CN103048449A - Chromatographic detection kit based on aptamer, as well as preparation method and detection method thereof - Google Patents
Chromatographic detection kit based on aptamer, as well as preparation method and detection method thereof Download PDFInfo
- Publication number
- CN103048449A CN103048449A CN2013100127510A CN201310012751A CN103048449A CN 103048449 A CN103048449 A CN 103048449A CN 2013100127510 A CN2013100127510 A CN 2013100127510A CN 201310012751 A CN201310012751 A CN 201310012751A CN 103048449 A CN103048449 A CN 103048449A
- Authority
- CN
- China
- Prior art keywords
- aptamer
- streptavidin
- pad
- detection
- kit
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Abstract
The invention discloses a chromatographic detection kit based on an aptamer, as well as a preparation method and detection method of the kit. The test paper of the kit includes a bottom board, and a sample pad, a bonding pad, a cellulose nitrate film and a water absorption pad, which are adhered on the bottom board and overlap one another; a detection line is arranged on the side of the cellulose nitrate film close to the bonding pad, and a quality control line is arranged on the side of the cellulose nitrate film close to the water absorption pad; an A aptamer labeled by colloidal gold is applied on the bonding pad; a composite containing a B aptamer and streptavidin is applied on the detection line; streptavidin is applied on the quality control line; and both of the A aptamer and the B aptamer are aptamers labeled by biotin and capable of specifically identifying a same test object. The chromatographic detection kit can display the result in five minutes only by directly dropping a diluted sample to be tested into a sample hole, is simple, quick, sensitive, easy to read result, and convenient and simple to prepare, and so on, and has the advantages of simplicity in operation and high specificity without using antibodies and instruments.
Description
Technical field
The present invention relates to a kind of medical testing product and preparation method thereof, relate in particular to a kind of chromatography detection kit based on aptamer and preparation method thereof and detection method.
Background technology
Common immunodetection is that special labelled antibody is fixed in first solid support, after the determinand specific bond, be combined with another antibody again, like this by naked eyes or specific detecting instrument, just can the qualitative detection determinand, thus realize specific immunodiagnosis.
The biomacromolecule that the used specific recognition of immunochromatographic method commonly used detects thing all is antibody, react by the antigen-antibody specific recognition, the immune colloidal solid that is marked on antigen or the antibody makes detection line and nature controlling line show certain color, thereby realizes specific immunodiagnosis.It is quick not and sensitive that but the method detects, and also need to use corresponding instrument and equipment with antibody test, operates not easy.
Summary of the invention
The present invention is intended to overcome the deficiencies in the prior art, a kind of chromatography detection kit based on aptamer and preparation method thereof and detection method.By providing a kind of energy to detect thing by the aptamer specific recognition, on conventional colloidal gold immunochromatographimethod kit basis, make the colloidal gold immunochromatographiassay assay reagent box, by developing the color at detection line and nature controlling line, realize detecting.
For achieving the above object, technical scheme provided by the invention is:
The invention provides a kind of chromatography detection kit based on aptamer, comprise Test paper; Described Test paper comprises base plate, the sample pad, pad, nitrocellulose filter and the adsorptive pads that are bonded on the base plate and overlap successively; Side near pad on the described nitrocellulose membrane is provided with detection line, and the side near adsorptive pads on the nitrocellulose membrane is provided with nature controlling line; Scribble the A aptamer of nanometer particle to mark on the described pad; Scribble the compound of B aptamer and Streptavidin on the described detection line; Scribble Streptavidin on the described nature controlling line; Described A aptamer and B aptamer be can specific recognition same detection thing by biotin labeled aptamer; Wherein, described nano particle is nm of gold, tri-iron tetroxide or latex.
Wherein, the dna sequence dna 5 ' end of described A aptamer is by marking sulfhydryl, and 3 ' end is by biotin labeling; Dna sequence dna 3 ' the end of described B aptamer is by biotin labeling.
The sequence of described A aptamer and B aptamer can design and adjust according to thing to be detected, the dna sequence dna of described A aptamer is shown in SEQ ID NO.1 in an embodiment of the present invention, and the dna sequence dna of described B aptamer is shown in SEQ ID NO.2; The dna sequence dna of described A aptamer is shown in SEQ ID NO.3 in another embodiment of the invention, and the dna sequence dna of described B aptamer is shown in SEQ ID NO.4; The dna sequence dna of described A aptamer is shown in SEQ ID NO.5 in another embodiment of the invention, and the dna sequence dna of described B aptamer is shown in SEQ ID NO.4.
Described base plate is the PVC plate; The material of sample pad is poly-Acetate Film; The material of described pad is glass fibre; Described adsorptive pads is absorbent filter; Described Test paper is packed by the rigid plastic card.
The present invention also provides a kind of method for preparing the mentioned reagent box, comprises the steps:
(1) with nanometer particle to mark A aptamer;
(2) the sample pad treating fluid of sample pad being put into the PBS that contains BSA and tween soaks more than the 1h dry for standby;
(3) with step (1) by the A aptamer of nanometer particle to mark again disperse with the PBS buffer diluent that contains sucrose, BSA and tween in, and be sprayed on the pad dry for standby;
(4) compound of detection line position specking B aptamer and Streptavidin on nitrocellulose membrane is in nature controlling line position specking Streptavidin, dry for standby;
(5) sample pad, pad, nitrocellulose filter and adsorptive pads are overlapped successively and be bonded on the base plate, be cut into test strip, test strip is packed with the rigid plastic card.
Wherein, described step (1) is to be the nanometer particle to mark A aptamer of 10-40nm with diameter; Specking described in the step (4) is to be 0.5-1.5 μ M with B aptamer concentration in the compound of the B of detection line position aptamer and Streptavidin, the solution of preferred 1 μ M and concentration are 0.5-1.5mg/mL, the Streptavidin of preferred 1mg/mL is 1:8-12 by volume, and preferred 1:10 is mixed with and forms; The Streptavidin of described specking in the nature controlling line position is that concentration is 1.5-2.5mg/mL, the Streptavidin dilution of preferred 2mg/mL.
The present invention also provides a kind of detection method based on aptamer:
Described method adopts two aptamers of the same detection thing of identification, on conventional chromatography kit basis, make above-mentioned chromatography detection kit, by realizing detecting without the chromatography of antibody at detection line and nature controlling line colour developing after A aptamer and B aptamer and the determinand specific bond: if nature controlling line does not develop the color, then test result is all invalid; If detection line colour developing and nature controlling line do not develop the color also be considered as invalid; If nature controlling line and detection line all develop the color, then being illustrated in has determinand in the test sample.
