The control region 8 includes a UCNPs layer, which is made of NaYF4:Yb material. 3+ ,Tm 3+ .

[0042] In order to fix the test strip 5 during testing, such as Figure 1 and Figure 2 As shown, pressure plates 9 are fixedly provided on the inner walls of the bottom cylinder 1 and the unit cylinder section 3. The vertical cross section of the pressure plate 9 is an L-shaped structure. The lower end of the pressure plate 9 is used to press against the upper surface of the test paper sheet 5 or the filter paper 10.

[0043] To increase the contact area between the bottom of the test strip mounting tube and the testing platform or ground, thus improving its stability, such as... Figure 1 As shown, a base 2 is fixedly provided at the bottom of the bottom cylinder 1, and the diameter of the base 2 is larger than the outer diameter of the bottom cylinder 1.

[0044] Working principle: the utility model discloses a hexagonal phase NaYF4:Yb 3+ ,Tm 3+ As fluorescent donor, AuNPs as fluorescent acceptor, two kinds of conjugate combination form CIT-NaYF4:Yb 3+ ,Tm 3+ ,Apt-cDNA-AuNPs complex. Through the principle of base complementary pairing, CIT-NaYF4:Yb 3+ ,Tm 3+ ,Apt and AuNPs-cDNA distance is less than 10nm and the emission spectrum of donor and the absorption spectrum of acceptor overlap, satisfy the fluorescence resonance energy transfer (FRET) condition. In the absence of OPs, CIT-NaYF4:Yb 3+ ,Tm 3+ ,Apt and AuNPs-cDNA form complementary chain, CIT-NaYF4:Yb 3+ ,Tm 3+ ,Apt and AuNPs-cDNA distance shortens, fluorescence quenching. When OPs exist in the system, OPs open CIT-NaYF4:Yb 3+ ,Tm 3+ ,Apt and cDNA-AuNPs between complementary chain, cause fluorescence recovery, through the linear relationship between fluorescence intensity and the OPs concentration in the system quantitative detection is carried out.

[0045] Test paper piece 5 when making, through surface ligand exchange method citric acid (CIT) is modified in NaYF4:Yb 3+ ,Tm 3+ Surface, then through amidation reaction makes CIT-NaYF4:Yb 3+ ,Tm 3+ And OPs-Apt coupling, AuNPs is covalently combined with OPs-cDNA through Au-S bond.

[0046] When the system does not exist OPs, CIT-NaYF4:Yb 3+ ,Tm 3+ ,Apt and AuNPs-cDNA (fluorescent acceptor probe) are combined through the principle of base complementary pairing, and the emission spectrum of donor and the absorption spectrum of acceptor overlap FRET occurs between two, cause fluorescence quenching;When the system exists OPs, OPs-Apt is combined with OPs preferentially, CIT-NaYF4:Yb 3+ ,Tm 3+-Apt and AuNPs-cDNA dissociate, resulting in the recovery of fluorescence, and the intensity of the fluorescence recovery is proportional to the concentration of OPs in the sample. Based on this principle, accurate, sensitive and rapid analysis of the residual concentration of OPs is realized. At the same time, different detectors can be selected for detection according to different application scenarios: portable fluorescence enzyme label instrument with 980nm excitation light and F-4600 Hitachi fluorescence spectrophotometer with solid sample fixing frame are used for analysis and detection.

[0047] Method for use: equal amounts of UCNPs-Apt and UCNPs are coated on the T region and C region of the sample pad respectively, and the immobilized components are dried at 37℃ for 30min. The sample pad is pretreated with blocking solution and dried at 37℃ for 3h for fixation. A certain amount of AuNPs-cDNA is incubated with UCNPs-Apt in the T region to combine and dry and fix, and the preparation of test strip 5 is completed.

[0048] The bottom barrel 1, test strip 5 with different types of OPs aptamer, unit barrel section 3 and filter paper 10 are assembled in the structure as shown in Figure 1 During the assembly process, the orientation of the water passing hole on the upper and lower adjacent test strips 5 is adjusted to make the water passing holes on the upper and lower adjacent test strips 5 misaligned.