The invention will be further described below in conjunction with the detailed manufacturing process of kit and principle:
The concrete manufacturing process of kit of the present invention is as follows:
(1) with diameter is the nanometer particle to mark aptamer 1 of 10-40nm, with sterilized water nano particle is diluted, add again aptamer, centrifugal 15 minutes of 10000rpm, remove supernatant, colloid redissolves in 0.l M PBS damping fluid, and repeatable operation is three times like this, the aptamer 1 that mark is good redissolves and is containing 20mM Na3PO4 at last, in the PBS damping fluid of the 0.0l M pH7.4 of 1%PEG6000;
(2) select poly-Acetate Film as the sample pad material, put it in the sample pad treating fluid of the 0.0l M pH7.4PBS damping fluid that contains 3%BSA, 0.05% polysorbas20 and soak after 1 hour, 37 ℃ of oven dry oven dry in 2 hours, 4 ℃ save backup;
(3) select glass fibre as the pad material, the colloid gold label A aptamer of step (1) preparation is scattered in the 0.0l M PBS damping fluid that contains 5% sucrose, 1%BSA, 0.05% polysorbas20 again, be sprayed on the pad of glass fibre material with some film machine again, 37 degree oven dry 2 hours, 4 ℃ save backup;
(4) point sample on the nitrocellulose filter: be dissolved in the sterilized water with the synthetic B aptamer that is marked with biotin, concentration is 1 μ M, be that the volume ratio of the streptavidin 1:10 of 1mg/ml is mixed with concentration, behind the room temperature reaction one hour, the PBS damping fluid that adds again the long-pending 0.01M pH7.4 of monoploid, then move into the centrifugal ultrafiltration pipe of molecular cut off 30000, centrifugal 15 minutes of 6000rpm, add again and the PBS damping fluid of the 0.01M pH7.4 of above-mentioned equal volume, centrifugal 15 minutes of 6000rpm, repeatable operation is three times like this, obtains being equivalent to the B aptamer streptavidin compound of original volume; Streptavidin is dissolved in the sterilized water, and making concentration is the coated nature controlling line streptavidin dilution of 2mg/ml.Point film machine is put respectively detection line (detection line is positioned at the midline position of whole piece Test paper) and nature controlling line position on nitrocellulose filter with B aptamer streptavidin compound and streptavidin dilution, two lines are separated by three millimeters, then this nitrocellulose filter is put vacuum dryer, dry half an hour, dried 1 hour for 37 ℃, good seal also places room temperature for subsequent use again;
(5) after sticking in order respectively sample pad, pad, nitrocellulose filter and the absorbent filter of handling well on the PVC sheet material, the detector bar that is cut into again four millimeters width is packed in the plastic clip, is assembled into the chromatography detection kit.
During use, sample to be measured is joined the point sample hole, sample to be measured under capillary action along sample pad, pad, nitrocellulose filter is to absorbent filter extreme direction chromatography, in chromatography process, detection thing elder generation in the sample to be measured and the nano particle on the glass fiber conjugate pad-A aptamer compound specific bond also make it to redissolve, potpourri after the redissolution continues chromatography to nitrocellulose filter, the B aptamer generation immune response coated with detection line zone on the nitrocellulose filter, under the effect of certain density detection thing, show the lines of corresponding color; Unnecessary nano particle-A aptamer compound continues forward, when the biotin of mark arrives the nature controlling line zone on the A aptamer, is combined with coated streptavidin, and nano particle generation enrichment shows the color lines of nano particle.If when not having analyte in the sample, nano particle-A aptamer compound and B aptamer can be in the effects of detection line zone, and chromatography reacts by biological streptavidin and continue forward.Therefore detection line is colourless, and the color of the aobvious nano particle of nature controlling line.Nature controlling line be used for to be judged the quality condition of kit: when the detection substrate concentration of sample too high, high salt is perhaps arranged, can cause nano particle-A aptamer all to be enriched in detection line, there is not unnecessary collaurum to continue forward, nature controlling line does not just have the enrichment of nanoparticle interest, be shown as colourlessly, illustrate that the microenvironment that detects has exceeded the scope of application of this detection box, the result is invalid.When aptamer breaks away from or is degraded from nanoparticle surface, or other reason causes the kit quality to change, and all can cause nano particle can not occurring on detection line and the nature controlling line, and detection line and nature controlling line are all colourless, show that testing result is invalid.Therefore, in detection, detection line and nature controlling line be the color of aobvious nano particle simultaneously, just shows the credible of testing result.
The color that the nature controlling line that immunochromatographic method of the present invention shows and p-wire are nano particle.Aptamer also can specific recognition detects thing, two same detection things of energy specific recognition be coated on respectively glass fiber conjugate pad in the colloidal gold immunochromatographimethod and the aptamer on the nitrocellulose membrane.Compared with prior art, the present invention uses conventional chromatography detector bar, realization is without the antibody chromatography: the A aptamer of mark and detect thing in conjunction with after, under capillarity, sample will move forward along nitrocellulose membrane, when moving to when being fixed with detection line on another cellulose nitrate that also can specific recognition detects thing (being coated with B aptamer zone), the detection thing that combines the A aptamer in the sample again with B aptamer generation specific binding, with this, the A aptamer of nanoparticle interest mark, detect thing, another B aptamer that also can specific recognition detects thing forms compound, be the color of visible nano particle, so just realized specific immunochromatographyassay assay.When the present invention prepares test strips, need not to use antibody, do not need other reagent during detection yet, only need directly will hold and get 0.12mL behind the test sample product thin up and splash into well, can go out the result after five minutes, have the characteristics such as simple, quick, sensitive, this detection kit need not to use antibody, do not use instrument and equipment, simple to operate, specificity good, the easy interpretation of result, manufacture convenient and easy.
Description of drawings
Fig. 1 is Test paper structural representation in the kit of the present invention;
Fig. 2 is principle schematic of the present invention, wherein, the detection thing is arranged: expression detects substrate concentration and is higher than minimal detectable concentration, and detection line is the color (being dark color among the figure) of nano particle; Without detecting thing: expression detects substrate concentration and is higher than minimal detectable concentration, detection line position colourless (being blank among the figure);
Fig. 3 is as a result process decision chart of immunochromatographyassay assay of the present invention, and wherein, T represents detection line, and C represents nature controlling line.
Among the figure: 1, base plate; 2, sample pad; 3, pad; 4, nitrocellulose filter; 5, detection line; 6, nature controlling line; 7 adsorptive pads.
Embodiment
The present invention is described further below in conjunction with specific embodiment:
Referring to Fig. 1 to Fig. 3, a kind of colloidal gold immunity chromatography detection kit based on aptamer, comprise Test paper, described Test paper comprises base plate 1, the sample pad 2, pad 3, nitrocellulose filter 4 and the adsorptive pads 7 that are bonded on the base plate 1 and overlap successively; Side near pad 3 on the described nitrocellulose membrane 4 is provided with detection line 5, and the side near adsorptive pads 7 on the nitrocellulose membrane 4 is provided with nature controlling line 6; Scribble the A aptamer of nanometer particle to mark on the described pad 3; Scribble the compound of B aptamer and Streptavidin on the described detection line 5; Scribble Streptavidin on the described nature controlling line 6; Described A aptamer and B aptamer be can specific recognition same detection thing by biotin labeled aptamer; Wherein, described nano particle is collaurum.
Wherein, the dna sequence dna 5 ' end of described A aptamer is by marking sulfhydryl, and 3 ' end is by biotin labeling; Dna sequence dna 3 ' the end of described B aptamer is by biotin labeling; The dna sequence dna of described A aptamer is shown in SEQ ID NO.1, and the dna sequence dna of described B aptamer is shown in SEQ ID NO.2; Described base plate 1 is the PVC plate; The material of sample pad 2 is poly-Acetate Film; The material of described pad 3 is glass fibre; Described adsorptive pads 7 is absorbent filter; Described Test paper is packed by the rigid plastic card.
The method of Preparation Example 1 described kit:
(1) aptamer 1 mark colloid gold particle: centrifugal 15 minutes of the 13 nano-colloid gold solution 10000rpm that will synthesize, the sucking-off supernatant, the sterilized water that adds original volume, twice of repetitive operation, the ratio that adds 0.5 O D aptamer according to every milliliter of collaurum, A aptamer dna sequence dna is: 5 '-SHC6-ATC AGG GCT AAA GAG TGC AGA GTT ACT TAG TTT TTT TTT T-3 ' Biotin(SEQ ID NO.1), this adaptive physical efficiency specific recognition lysozyme, under magnetic agitation, slowly join in the colloidal gold solution, room temperature was placed after 24 hours, add 0.l M PB solution, regulate the PB final concentration to 10mM; Add the PBS damping fluid, NaCl concentration is 0.1M in the regulator solution again, wears out 24 hours under the room temperature; Add at last the PBS damping fluid, the NaCl final concentration is 0.3M in the regulator solution, under the room temperature aging after 24 hours 10000rpm centrifugal, remove supernatant, colloid redissolves in 0.l M PBS damping fluid; Repeatable operation is three times like this, and the aptamer 1 that mark is good redissolves and containing 20mM Na3PO4, in the PBS damping fluid of the 0.0l M pH7.4 of 1%PEG 6000;
(2) select poly-Acetate Film as the sample pad material, after the sample pad treating fluid that poly-Acetate Film is put into the 0.0l M pH7.4PBS damping fluid that contains 3%BSA, 0.05% polysorbas20 soaks 1 hour, 37 ℃ of oven dry oven dry in 2 hours, 4 ℃ save backup;
(3) preparation of pad: during use, the collaurum of step (1) preparation is scattered in the 0.0l M pH7.4PBS damping fluid that contains 5% sucrose, 5%BSA, 1%PEG20000,0.05% polysorbas20 again, be sprayed on the pad of glass fibre material with some film machine again, 37 degree oven dry 2 hours, 4 ℃ save backup;
(4) point sample on the nitrocellulose filter: with the synthetic B aptamer that is marked with biotin, its dna sequence dna is: 5 '-GCA GCT AAG CAG GCG GCT CAC AAA ACC ATT CGCATG CGG CTT TTT TTT TT-3 ' Biotin(SEQ ID NO.2), be dissolved in the sterilized water, concentration is 1 μ M, be that the volume ratio of the streptavidin 1:10 of 1mg/ml is mixed with concentration, behind the room temperature reaction one hour, the PBS damping fluid that adds again the long-pending 0.01M pH7.4 of monoploid, then move into molecular cut off 30000 daltonian centrifugal ultrafiltration pipes, centrifugal 15 minutes of 6000rpm, add again and the PBS damping fluid of the 0.01MpH7.4 of above-mentioned equal volume, centrifugal 15 minutes of 6000rpm, repeatable operation is three times like this, obtains being equivalent to the B aptamer streptavidin compound of original volume; Streptavidin is dissolved in the sterilized water, and making concentration is the coated nature controlling line streptavidin dilution of 2mg/ml.Point film machine is put respectively detection line (detection line is positioned at the midline position of whole piece Test paper) and nature controlling line position on nitrocellulose filter with aptamer 2 streptavidin compounds and streptavidin dilution, two lines are separated by three millimeters, then this nitrocellulose filter is put vacuum dryer, dry half an hour, dried 1 hour for 37 ℃, good seal also places room temperature for subsequent use again;
(5) after sticking in order respectively sample pad, pad, nitrocellulose filter and the absorbent filter of handling well on the PVC sheet material, the detector bar that is cut into again four millimeters width is packed in the plastic clip, is assembled into the chromatography detection kit;
(6) detect: draw and get 0.12mL behind the sample thin up to be measured and splash into well.If nature controlling line does not develop the color, test result is all invalid, if the aobvious redness of detection line and nature controlling line do not develop the color also be considered as invalid; If redness all appears in nature controlling line and detection line, being illustrated in has the lysozyme that is higher than minimal detectable concentration (1 μ M) in the test sample.
Embodiment 3
Referring to Fig. 1 to Fig. 3, a kind of colloidal gold immunity chromatography detection kit based on aptamer, comprise Test paper, described Test paper comprises base plate 1, the sample pad 2, pad 3, nitrocellulose filter 4 and the adsorptive pads 7 that are bonded on the base plate 1 and overlap successively; Side near pad 3 on the described nitrocellulose membrane 4 is provided with detection line 5, and the side near adsorptive pads 7 on the nitrocellulose membrane 4 is provided with nature controlling line 6; Scribble the A aptamer of nanometer particle to mark on the described pad 3; Scribble the compound of B aptamer and Streptavidin on the described detection line 5; Scribble Streptavidin on the described nature controlling line 6; Described A aptamer and B aptamer be can specific recognition same detection thing by biotin labeled aptamer; Wherein, described nano particle is collaurum.