[0049] A certain amount of sample to be tested is slowly poured into the upper part of the straight barrel structure, and most of the large particle impurities are filtered clean and transparent due to the filtering action of the filter paper 10. The sample solution to be tested wets the sample pad of the lower test strip 5 in turn due to the action of gravity. When the sample solution to be tested contains target OPs, the OPs preferentially combine with UCNPs-Apt and compete with AuNPs-cDNA. The AuNPs-cDNA that is competed off is free in the flowing solution, and the fluorescence at the T region is recovered due to the absence of AuNPs-cDNA; when the sample solution to be tested does not contain target OPs, the fluorescence at the T region is absorbed by AuNPs-cDNA, resulting in fluorescence quenching. Under the irradiation of 980nm excitation light, the fluorescence values of the T region and the C region of the positive result are basically the same, both have strong fluorescence, while the fluorescence value of the T region of the negative result is significantly weaker than that of the C region.

[0050] Example 2: A portable immersion type detection test paper barrel for detecting organophosphorus pesticides, which is different from example 1 in that the upper end of the water passing pipe 6 extends to 1mm above the upper surface of the test strip 5. The other structures are the same as in example 1.

[0051] Example 3: A portable immersion type detection test paper barrel for detecting organophosphorus pesticides, which is different from example 1 in that the upper end of the water passing pipe 6 extends to 3mm above the upper surface of the test strip 5. The other structures are the same as in example 1.

[0052] It will be apparent to those skilled in the art that the application is not limited to the details of the above-exemplified embodiments and that the application can be implemented in other particular forms without departing from the spirit or essential characteristics of the application. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive, the scope of the application being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. No feature of the application is considered critical unless it is expressly stated in the claims.

Claims

1. A portable dipstick test cartridge for detecting organophosphorus pesticides, characterized by: The test paper installation cylinder, test paper sheet (5), filter paper (10); The test paper installation cylinder is an open-top cylinder structure, and the test paper installation cylinder comprises an open-top bottom cylinder (1) and a top-down permeable unit cylinder section (3). The upper end of the bottom cylinder (1) is in sealing detachable butt joint connection with the lower end of the unit cylinder section (3). The inner wall of the bottom cylinder (1) and the inner wall of the unit cylinder section (3) are both fixedly provided with an annular support plate (4). The test paper sheet (5) is placed on the support plate (4) in the bottom cylinder (1), and the filter paper (10) is placed on the support plate (4) in the unit cylinder section (3). The test paper sheet (5) is provided with a top-down permeable water hole. The test paper sheet (5) comprises a detection area (7) and a control area (8). The UCNPs-Apt layer in the detection zone (7) comprises CIT-NaYF4:Yb, Tm-Apt from bottom to top. 3+ 3+ -Apt.​ The UCNPs layer in the control area (8) is made of NaYF4:Yb 3+ ,Tm 3+ .

2. The portable dipstick test cartridge for detecting organophosphorus pesticides according to claim 1, wherein: The test paper installation cylinder is made of transparent material.

3. The portable dipstick test cartridge for detecting organophosphorus pesticides according to claim 1, wherein: The test paper installation cylinder comprises a plurality of unit cylinder sections (3). The upper and lower ends of two unit cylinder sections (3) can be in sealing detachable butt joint connection. The uppermost support plate (4) is provided with the filter paper (10), and the other support plates (4) are provided with the test paper sheets (5). The detection areas (7) on the test paper sheets (5) are respectively used for detecting different target Ops. The water holes on the test paper sheets (5) are arranged in a staggered manner.

4. The portable dipstick test cartridge for detecting organophosphorus pesticides according to claim 1 or 3, wherein: A water passing pipe (6) is inserted in the water hole. The upper end of the water passing pipe (6) extends above the upper surface of the test paper sheet (5).

5. The portable dipstick test cartridge for detecting organophosphorus pesticides according to claim 4, wherein: The upper end of the water passing pipe (6) extends above the upper surface of the test paper sheet (5) by 1mm-3mm.

6. The portable dipstick test cartridge for detecting organophosphorus pesticides according to claim 3, wherein: The inner walls of the bottom cylinder (1) and the unit cylinder section (3) are both fixedly provided with a pressing piece (9). The vertical section of the pressing piece (9) is in L-shaped structure. The lower end of the pressing piece (9) is used to press on the upper surface of the test paper sheet (5) or the filter paper (10).

7. The portable dipstick test cartridge for detecting organophosphorus pesticides according to claim 1, wherein: The bottom of the bottom cylinder (1) is fixedly provided with a base (2). The diameter of the base (2) is greater than the outer diameter of the bottom cylinder (1).

Images