Wherein, the dna sequence dna 5 ' end of described A aptamer is by marking sulfhydryl, and 3 ' end is by biotin labeling; Dna sequence dna 3 ' the end of described B aptamer is by biotin labeling; The dna sequence dna of described A aptamer is shown in SEQ ID NO.3, and the dna sequence dna of described B aptamer is shown in SEQ ID NO.4; Described base plate 1 is the PVC plate; The material of sample pad 2 is poly-Acetate Film; The material of described pad 3 is glass fibre; Described adsorptive pads 7 is absorbent filter; Described Test paper is packed by the rigid plastic card.
The method of Preparation Example 3 described kits:
(1) aptamer 1 mark colloid gold particle: centrifugal 15 minutes of the 10 nano-colloid gold solution 10000rpm that will synthesize, the sucking-off supernatant, the sterilized water that adds original volume, twice of repetitive operation, the ratio that adds 0.5OD aptamer 1 according to every milliliter of collaurum, the A aptamer dna sequence dna of specific recognition human thrombin is: 5 ' SH C6-TTT TTT TTT TGG TTG GTG TGG TTG GTT TTTTTT TT3 ' Biotin(SEQ ID NO.3), under magnetic agitation, slowly join in the colloidal gold solution, room temperature was placed after 24 hours, add 100mM PB solution, regulate final concentration to 10mM; Add the PBS damping fluid, NaCl concentration is 0.1M in the regulator solution again, wears out 24 hours under the room temperature; Add at last the PBS damping fluid, the NaCl final concentration is 0.3M in the regulator solution, under the room temperature aging after 24 hours 10000rpm centrifugal, remove supernatant, colloid redissolves in 0.l M PBS damping fluid, repeatable operation is three times like this, and the aptamer 1 that mark is good redissolves and containing 20mM Na3PO4 at last, in the PBS damping fluid of the 0.0l M pH7.4 of 1%PEG6000;
(2) select poly-Acetate Film as the sample pad material, after the sample pad treating fluid that poly-Acetate Film is put into the 0.0l M pH7.4PBS damping fluid that contains 3%BSA, 0.05% polysorbas20 soaks 1 hour, 37 ℃ of oven dry oven dry in 2 hours, 4 ℃ save backup;
(3) preparation of pad: during use, the collaurum of step (1) preparation is scattered in the 0.0l M pH7.2PBS damping fluid that contains 5% sucrose, 2%BSA, 2%Dextran, 0.05% polysorbas20 again, be sprayed on the glass fiber conjugate pad with some film machine again, dried 2 hours for 37 ℃, 4 ℃ save backup;
(4) point sample on the nitrocellulose filter: the specific recognition human thrombin B aptamer that is marked with biotin with a synthetic end, its dna sequence dna is: 5 '-AGT CCG TGG TAG GGC AGG TTGGGG TGA CTT TTT TTT TTT-3 ' Biotin(SEQ ID NO.4), be dissolved in the sterilized water, concentration is 1 μ M, be that the streptavidin of 1mg/ml mixes with the volume ratio of 1:10 with concentration, room temperature reaction one hour, at the PBS damping fluid that adds the long-pending 0.02M pH7.2 of monoploid, then move into molecular cut off 30000 daltonian centrifugal ultrafiltration pipes, centrifugal 15 minutes of 6000rpm, add again and the PBS damping fluid of the 0.02M pH7.2 of above-mentioned equal volume, centrifugal 15 minutes of 6000rpm, repeatable operation is three times like this, obtains being equivalent to the aptamer 2 streptavidin compounds of original volume; Streptavidin is dissolved in the sterilized water, and coated nature controlling line is 2mg/ml with the streptavidin diluent concentration.Point film machine is put respectively detection line (detection line is positioned at whole test strip midline position) and nature controlling line position on nitrocellulose filter with aptamer 2 and streptavidin compound and streptavidin dilution, two lines are separated by three millimeters, then nitrocellulose filter is placed 37 ℃ of oven dry 2 hours, good seal also places room temperature for subsequent use;
(5) after sticking in order respectively sample pad, pad, nitrocellulose filter and the absorbent filter of handling well on the PVC sheet material, the detector bar that is cut into again four millimeters width is packed in the plastic clip, is assembled into the chromatography detection kit;
(6) detect: draw and get 0.12mL behind the sample thin up to be measured and splash into well.If nature controlling line does not develop the color, test result is all invalid, if the aobvious redness of detection line and nature controlling line do not develop the color also be considered as invalid; If redness all appears in nature controlling line and detection line, being illustrated in has the lysozyme that is higher than minimal detectable concentration (1 μ M) in the test sample.
Referring to Fig. 1 to Fig. 3, a kind of colloidal gold immunity chromatography detection kit based on aptamer, comprise Test paper, described Test paper comprises base plate 1, the sample pad 2, pad 3, nitrocellulose filter 4 and the adsorptive pads 7 that are bonded on the base plate 1 and overlap successively; Side near pad 3 on the described nitrocellulose membrane 4 is provided with detection line 5, and the side near adsorptive pads 7 on the nitrocellulose membrane 4 is provided with nature controlling line 6; Scribble the A aptamer of nanometer particle to mark on the described pad 3; Scribble the compound of B aptamer and Streptavidin on the described detection line 5; Scribble Streptavidin on the described nature controlling line 6; Described A aptamer and B aptamer be can specific recognition same detection thing by biotin labeled aptamer; Wherein, described nano particle is ferroferric oxide nano granules.
Wherein, the dna sequence dna 5 ' end of described A aptamer is by marking sulfhydryl; Dna sequence dna 3 ' the end of described B aptamer is by biotin labeling; The dna sequence dna of described A aptamer is shown in SEQ ID NO.5, and the dna sequence dna of described B aptamer is shown in SEQ ID NO.4; Described base plate 1 is the PVC plate; The material of sample pad 2 is poly-Acetate Film; The material of described pad 3 is glass fibre; Described adsorptive pads 7 is absorbent filter; Described Test paper is packed by the rigid plastic card.
Embodiment 6
The method of Preparation Example 5 described kits:
(1) aptamer 1 mark ferroferric oxide nano granules: with the surface with centrifugal 10 minutes of the brown ferroferric oxide nano granules solution 14000rpm of particle diameter 10 nanometers of carboxyl, the sucking-off supernatant, adding concentration is in the MES solution of 10mM EDC, stirring at normal temperature 30 minutes, the centrifugal supernatant that goes, according to the ratio of every milliliter of nanoparticles solution adding 0.5OD aptamer 1, the A aptamer dna sequence dna of the specific recognition human thrombin of the PBS damping fluid dissolving of adding 10mM pH7.4 is: 5 '-TGG TTGGTG TGG TTG GTT TTT TTT TT-3 ' NH
2(SEQ ID NO.5), under magnetic agitation, slowly join in the ferroferric oxide nano granules, room temperature was placed after 2 hours, the centrifugal supernatant that goes, use again the PBS damping fluid washed twice of 10mM pH7.4, the aptamer 1 that mark is good redissolves and is containing 20mM Na3PO4 at last, in the PBS damping fluid of the 0.0l M pH7.4 of 1%PEG6000;
(2) select poly-Acetate Film as the sample pad material, after the sample pad treating fluid that poly-Acetate Film is put into the 0.0l M pH7.4PBS damping fluid that contains 3%BSA, 0.05% polysorbas20 soaks 1 hour, 37 ℃ of oven dry oven dry in 2 hours, 4 ℃ save backup;
(3) preparation of pad: during use, the ferroferric oxide nano granules of step (1) preparation is scattered in the 0.0l M pH7.2PBS damping fluid that contains 5% sucrose, 2%BSA, 2%Dextran, 0.05% polysorbas20 again, be sprayed on the glass fiber conjugate pad with some film machine again, dried 2 hours for 37 ℃, 4 ℃ save backup;
(4) point sample on the nitrocellulose filter: the specific recognition human thrombin B aptamer that is marked with biotin with a synthetic end, its dna sequence dna is: 5 '-AGT CCG TGG TAG GGC AGG TTGGGG TGA CTT TTT TTT TTT-3 ' Biotin(SEQ ID NO.4), be dissolved in the sterilized water, concentration is 1 μ M, be that the streptavidin of 1mg/ml mixes with the volume ratio of 1:10 with concentration, room temperature reaction one hour, at the PBS damping fluid that adds the long-pending 0.02M pH7.2 of monoploid, then move into molecular cut off 30000 daltonian centrifugal ultrafiltration pipes, centrifugal 15 minutes of 6000rpm, add again and the PBS damping fluid of the 0.02M pH7.2 of above-mentioned equal volume, centrifugal 15 minutes of 6000rpm, repeatable operation is three times like this, obtains being equivalent to the aptamer 2 streptavidin compounds of original volume; Streptavidin is dissolved in the sterilized water, and coated nature controlling line is 2mg/ml with the streptavidin diluent concentration.Point film machine is put respectively detection line (detection line is positioned at whole test strip midline position) and nature controlling line position on nitrocellulose filter with aptamer 2 and streptavidin compound and streptavidin dilution, two lines are separated by three millimeters, then nitrocellulose filter is placed 37 ℃ of oven dry 2 hours, good seal also places room temperature for subsequent use;
(5) after sticking in order respectively sample pad, pad, nitrocellulose filter and the absorbent filter of handling well on the PVC sheet material, the detector bar that is cut into again four millimeters width is packed in the plastic clip, is assembled into the chromatography detection kit;
(6) detect: after drawing human blood to be measured and resetting and adding the sterilized water dilution, get 0.12mL and splash into well, if nature controlling line does not develop the color for invalid, if the aobvious brown of detection line and nature controlling line do not develop the color also be considered as invalid, if brown all appears in nature controlling line and detection line, being illustrated in has the human thrombin that is higher than minimal detectable concentration (100nM) in the test sample.
Sequence table
Claims (10)
1. the chromatography detection kit based on aptamer comprises Test paper; Described Test paper comprises base plate (1), is bonded in base plate (1) sample pad (2), pad (3), nitrocellulose filter (4) and adsorptive pads (7) upper and successively overlap joint; The upper side near pad (3) of described nitrocellulose membrane (4) is provided with detection line (5), and the upper side near adsorptive pads (7) of nitrocellulose membrane (4) is provided with nature controlling line (6); It is characterized in that, scribble the A aptamer of nanometer particle to mark on the described pad (3); Scribble the compound of B aptamer and Streptavidin on the described detection line (5); Described nature controlling line scribbles Streptavidin on (6); Described A aptamer and B aptamer be can specific recognition same detection thing by biotin labeled aptamer; Wherein, described nano particle is nm of gold, tri-iron tetroxide or latex.
2. kit as claimed in claim 1 is characterized in that, the dna sequence dna 5 ' end of described A aptamer is by marking sulfhydryl, and 3 ' end is by biotin labeling; Dna sequence dna 3 ' the end of described B aptamer is by biotin labeling.
3. kit as claimed in claim 1 is characterized in that, the dna sequence dna of described A aptamer is shown in SEQ ID NO.1, and the dna sequence dna of described B aptamer is shown in SEQ ID NO.2.
4. kit as claimed in claim 1 is characterized in that, the dna sequence dna of described A aptamer is shown in SEQ ID NO.3, and the dna sequence dna of described B aptamer is shown in SEQ ID NO.4.
5. kit as claimed in claim 1 is characterized in that, the dna sequence dna of described A aptamer is shown in SEQ ID NO.5, and the dna sequence dna of described B aptamer is shown in SEQ ID NO.4.
6. kit as claimed in claim 1 is characterized in that, described base plate (1) is the PVC plate; The material of sample pad (2) is poly-Acetate Film; The material of described pad (3) is glass fibre; Described adsorptive pads (7) is absorbent filter; Described Test paper is packed by the rigid plastic card.
7. prepare the method for each described kit of claim 1 to 6, comprise the steps:
(1) with nanometer particle to mark A aptamer;
(2) the sample pad treating fluid of sample pad being put into the PBS that contains BSA and tween soaks more than the 1h dry for standby;
(3) with step (1) by the A aptamer of nanoparticle interest mark again disperse with the PBS buffer diluent that contains sucrose, BSA and tween in, and be sprayed on the pad dry for standby;
(4) compound of detection line position specking B aptamer and Streptavidin on nitrocellulose membrane is in nature controlling line position specking Streptavidin, dry for standby;
(5) sample pad, pad, nitrocellulose filter and adsorptive pads are overlapped successively and be bonded on the base plate, be cut into test strip, test strip is packed with the rigid plastic card.
8. method as claimed in claim 7 is characterized in that, described step (1) is to be the nanometer particle to mark A aptamer of 10-40nm with diameter; Specking described in the step (4) in the compound of the B of detection line position aptamer and Streptavidin be with B aptamer concentration be the solution of 0.5-1.5 μ M and Streptavidin that concentration is 0.5-1.5mg/mL by volume 1:8-12 be mixed with and form; The Streptavidin of described specking in the nature controlling line position is that concentration is the Streptavidin dilution of 1.5-2.5mg/mL.
9. method as claimed in claim 8, it is characterized in that, specking described in the step (4) in the compound of the B of detection line position aptamer and Streptavidin be with B aptamer concentration be the solution of 1 μ M and Streptavidin that concentration is 1mg/mL by volume 1:10 be mixed with and form; The Streptavidin of described specking in the nature controlling line position is that concentration is the Streptavidin dilution of 2mg/mL.
10. detection method based on aptamer, it is characterized in that, described method adopts two aptamers of the same detection thing of identification, on conventional chromatography kit basis, make each described chromatography detection kit of claim 1 to 6, by realizing detecting without the chromatography of antibody at detection line and nature controlling line colour developing after A aptamer and B aptamer and the determinand specific bond: if nature controlling line does not develop the color, then test result is all invalid; If detection line colour developing and nature controlling line do not develop the color also be considered as invalid; If nature controlling line and detection line all develop the color, then being illustrated in has determinand in the test sample.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310012751.0A CN103048449B (en) | 2013-01-14 | 2013-01-14 | Chromatographic detection kit based on aptamer, as well as preparation method and detection method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310012751.0A CN103048449B (en) | 2013-01-14 | 2013-01-14 | Chromatographic detection kit based on aptamer, as well as preparation method and detection method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103048449A true CN103048449A (en) | 2013-04-17 |
| CN103048449B CN103048449B (en) | 2015-06-24 |
Family
ID=48061163
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310012751.0A Active CN103048449B (en) | 2013-01-14 | 2013-01-14 | Chromatographic detection kit based on aptamer, as well as preparation method and detection method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103048449B (en) |
Cited By (27)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103983774A (en) * | 2014-06-03 | 2014-08-13 | 合肥工业大学 | Probe capable of specifically identifying bisphenol A nucleic acid aptamer and test strip detection application of probe |
| CN104593372A (en) * | 2015-02-09 | 2015-05-06 | 湖南大学 | Aptamer, kit and method for detecting pancreatic duct adenocarcinoma |
| CN104606919A (en) * | 2014-12-31 | 2015-05-13 | 华南师范大学 | Preparation method of novel nucleic acid aptamer/nano-gold modified solid phase microextraction quartz capillary tube |
| CN104745586A (en) * | 2013-12-27 | 2015-07-01 | 上海市刑事科学技术研究院 | Cocaine aptamer, detection kit and application thereof |
| CN105572393A (en) * | 2016-01-21 | 2016-05-11 | 武汉慧禹信息科技有限公司 | Nucleic acid aptamer based on estradiol in saliva and gold mark test strip for detection |
| CN105784990A (en) * | 2016-05-17 | 2016-07-20 | 中国农业科学院农业质量标准与检测技术研究所 | Test strip for detecting aflatoxin B1 or M1 by utilizing aptamer |
| CN106290831A (en) * | 2016-08-02 | 2017-01-04 | 中国农业科学院农业质量标准与检测技术研究所 | A kind of competition law test strip based on antigen aptamers and application thereof |
| CN106568951A (en) * | 2016-10-26 | 2017-04-19 | 北京农业质量标准与检测技术研究中心 | Nucleic acid aptamer-based escherichia coli O157:H7 colloidal gold test strip, and detection method |
| CN106970076A (en) * | 2017-04-17 | 2017-07-21 | 西安交通大学 | It is a kind of to be used for sxemiquantitative aptamer test strips that TNF α are detected and preparation method thereof |
| CN107164383A (en) * | 2016-09-29 | 2017-09-15 | 北京理工大学 | One group of creatine kinase isozyme aptamer and its application |
| CN107462714A (en) * | 2017-07-17 | 2017-12-12 | 昆明理工大学 | With the aptamer Sf A09 of sodium formaldehyde sulfoxylate specific bond and its application |
| CN107478830A (en) * | 2017-06-14 | 2017-12-15 | 江苏大学 | A kind of ochratoxin A Rapid detection test strip based on aptamers identification |
| CN108303415A (en) * | 2018-01-18 | 2018-07-20 | 北京勤邦生物技术有限公司 | A kind of aptamer test strips and its preparation method and application of detection aflatoxin B1 |
| JP2018519796A (en) * | 2015-04-29 | 2018-07-26 | ドッツ テクノロジー コーポレイションＤＯＴＳ Ｔｅｃｈｎｏｌｏｇｙ Ｃｏｒｐ． | Compositions and methods for allergen detection |
| CN109187970A (en) * | 2018-12-05 | 2019-01-11 | 鲁东大学 | It is a kind of quickly to detect aquatic products disease gold mark nucleic acid test strip and preparation method thereof |
| CN109321577A (en) * | 2018-10-09 | 2019-02-12 | 安徽科技学院 | It is a kind of to detect the aptamers group of excretion body, lateral flow type aptamers biosensor and preparation method thereof |
| CN109596827A (en) * | 2019-01-17 | 2019-04-09 | 长江师范学院 | Fluorescence detection test strip and its preparation method and application that is a kind of while detecting 4 kinds of pathogenic bacteria |
| CN109738624A (en) * | 2018-12-29 | 2019-05-10 | 中国科学院广州生物医药与健康研究院 | A kind of colloidal gold aptamer test strips of platelet derived growth factor PDGF-BB and preparation method thereof |
| CN109765381A (en) * | 2018-12-29 | 2019-05-17 | 长沙科雅生物科技有限公司 | A kind of platelet derived growth factor PDGF-BB test strips and detection method based on the amplification of aptamer signal |
| CN109781994A (en) * | 2018-12-29 | 2019-05-21 | 长沙科雅生物科技有限公司 | A kind of platelet derived growth factor PDGF-BB test strips and detection method based on the amplification of aptamer signal |
| CN109900905A (en) * | 2019-02-28 | 2019-06-18 | 中国科学院广州生物医药与健康研究院 | A kind of colloidal gold strip and preparation method detecting carcinomebryonic antigen |
| CN110441508A (en) * | 2019-05-06 | 2019-11-12 | 华南理工大学 | Acrylamide in Foods Rapid detection test strip based on nanogold composite coating |
| CN111044729A (en) * | 2019-12-13 | 2020-04-21 | 西安医学院 | Aptamer-based high-sensitivity C-reactive protein nano immunochromatography detection method |
| CN112114134A (en) * | 2019-06-19 | 2020-12-22 | 爱科来株式会社 | Target substance detection method, target substance detection kit, and target substance detection system |
| CN113174447A (en) * | 2021-03-26 | 2021-07-27 | 江苏省农业科学院 | Gold-labeled rapid detection card for detecting CGMMV and preparation method thereof |
| CN114441521A (en) * | 2022-01-27 | 2022-05-06 | 宜宾五粮液股份有限公司 | Preparation method of white spirit ethyl carbamate quick-detection test paper |
| CN116008542A (en) * | 2022-10-21 | 2023-04-25 | 青岛农业大学 | Aptamer test strip, preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101851667A (en) * | 2009-04-03 | 2010-10-06 | 上海市公共卫生临床中心 | Fluorescent quantitative PCR detection kit of vibrio parahemolyticus pathogenicity marker gene tdh as well as detection method and application thereof |
| CN101942386A (en) * | 2009-07-08 | 2011-01-12 | 中国科学院广州生物医药与健康研究院 | Nucleic acid nanogold biosensor and preparation method thereof |
| CN101942387A (en) * | 2009-07-08 | 2011-01-12 | 中国科学院广州生物医药与健康研究院 | Nucleic acid nano-gold biosensor for detecting influenza A viruses and influenza A (H1N1) viruses |
-
2013
- 2013-01-14 CN CN201310012751.0A patent/CN103048449B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101851667A (en) * | 2009-04-03 | 2010-10-06 | 上海市公共卫生临床中心 | Fluorescent quantitative PCR detection kit of vibrio parahemolyticus pathogenicity marker gene tdh as well as detection method and application thereof |
| CN101942386A (en) * | 2009-07-08 | 2011-01-12 | 中国科学院广州生物医药与健康研究院 | Nucleic acid nanogold biosensor and preparation method thereof |
| CN101942387A (en) * | 2009-07-08 | 2011-01-12 | 中国科学院广州生物医药与健康研究院 | Nucleic acid nano-gold biosensor for detecting influenza A viruses and influenza A (H1N1) viruses |
Non-Patent Citations (3)
| Title |
|---|
| HUI XU ET AL.: "Aptamer-Functionalized Gold Nanoparticles as Probes in a Dry-Reagent Strip Biosensor for Protein Analysis", 《ANALYTICAL CHEMISTRY》, vol. 81, no. 2, 12 December 2008 (2008-12-12), pages 669 - 675 * |
| XUN MAO ET AL: "Disposable nucleic acid biosensors based on gold nanoparticle probes and lateral strip", 《ANALYTICAL CHEMISTRY》, vol. 81, no. 4, 21 January 2009 (2009-01-21), pages 1660 - 1668 * |
| 仲志鸿 等: "以分子信标为报告分子的凝血酶检测新方法", 《生命科学研究》, vol. 10, no. 3, 30 September 2006 (2006-09-30), pages 206 - 209 * |
Cited By (40)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104745586A (en) * | 2013-12-27 | 2015-07-01 | 上海市刑事科学技术研究院 | Cocaine aptamer, detection kit and application thereof |
| CN104745586B (en) * | 2013-12-27 | 2020-02-04 | 上海市刑事科学技术研究院 | Cocaine aptamer, detection kit and application thereof |
| CN103983774A (en) * | 2014-06-03 | 2014-08-13 | 合肥工业大学 | Probe capable of specifically identifying bisphenol A nucleic acid aptamer and test strip detection application of probe |
| CN104606919A (en) * | 2014-12-31 | 2015-05-13 | 华南师范大学 | Preparation method of novel nucleic acid aptamer/nano-gold modified solid phase microextraction quartz capillary tube |
| CN104593372A (en) * | 2015-02-09 | 2015-05-06 | 湖南大学 | Aptamer, kit and method for detecting pancreatic duct adenocarcinoma |
| JP2018519796A (en) * | 2015-04-29 | 2018-07-26 | ドッツ テクノロジー コーポレイションＤＯＴＳ Ｔｅｃｈｎｏｌｏｇｙ Ｃｏｒｐ． | Compositions and methods for allergen detection |
| US10466246B2 (en) | 2015-04-29 | 2019-11-05 | Dots Technology Corp. | Compositions and methods for allergen detection |
| US11016094B2 (en) | 2015-04-29 | 2021-05-25 | Dots Technology Corp. | Compositions and methods for allergen detection |
| CN105572393B (en) * | 2016-01-21 | 2017-11-17 | 武汉慧禹信息科技有限公司 | The aptamer and detection gold label test strip of a kind of estradiol based in saliva |
| CN105572393A (en) * | 2016-01-21 | 2016-05-11 | 武汉慧禹信息科技有限公司 | Nucleic acid aptamer based on estradiol in saliva and gold mark test strip for detection |
| CN105784990B (en) * | 2016-05-17 | 2018-06-01 | 中国农业科学院农业质量标准与检测技术研究所 | It is a kind of to detect aflatoxin B1 or the test strips of M1 using aptamers |
| CN105784990A (en) * | 2016-05-17 | 2016-07-20 | 中国农业科学院农业质量标准与检测技术研究所 | Test strip for detecting aflatoxin B1 or M1 by utilizing aptamer |
| CN106290831A (en) * | 2016-08-02 | 2017-01-04 | 中国农业科学院农业质量标准与检测技术研究所 | A kind of competition law test strip based on antigen aptamers and application thereof |
| CN107164383A (en) * | 2016-09-29 | 2017-09-15 | 北京理工大学 | One group of creatine kinase isozyme aptamer and its application |
| CN106568951B (en) * | 2016-10-26 | 2019-01-01 | 北京农业质量标准与检测技术研究中心 | Escherichia coli O 157 based on aptamer: H7 colloidal gold strip and detection method |
| CN106568951A (en) * | 2016-10-26 | 2017-04-19 | 北京农业质量标准与检测技术研究中心 | Nucleic acid aptamer-based escherichia coli O157:H7 colloidal gold test strip, and detection method |
| CN106970076A (en) * | 2017-04-17 | 2017-07-21 | 西安交通大学 | It is a kind of to be used for sxemiquantitative aptamer test strips that TNF α are detected and preparation method thereof |
| CN107478830A (en) * | 2017-06-14 | 2017-12-15 | 江苏大学 | A kind of ochratoxin A Rapid detection test strip based on aptamers identification |
| CN107462714B (en) * | 2017-07-17 | 2019-06-11 | 昆明理工大学 | With the aptamer Sf-A09 of sodium formaldehyde sulfoxylate specific bond and its application |
| CN107462714A (en) * | 2017-07-17 | 2017-12-12 | 昆明理工大学 | With the aptamer Sf A09 of sodium formaldehyde sulfoxylate specific bond and its application |
| CN108303415A (en) * | 2018-01-18 | 2018-07-20 | 北京勤邦生物技术有限公司 | A kind of aptamer test strips and its preparation method and application of detection aflatoxin B1 |
| CN109321577B (en) * | 2018-10-09 | 2020-12-08 | 安徽科技学院 | Aptamer group for detecting exosome, lateral flow aptamer biosensor and preparation method thereof |
| CN109321577A (en) * | 2018-10-09 | 2019-02-12 | 安徽科技学院 | It is a kind of to detect the aptamers group of excretion body, lateral flow type aptamers biosensor and preparation method thereof |
| CN109187970A (en) * | 2018-12-05 | 2019-01-11 | 鲁东大学 | It is a kind of quickly to detect aquatic products disease gold mark nucleic acid test strip and preparation method thereof |
| CN109781994B (en) * | 2018-12-29 | 2021-12-03 | 长沙科雅生物科技有限公司 | Aptamer signal amplification-based platelet-derived growth factor PDGF-BB test strip and detection method |
| CN109781994A (en) * | 2018-12-29 | 2019-05-21 | 长沙科雅生物科技有限公司 | A kind of platelet derived growth factor PDGF-BB test strips and detection method based on the amplification of aptamer signal |
| CN109765381B (en) * | 2018-12-29 | 2021-12-03 | 长沙科雅生物科技有限公司 | Aptamer signal amplification-based platelet-derived growth factor PDGF-BB test strip and detection method |
| CN109738624A (en) * | 2018-12-29 | 2019-05-10 | 中国科学院广州生物医药与健康研究院 | A kind of colloidal gold aptamer test strips of platelet derived growth factor PDGF-BB and preparation method thereof |
| CN109765381A (en) * | 2018-12-29 | 2019-05-17 | 长沙科雅生物科技有限公司 | A kind of platelet derived growth factor PDGF-BB test strips and detection method based on the amplification of aptamer signal |
| CN109596827A (en) * | 2019-01-17 | 2019-04-09 | 长江师范学院 | Fluorescence detection test strip and its preparation method and application that is a kind of while detecting 4 kinds of pathogenic bacteria |
| CN109900905A (en) * | 2019-02-28 | 2019-06-18 | 中国科学院广州生物医药与健康研究院 | A kind of colloidal gold strip and preparation method detecting carcinomebryonic antigen |
| CN110441508A (en) * | 2019-05-06 | 2019-11-12 | 华南理工大学 | Acrylamide in Foods Rapid detection test strip based on nanogold composite coating |
| CN110441508B (en) * | 2019-05-06 | 2020-11-24 | 华南理工大学 | Test strip for rapidly detecting acrylamide in food based on nanogold composite coating |
| CN112114134A (en) * | 2019-06-19 | 2020-12-22 | 爱科来株式会社 | Target substance detection method, target substance detection kit, and target substance detection system |
| CN112114134B (en) * | 2019-06-19 | 2024-02-02 | 爱科来株式会社 | Target substance detection method, target substance detection kit, and target substance detection system |
| CN111044729A (en) * | 2019-12-13 | 2020-04-21 | 西安医学院 | Aptamer-based high-sensitivity C-reactive protein nano immunochromatography detection method |
| CN111044729B (en) * | 2019-12-13 | 2023-05-02 | 西安医学院 | Nucleic acid aptamer-based high-sensitivity C-reactive protein nano immunochromatography detection method |
| CN113174447A (en) * | 2021-03-26 | 2021-07-27 | 江苏省农业科学院 | Gold-labeled rapid detection card for detecting CGMMV and preparation method thereof |
| CN114441521A (en) * | 2022-01-27 | 2022-05-06 | 宜宾五粮液股份有限公司 | Preparation method of white spirit ethyl carbamate quick-detection test paper |
| CN116008542A (en) * | 2022-10-21 | 2023-04-25 | 青岛农业大学 | Aptamer test strip, preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103048449B (en) | 2015-06-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103048449B (en) | Chromatographic detection kit based on aptamer, as well as preparation method and detection method thereof | |
| Ching | Lateral flow immunoassay | |
| JP4934147B2 (en) | Test equipment for rapid diagnosis | |
| EP2574926B1 (en) | Chromatographic kit and chromatography method | |
| JP5694922B2 (en) | Simple membrane assay device | |
| US8999730B2 (en) | Immunochromatography method | |
| Zhang et al. | Gold-platinum nanoflowers as a label and as an enzyme mimic for use in highly sensitive lateral flow immunoassays: application to detection of rabbit IgG | |
| CN104820094A (en) | Immune-chromatographic detection test paper and detection method | |
| CN104502586A (en) | Immunochromatography detection method and test paper | |
| EP2042868A1 (en) | Immunochromatographic test device | |
| KR102133752B1 (en) | Lateral flow assay strip | |
| CN107328938A (en) | Propepsin and helicobacter pylori antibody detection method and its kit | |
| CN102507929A (en) | Signal-enhancement type immunochromatographic gold-labeled test strip and preparation method thereof | |
| CN107271669A (en) | Propepsin, helicobacter pylori antibody and G17 detection method and its kit | |
| JP4179419B2 (en) | Test substance detection method, sensitizer and immunochromatography kit | |
| CN107132359A (en) | Pepsinogen Cgene and PGⅡ detection method and its kit | |
| CN104991058A (en) | Preparation method for gold labeled immucochromatographic test strip jointly marked through colloidal gold and latex microsphere | |
| CN106353503A (en) | Novel quality control method applied to immunochromatography test paper strip | |
| CN101957369A (en) | The enhancing mark conjugate that is used for immunoassays | |
| JP5545683B2 (en) | Immunochromatographic test equipment | |
| CN110231489A (en) | A method of improving the test strip biosensor detection sensitivity of carbon nanotube label | |
| JP5782533B2 (en) | Chromatograph method and chromatograph kit | |
| WO2016175336A1 (en) | Immunochromatography device having reduced background noises, and method for reducing background noises in immunochromatographty device | |
| CN104749358A (en) | Semiquantitative immunochromatographic test method and test paper | |
| CN103728452B (en) | Colloidal gold immunoassay kit for detecting aflatoxin B1 and preparation method of kit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |