CN102636376A - Solution for detecting analyte in sample - Google Patents
Solution for detecting analyte in sample Download PDFInfo
- Publication number
- CN102636376A CN102636376A CN2012100738632A CN201210073863A CN102636376A CN 102636376 A CN102636376 A CN 102636376A CN 2012100738632 A CN2012100738632 A CN 2012100738632A CN 201210073863 A CN201210073863 A CN 201210073863A CN 102636376 A CN102636376 A CN 102636376A
- Authority
- CN
- China
- Prior art keywords
- cyclodextrin
- solution
- sample
- beta
- analyte
- 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.)
- Pending
Links
Images
Abstract
The invention provides a solution for detecting whether an analyte exists in a sample or not. The solution comprises cyclodextrin or ramification of cyclodextrin, and a buffering system playing a role in buffering. The solution can be used for improving the sensitivity of narcotics detection, especially the detection aiming at tetrahydrocannabinol (THC), and obviously improving the effect of the sensitivity.
Description
Technical field
The invention belongs to disease diagnosing system, special, the present invention relates to a kind of detection system whether the tracer liquid sample exists the solution of analyte and comprise this solution that is used for.
Background technology
Drugs detect at present by the conventional sense means of most countries as necessity, particularly in registration, drive or look in the malicious process and usually need test personnel.Saliva does not have the detection of wound more and more universal as one to the detected person; And the analyte in the saliva; For example drug abuse is not easy because concentration is little to be detected, and also has some drugs micromolecule in addition, and the utensil that is collected sample easily is adsorbed; Thereby increased the difficulty that detects, caused omission easily.
This just need provide a better improved method that the sample of these collections is handled, and to improve the sensitivity that detects, avoids some defectives in the conventional art simultaneously.
Summary of the invention
In order better to solve above prior art problems, on the one hand, the present invention provides a kind of solution that is used for detecting the sample analyte, comprising: the buffer system of the derivant of cyclodextrin or cyclodextrin, a buffer action.
On the other hand, the present invention provides a kind of being used for to detect the system whether sample exists analyte, comprising: contain the solution system of the derivant of cyclodextrin or cyclodextrin, and applicator, it comprises the parts that absorb sample.
On the other hand; The present invention also provides a kind of method that whether has analyte in the sample that detects; This method comprises: collect liquid sample with gatherer; Let the solution of the derivant that contains cyclodextrin or cyclodextrin contact disease with gatherer and carry out wash-out, detect the liquid that elutes from gatherer, obtain analyte content.The mode that wherein detects can detect with testing element.
Preferably, above-described cyclodextrin is selected from: one or more in methyl flamprop, beta-schardinger dextrin-, alpha-cyclodextrin, methyl-beta-schardinger dextrin-, HP-, hydroxyethyl-, sulphur butyl-beta-schardinger dextrin-, carboxymethyl-beta-cyclodextrin, maltodextrin, piroxicam-beta-schardinger dextrin-, the sulfobutyl ether-beta-cyclodextrin.Preferred, described cyclodextrin is one or more in methyl flamprop, beta-schardinger dextrin-, alpha-cyclodextrin, the methyl-beta-schardinger dextrin-.Preferred, the mass percent concentration of described cyclodextrin is at least 0.02-5%.
As preferred scheme, described buffer system is selected from one or more in phosphate, the acetic acid.
Preferably, described solution also comprises anti-THC or Δ
9The antibody of-THC.Preferably, antibody and biotin coupling join.
Preferably, described solution also wraps BSA, and wherein BSA and biotin coupling join.
Preferably, the parts of absorption sample comprise filter paper, nonwoven fabrics or sponge.Preferred, the parts that absorb sample are filter paper.
On the other hand, the present invention also provides a kind of testing element that detects analyte in the sample, and this testing element comprises: surveyed area, and the sample that is positioned at the surveyed area upper reaches applies the zone; Surveyed area comprises a kind of specific bond molecule that is fixed, and wherein, also comprises the cyclodextrin that exists with the form of doing or the derivant of cyclodextrin at the upper reaches of surveyed area.Wherein detecting element can be a test strip.Preferably, in testing element, let liquid sample earlier with testing element on cyclodextrin or the derivant of cyclodextrin contact suitable time formation mixing material, and then let mixing material flow in the surveyed area.The suitable time can be 10 seconds-30 minutes, can be 1,2,3,4,5,6,10,15 or 20 minute.
Preferably, testing element comprises marked region, and marked region applies between zone and the surveyed area at sample.Preferred, marked region comprises antibody and the colloid gold particle of anti-THC; Molecule fixing on the surveyed area comprises THC antigen or haptens.
Preferably, the derivant of cyclodextrin or cyclodextrin is positioned at sample and applies on the zone.
On the other hand, the present invention provides a kind of device of collecting liquid sample, wherein on gathering-device, comprises the cyclodextrin that exists with the form of doing or the derivant of cyclodextrin.Preferably, in this device, let liquid sample earlier with gathering-device on the derivant of cyclodextrin or cyclodextrin contact and carry out wash-out; Test the content of analyte in the eluent then.
On the other hand, the present invention provides the method in a kind of tracer liquid sample, and this method comprises: let liquid sample contact with the derivant of cyclodextrin or cyclodextrin earlier and form mixing material, and then make this mixing material to be detected, thereby obtain testing result.The derivant of wherein said cyclodextrin or cyclodextrin exists in the buffering liquid or with the form of doing and exists on the carrier.Described carrier comprises on the water absorptivity porous carriers such as filter paper, spun glass element, nonwoven fabrics.
Beneficial effect
Through the present invention, can improve the sensitivity that drugs detect, avoid omission simultaneously.
Description of drawings
Figure 1A is the perspective view of test strip in the embodiment of the present invention;
Figure 1B is the plan structure synoptic diagram of test strip in the embodiment of the present invention;
Description of reference numerals: test strip 101; Marker slip 121, sample receiving sheet 111, detection lug 131, suction sheet 141; Support chip 151; Detection line 131; Testing result control line 133.
Embodiment
Further explanation done in the structure or these the employed technical terms that reach in the face of the present invention down.
Cyclodextrin
Cyclodextrin be six above D-glucopyranose units obtaining from starch with alpha-1, the compound sugar irreducibility series of compounds that the 4-glycosidic bond links with the hydrophobic pyramid type structure of ring-type (cyclodextrin model of upper right rotation).In this cyclodextrin series; Their name is based on the quantity of glucosyl group in the ring, and the cyclodextrine so that 6 D-glucopyranose units are formed is called alpha-cyclodextrin (alpha-CD); By that analogy; 7 be called beta-schardinger dextrin-(beta-CD), 8 be called gamma-cyclodextrin, (gamma-CD).More than these three kinds of cyclodextrin be the most frequently used, we claim that also above these three kinds of cyclodextrin are the foundation ring dextrin.Process is carried out after chemical modification and the modification these three kinds of foundation ring dextrin, and we are referred to as the derivant of foundation ring dextrin.The derivant that also is called cyclodextrin.Villiers has found cyclodextrin at first in 1891, he isolates a kind of crystalline material (being called as cyclodextrin afterwards) with bacillus through nutrient culture media from potato starch.Now; The bacterium of commercial production cyclodextrin is to use a kind of natural enzyme; Cyclodextrin glucose transferase (CGTase) is cut off from two ends behind the spiral starch to couple together it and is formed a hydrophobic pyramid type ring-type; Because it is different to cut the starch length of coming, and has so just produced three kinds of different cyclodextrines.
Cyclodextrin is a series of member cyclic oligosaccharides that starch forms after the effect of cyclodextrin glucose base translocase; Commonly used is respectively by 6,7,8 a-cyclodextrin that glucose molecule constitutes; Beta-schardinger dextrin-and γ one cyclodextrin, wherein the purposes with β one cyclodextrin is at present the widest again.The derivant that can comprise above cyclodextrin or cyclodextrin in the solution among the present invention or the decorum.In some preferred modes, β one cyclodextrin or the composition that is used as among the present invention with the derivant that β one cyclodextrin is the basis use, and can reduce preferably to absorb the absorption of the parts of sample to analyte on the sampler, for example to the absorption of THC.For example, one or more in methyl-beta-schardinger dextrin-, HP-, hydroxyethyl-, sulphur butyl-beta-schardinger dextrin-, carboxymethyl-beta-cyclodextrin, piroxicam-beta-schardinger dextrin-, the sulfobutyl ether-beta-cyclodextrin.Preferred, described cyclodextrin is methyl flamprop, beta-schardinger dextrin-, alpha-cyclodextrin, methyl-beta-schardinger dextrin-.These are limited enumerating for example, scope of the present invention are not constituted any restriction.
As the derivant of cyclodextrin that exists with solution or cyclodextrin, solvent can be any its dissolving, suspension reagent wherein of can letting such as water.In some preferred modes, comprise also in the solution that some play the reagent of buffer action, for example phosphoric acid, hydrochloric acid, acetic acid, Tris-hydrochloric acid or the like.The such reagent that exists with the solution form can be better with sample in analyte contact or let analyte elute from the sampling carrier.The sampling carrier comprises in ST, the ST that the parts that absorb sample can make any carrier that can retain sample, and carrier comprises any porous carrier or non-suction carrier that can the absorption liquid sample body.The suction porous carrier comprises for example sponge, cotton, filter paper, nonwoven fabrics; Perhaps non-suction carrier comprises plastics, glass etc.
It also is the another one factor that the concentration of cyclodextrin is selected, and can select different concentration according to different kinds, and for example accounting for the eluent percentage by weight is 0.05-5%, 0.2-3%; 0.2-2.5%; 0.2-2.0%; For example, when the cyclodextrin of selecting was methyl-beta-schardinger dextrin-, the concentration of methyl-beta-schardinger dextrin-was 0.2-5% in the eluent, and preferably 0.2-3% for example is 0.1%, 0.5%, 0.25%, 0.6%, 0.8%; 2.5%; 3.0% etc.Above embodiment particularly suitable wash-out gatherer is to the absorption of THC.If when being directed against other analyte certainly; Can confirm to use the kind of cyclodextrin and the concentration that variety classes needs through some simple tests; This is that one of ordinary skill in the art is expected through reading the present invention easily, and identifiable through the limited number of time test.
When the derivant of cyclodextrin or cyclodextrin (is called again: when eluent) existing with solution; A kind of method is provided; Let sample contact earlier or let the gatherer that has sample contact and carry out wash-out with elute soln with elute soln; Then to testing, thereby obtain test result from the solution of gatherer wash-out.Test comprises with testing element to be tested, and also comprises with the general instrument of liquid chromatography, gas chromatography or matter and testing.The mode of wash-out can be to let the solution of the derivant that contains cyclodextrin or cyclodextrin directly contact a period of time with gatherer, also can let gatherer extruding repeatedly in elute soln, and rinsing perhaps lets suitable time of elute soln flushing gatherer or the like.
On the other hand; These above cyclodextrin not only can exist with the form of solution, can also directly handle on pick-up unit and with the form of doing to exist, and for example exist on some testing elements and with the form of doing; Testing element comprises test strip, for example on the nitrocellulose filter.For example, be configured to solution to cyclodextrin, the sample at testing element applies the dextrin solution of handling certain volume on the zone then, then oven dry.When testing, let liquid sample and test unit see that the zone of going up the solution of handling the derivant that cyclodextrin or cyclodextrin are arranged contact earlier and form mixing material, and then let mixing material flow on the first surveyed area of seeing of test.Test through us finds, such mode can reduce also that sample applies the zone because the difference of material and to the absorption of some small-molecule substances, for example to the absorption of THC molecule.
Therefore, the present invention provides a kind of testing element, comprises that the sample that is positioned at the test zone upper reaches applies the zone; Surveyed area, it comprises the specific bond molecule that is fixed on surveyed area; Wherein, comprise the cyclodextrin molecular or derivatives thereof that exists with the state of doing at the upper reaches of surveyed area.On the other hand, the present invention also provides a kind of device of collecting liquid sample, and wherein, this device comprises the cyclodextrin molecular or derivatives thereof that exists with the state of doing.This gathering-device comprises with spongy top, and the parts of the absorption liquid sample body that materials such as filter paper are formed comprise the cyclodextrin molecular or derivatives thereof on the parts of these absorption samples.The variety classes of the cyclodextrin of above-mentioned elaboration and concentration can be included in these some concrete embodiments.
Sample
The sample of any kind can both make an experiment with device of the present invention, comprises body fluid (for example, urine and other body fluid, and clinical sample).Liquid sample possibly be derived from solid or semisolid sample, comprises excrement, biological tissue and food sample.These solids can be transformed into liquid sample through any suitable method with semisolid sample, for example in a kind of suitable liquid, mix, stamp broken; Macerate, hatch, dissolving or enzymolysis solid sample are (for example; Water, phosphate buffer or other damping fluid)." biological specimen " comprises the sample that is derived from animal alive, plant and food; Also comprise urine, saliva, blood and blood constituent, cerebrospinal fluid, vaginal swab; The culture of seminal fluid, ight soil, sweat, secretion, tissue, organ, tumour, tissue and organ; Condition medium cell culture and there is no matter be the people's or animal.Food sample comprises finished COF and last product, meat, cheese, wine, milk and potable water.Plant sample comprises the sample of the condition medium that is derived from any plant, plant tissue, plant cell cultures and there." environmental samples " is those samples that are derived from environment (for example, the sample of lake water sample or other water body, sewage sample, soil sample, underground water sample, seawater sample, the samples of runoff water).Sewage also can be included in the environmental samples with relevant refuse.
Analyte
Can analyze any analyte with the present invention.The example of the analyte of the enough stable detection of the present invention of ability comprises (but not only comprising) human chorionic gonadotrophin (hCG); Lutropin (LH); Ovarian stimulation plain (FSH), hepatitis C virus (HCV), hepatitis B (HBV); Hepatitis B surface antigen, the medicine of AIDS virus and any abuse.Analyte can or liquefy at any liquid and detect in the sample, urine for example, saliva, saliva, blood, blood plasma, perhaps serum.The example of other analyte also has the acid of flesh ammonia acid anhydride, cholerythrin, nitrite, protein (nonspecific), blood; Leucocyte, blood sugar, heavy metal and toxin, bacterium composition (for example, special protein and the sugar of the bacterium of particular type; Colon bacillus 0157: H7 for example, staphylococcus aureus, salmonella, C.perfringens; Campylobacter, listeria monocytogenes, enteritis vibrios, perhaps cured shape bacillus).Any other analyte of suitable lateral flow assay form can detect with this device.Analyte can also be the material that material maybe can point out infection period that is infectious.Analyte can be medicine (like a drug abuse), hormone, albumen, nucleic acid molecules, pathogen." drug abuse " (DOA) is meant that the non-medical destination uses medicine (playing the paralysis nerve usually).Abuse these medicines and can cause body & mind to suffer damage, produce dependence, habit-forming and/or dead.The example of drug abuse comprises cocaine; Amphetamine (for example, black beauty, white amphetamine tablet, dextroamphetamine, dexie, Beans); Crystal methamphetamine (crank, meth, crystal, speed); Barbiturate is (like
Roche Pharmaceuticals; Nutley, New Jersey); Sedative (paramedicines of promptly sleeping); Lysergic acid diethylamide (LSD); Suppressant (downers, goofballs, barbs, blue devils, yellow jackets, methaqualone); The anti-antidepressant of tricyclic antidepressants (TCA, i.e. imipramine, amitriptyline and doxepin); Hog (PCP); Tetrahydrocannabinol (THC, pot, dope, hash, weed, etc.), Δ
9-THC; Opiate (being morphine, opium, codeine, heroin, the hydroxyl dihydrocodeinone) and their metabolic product in animal body.Use this test strip also can be used to belong to medical usage but the detection of overdose easily, like tricyclic antidepressant (imipramine or analog) and Paracetamol.
Testing element
Testing element can be selected the test strip of cross flow for use, and it can detect multiple analyte.Certainly, other suitable testing elements also can be used in the present invention.Various testing elements can be combined in together and apply among the present invention.A kind of form is to detect test paper.The detection test paper that is used for the analyte (like drugs or show the metabolin of health) of analyzing samples can be a various forms, like immunoassays or chemico-analytic form.Detect the analytical model that test paper can adopt non-competing method or competition law.Detect test paper generally comprise one have sample application zone absorbent material, reagent area and test section.Add sample to sample application zone, flow to reagent area through capillarity.At reagent area, if there is analyte, sample combines with reagent.Sample continues to flow to detection zone then.Other reagent are fixed on detection zone like the molecule that combines with the analyte specificity.Analyte in these reagent and the sample (if existence) reacts and analyte is combined in this district, perhaps combines with the some reagent of reagent area.The label existence that is used to show detection signal and reagent area or the mark zone that separates.
Typical non-competing method analytical model is that signal will produce if contain analyte in the sample, if do not comprise analyte, does not just produce signal.In competition law, if analyte is not present in the sample, signal produces, if there is analyte, does not then produce signal.
Testing element also can be to detect test paper, can select the material of suction or non-suction for use.Detect test paper and can comprise that multiple material is used for the liquid sample transmission.Wherein a kind of material that detects test paper can cover on the another kind of material, covers on the nitrocellulose filter like filter paper.A district detecting test paper can select one or more materials for use, and another district selects other one or more different materials for use.The detection test paper can be attached on certain holder or hard surface is used to improve the intensity of being affectedly bashful the detection test paper.
Analyte is detected through signal generating system; Like utilization one or more enzymes with this analysis thing generation specific reaction; Utilize as aforementioned specific combinating substance is fixed on the method that detects on the test paper, the composition of one or more signal generating systems is fixed on the analyte detection zone of detection test paper.The material that produces signal can be in sample application zone, reagent area, or detection zone, or on the whole detection test paper, this material can be full of on one or more materials that detect test paper.The solution that will contain the signal thing is added to the surface of test paper or one or more materials of test paper is immersed in the solution that contains the signal thing.Make and add the test paper drying that contains signal thing solution.
Each district of detecting test paper can arrange by following mode: sample application zone, reagent area, detection zone.Or be necessary to comprise the control zone, and the zone whether mingled of definite sample, the liquid sample uptake zone.The control zone is positioned at after the detection zone.All districts only can be arranged on the test paper with a kind of material.Also but material different is not adopted in the same district.Each district can directly contact with liquid sample, or different districts arranges according to the direction that liquid sample flows, and the end in Jiang Ge district links to each other with the front end in another district and overlaps.Used material can be water absorptivity material such as a filter paper preferably, glass or nitrocellulose filter etc.Detect test paper and also can adopt other forms.
Be used for disclosed testing element and can be any known testing element of prior art, such testing element includes, but are not limited to: the immunoassays of being known in the prior art, and the test of chemical assay and enzyme, such as, but be not limited to; The monospecific antibody immunoassays, multiple antibody mediated immunity is measured, and complex immunity is measured, the immunoassays of contrast; Immunoassays of non-contrast or the like comprise and utilize HRPO, alkaline phosphatase, luciferase; The antibody pairing, antibody fragment, FLA, modified antibody; Labelled antibody, the antibody of colloid gold particle, or with antibody of painted gel particle mark or the like, they all are to know in the prior art.The instance that can be included into some testing elements of this device can find in the United States Patent (USP) below: US 4857453; US 5073484; US 5119831; US 5185127; US 5275785; US 5416000; US5424193; US 5504013; US 5602040; US 5622871; US 5654162; US 5656503; US 5686315; US 5766961; US 5770460; US 5916815; US 5976895; US 6248598; US 6140136; US 6187269; US 6187598; US 6228660; US 6235241; US 6306642; US 6352862; US 6372515; US 6379620; US 6403383; US6656744; US6979576; US6372513; US 6372513.The instance that further, can be included into some testing elements of this device can find in the U.S. Patent application below: sequence number is 09/579672; 09/579673; 09/653032; 60/233739; 09/915494,10/211199 and 09/860408.
Detect
Detect the expression chemical examination or test a kind of material or whether material exists; Such as; But be not limited to this, the metabolin of chemical substance, organic compound, mineral compound, metabolism product, medicine or drug metabolite, organic organization or organic organization, nucleic acid, protein or polymkeric substance.In addition, detect the quantity of expression test substances or material.Furtherly, immune detection, chemical detection, enzyme detection etc. are also represented in chemical examination.
Embodiment
For how clearer elaboration the present invention realizes, now describe with limited experiment, limited giving an example only done in these explanations under the marrow of claim of the present invention, do not constitute the restriction to claim of the present invention.Do not specialize, the used number percent of the present invention is mass percent.
Implement 1: have methyl-beta-schardinger dextrin-(RMCD) in the damping fluid and do not exist methyl-beta-schardinger dextrin-(RMCD) to detecting
The comparison of hemp prototype in the saliva.
Material:
Saliva collector-40 sponge stick, 40 detection test paper are our company and produce voluntarily, choose the product of same lot number; Basis buffering liquid: phosphate buffer solution (PH=8.3); Colorimetric card.
Step:
1. dispose the 100ng/ml Δ
9The positive hemp primary standard of-THC article
2. each on 20 saliva collectors adds 3 milliliters of 100ng/ml Δs
9Positive hemp prototype (or cannabinol) standard solution of-THC;
3. be configured to A in the table 1 with basic damping fluid, two kinds of different buffering liquids of B are contained in these damping fluids respectively in 10 vials then; 3 milliliters of buffer solution of each vial splendid attire; Be inserted into saliva collector in these vials then, let the buffering liquid in the vial flood the cavernous body that absorbs liquid as far as possible, push about 10 times then back and forth; Abandon saliva collector, (A handles: 10 parts to obtain totally 20 parts of eluting liquids respectively; B handles: 10 parts);
4. use basic damping fluid and 3 milliliters of 100ng/ml Δs of 3 milliliters in addition
9The mixing of the positive hemp primary standard of-THC article obtains C and handles: 10 parts; In addition with the basic damping fluid that contains 1%RMCD and 3 milliliters of 100ng/ml Δs of 3 milliliters
9The mixing of the positive hemp primary standard of-THC article obtains D and handles: 10 parts;
Collect above four in liquid drip 100 microlitres separately and apply the zone to the sample of detection test paper, each handles 10 test strip; Whether the detection zone that detects by an unaided eye after 10 minutes has lines to occur, and comes relatively to detect the shade degree of lines with the colour atla (table 2) of standard.And compare interpretation testing result ("+" expression is positive, and "-" expression is negative) with the colour atla of standard.
6. the preparation of test strip is prepared according to following method:
6.1. the processing of nitrocellulose filter Detection lug 131 is nitrocellulose filter (NC), and two lines are arranged on nitrocellulose filter, and one is detection line 207, is positioned at the testing result control line 206 in detection line downstream.On detection line 132, be fixedly connected with the Streptavidin (streptavidin-IgG) of IgG; Fixing goat anti-rabbit igg on testing result control line 133.Fixed method can use automatic spray film handling machine to handle automatically, and wherein the concentration at the processing and detecting lines is 0.3 mg/ml, and dilution buffer liquid is phosphate buffer (PBS); Fixing goat anti-rabbit igg antibody on the testing result control line; Concentration is 0.3 mg/ml, and dilution buffer liquid is phosphate buffer (PBS).Be placed on the nitrocellulose filter of handling well to dry in 37 ℃ of baking ovens and get final product.
6.2 the processing of marker slip 121Marker slip is the polyester film, and the THC haptens that have coupled protein molecule BSA good by the colloid gold particle mark is processed on the polyester film.The OD value of Treatment Solution is 75, and dilute solution is 1 times of PBS buffer solution, wherein also contains 1% BSA.On marker slip, also handle the rabbit igg antibody that colloid gold label is arranged.Be placed on the marker slip of handling well to dry in 37 ℃ of baking ovens and get final product.
6.3 the processing of sample blank film 111The sample blank film is the plain film of spun glass, and the solution of on this film, handling is: Borax (0.07M/L); Tween20 (1%); Cholic Acid (1%); Tris (0.1M).Be placed on the sample blank film of handling well to dry in 37 ℃ of baking ovens and get final product.
6.4. the assembling of reagent stripAccording to the assembling of the appearance shown in Figure 1B, let each parts of handling well the sample blank film be positioned at the upper reaches of marker slip, marker slip is placed the filter paper of suction in the downstream of detection lug between detection lug and sample blank film.These parts all are placed on the non-absorptive support chip.
Test result such as following table 1
Table 1 contains cyclodextrin and does not contain the influence of cyclodextrin to analyte THC adsorption effect
Can find out obviously that by last table after the sponge stick sampling, the sponge of sponge stick obviously shows the absorption to the hemp prototype, makes testing result negative, does not meet actual conditions.And use buffer elution liquid of the present invention can wash away the THC that is attracted on the sponge, elute effect that can clearer reflection damping fluid.It is thus clear that, contain the most of down hemp component that adsorbs of elution buffer wash-out of 1%RMCD, thereby can obviously improve the detection sensitivity of hemp, can not influence the net result of whole detection in addition yet.
In addition; The present invention has carried out similar experiment to the derivant of some cyclodextrin simultaneously; For example; Methyl flamprop, beta-schardinger dextrin-, alpha-cyclodextrin, methyl-beta-schardinger dextrin-, HP-, hydroxyethyl-, sulphur butyl-beta-schardinger dextrin-, carboxymethyl-beta-cyclodextrin, piroxicam-beta-schardinger dextrin-, sulfobutyl ether-beta-cyclodextrin have obtained essentially identical conclusion, and concrete experimental data slightly.
Table 2: colour atla standard and testing result (based on competition law)
2 different RMCD concentration ratios are implemented in
.
Material:
Saliva collector: 40 sponge sticks, 40 detection test paper are our company and produce voluntarily, choose the product of same lot number.
Basis buffering liquid: phosphate buffer solution (PH=8.3);
Colorimetric card
Step:
1. dispose the 100ng/ml Δ
9The positive hemp primary standard of-THC article;
2. each on 100 saliva collectors adds 3 milliliters of 100ng/ml Δs
9The positive hemp primary standard article solution of-THC;
3. be configured to table 10 kind of different buffering liquid with basic damping fluid, wherein the concentration of RMCD is respectively with containing 0,0.1%, 0.2%, 0.5%, 1.0%, 1.5%, 2.0%, 2.5%, 5%, 10%; Be contained in every kind of damping fluid respectively in 10 vials then; 3 milliliters of buffer solution of each vial splendid attire; Be inserted into saliva collector in these vials then, let the buffering liquid in the vial flood the cavernous body that is absorbed with liquid sample as far as possible, push back and forth then about 10 times; Abandon saliva collector, obtain totally 100 parts of eluting liquids respectively;
4. drip 100 microlitres to above 10 kinds of liquid of collecting separately and apply the zone to the sample that detects test paper (the same with the detection test paper in the examples of implementation 1), every kind of eluent is handled 10 test strip; Whether the detection zone that detects by an unaided eye after 10 minutes has lines to occur, and comes relatively to detect the shade degree of lines with the colour atla (table 2) of standard.And compare interpretation testing result ("+" expression is positive, and "-" expression is negative) with the colour atla of standard.
5. control experiment: except there is not any Δ in standard variety
9Outside-the THC, other method is the same with above step 1-4.
Table 3: different RMCD concentration compare the elute effect of THC in the saliva
It is thus clear that for THC positive criteria article, the RMCD of 0.2% above concentration all has comparatively significantly elute effect; And for the negative standard items (PBS) of THC, be higher than 5% RMCD concentration will cause detection signal to weaken significantly, unfavorable interpretation as a result, higher concentration possibly cause false positive results.
The comparison of when
enforcement 3 different sampling materials are collected sample as absorption analyte being adsorbed and the comparison of elute effect.
As the sampling instrument that absorbs saliva sample, drip the 100ng/ml Δ of certain volume with material in the following tabulation 4 on the sample collection parts that unlike material is formed in table 4 respectively
9The positive hemp primary standard article of-THC also delay with isopyknic wash-out that (one is to comprise the phosphoric acid eluent that contains 1%RMCD; Another is basic phosphate buffer: PBS; PPH=8.3) carry out wash-out towards liquid, the method for wash-out is referring to the method among the embodiment 2, and each handles repetition 10 times.
Drip 100 microlitres to above 5 kinds of liquid of collecting separately and apply the zone to the sample that detects test paper (the same with the detection test paper in the examples of implementation 1), every kind of eluent is handled 10 test strip; Whether the detection zone that detects by an unaided eye after 10 minutes has lines to occur.
Table 4: the comparison of when different sampling materials are collected sample as absorption analyte being adsorbed
It is thus clear that filter paper is comparatively desirable as the sampling material, no matter under PBS still was the eluent situation, the absorption situation was ideal for it.
Implement 4: the comparison of different effluent volume differences under the same concentrations
When with filter paper as sampling instrument, on filter paper, drip the 100ng/ml Δ
9The positive hemp primary standard of-THC article are also used for 1 milliliter and are contained the different effluent volume of 1%RMCD, are respectively the 1x of filter paper sampling rod, 2x, and 3x, 4x, the 5x volume, the method for wash-out is the same with the elution process of a kind of examples of implementation.
Drip 100 microlitres to above 5 kinds of liquid of collecting separately and apply the zone to the sample that detects test paper (the same with the detection test paper in the examples of implementation 1), every kind of eluent is handled 10 test strip; Whether the detection zone that detects by an unaided eye after 10 minutes has lines to occur.
The result is as follows:
Table 5: the comparison of different effluent volume differences under the same concentrations
Visible among the result, the eluent of 1-3 times of volume all can obtain positive findings, and if consider that the difficulty or ease of wash-out operation and subsequent sample confirm required volume, the eluent of 2-3 times of volume should be proper.
Implement 5: the comparison of different types of cyclodextrin elute effect
Δ-THC standard items of 100ng/ml of 3 milliliters progressively increase on spongy top; Carry out wash-out according to the elution process among the embodiment 1 then; The volume of every kind of eluent is 3 milliliters; Comprise in the eluent that final concentration is 3 kinds of different cyclodextrin of 1%: methyl-beta-schardinger dextrin-(RMCD), HP-(HPCD), beta-schardinger dextrin-(β-CD).After pushing 10 times repeatedly then, abandon spongy top, drip 100 microlitres to above 3 kinds of liquid of collecting separately and apply the zone to the sample that detects test paper (the same with the detection test paper in the examples of implementation 1), every kind of eluent is handled 10 test strip; Whether the detection zone that detects by an unaided eye after 10 minutes has lines to occur, and is contrast with basic buffering liquid.
Table 6: the comparison of different types of cyclodextrin elute effect
| Handle | Contrast | RMCD | HPCD | ?β-CD |
| Signal intensity | G8 | G2 | G4 | ?G7 |
It is thus clear that HPCD also has anti-absorption property, but than a little less than the RMCD, and the anti-suction-operated of β-CD is not so good as RMCD, HPCD is obvious.
The antibody of implementing to contain in 5 damping fluids anti-THC of Biotin-is coupled thing and is coupled with the antibody that does not contain the anti-THC of Biotin-
THC must influence thing in the sample to detecting.
Utilize the filter paper sampling rod to collect the 100ng/ml Δ for preparing in advance
9The positive hemp primary standard of-THC article, each absorbs 1 milliliter of standard items, and sampling rod usefulness contains the elution buffer that Biotin-THC Ab is coupled thing and carries out wash-out, and eluent detects with agent plate; Carry out control experiment simultaneously, sampling rod does not carry out wash-out with not containing the elution buffer that Biotin-THC Ab is coupled thing, and the eluent of collection adds and contains the Biotin-THC Ab that Biotin-THC Ab is coupled the elution buffer moderate of thing and be coupled thing.
Drip 100 microlitres to above 2 kinds of liquid of collecting separately and apply the zone to the sample that detects test paper (the same with the detection test paper in the examples of implementation 1), every kind of eluent is handled 10 test strip; Whether the detection zone that detects by an unaided eye after 10 minutes has lines.
Table 6: elution buffer contains Biotin-THC Ab and is coupled thing and does not contain Biotin-THC Ab and be coupled thing to detecting the influence of THC in the sample
It is thus clear that, make target hemp antigen and Biotin-THC Ab be coupled thing reaction in the elution process and generate antigen antibody complex, can effectively be reduced in wash-out and the testing process since hemp with contact material and adsorb the situation that causes the detection sensitivity reduction each other.
Implement 6: when containing Biotin-BSA in the damping fluid and being coupled thing, Biotin or Biotin derivant detection signal is kept
Between experiment.
Utilize the filter paper sampling rod to collect the 100ng/ml Δ for preparing in advance
9The positive hemp primary standard of-THC article, each sampling rod is collected 1 milliliter of standard solution; Sampling rod carries out wash-out with containing the elution buffer that Biotin-BSA is coupled thing, Biotin or Biotin long-chain derivative respectively, and the volume of eluent is 1 milliliter.Detect with test strip, investigate the signal stabilization in testing result and a period of time.
The same in present embodiment among test strip and the embodiment 1.
Table 7: experimental result
It is thus clear that biotin is not added in contrast, signal just begins to strengthen gradually in back 30 minutes of detection; After 120 minutes, show reinforcing yin essence property; But shortened the read time of testing result, be unfavorable for greatly using in the user, the experimental result that reads in different time in addition is also inaccurate.And be coupled thing, Biotin or Biotin long-chain derivative through adding Biotin-BSA, can keep that signal intensity does not change in 2 hours.
Claims (18)
1. one kind is used for detecting the solution whether sample exists analyte, comprising: the buffer system of the derivant of cyclodextrin or cyclodextrin, a buffer action.
2. solution according to claim 1, wherein, described cyclodextrin is selected from: a-cyclodextrin, one or more in beta-schardinger dextrin-and the gamma-cyclodextrin.
3. solution according to claim 1, wherein, described cyclodextrin is a beta-schardinger dextrin-.
4. solution according to claim 3; Wherein, described beta-schardinger dextrin-is one or more in methyl-beta-schardinger dextrin-, HP-, hydroxyethyl-, sulphur butyl-beta-schardinger dextrin-, carboxymethyl-beta-cyclodextrin, piroxicam-beta-schardinger dextrin-, the sulfobutyl ether-beta-cyclodextrin.
5. solution according to claim 1, wherein, described buffer system is selected from phosphate, acetic acid, one or more in the Tris-hydrochloric acid.
6. solution according to claim 1, wherein, described solution also comprises anti-THC or Δ
9The antibody of-THC, wherein antibody and biotin coupling join.
7. solution according to claim 1, wherein, described solution also comprises BSA, wherein BSA and biotin coupling join.
8. according to the described solution of one of claim 1-7, wherein, the mass percent concentration of described cyclodextrin is 0.2-5%.
9. one kind is used for detecting the system whether sample exists analyte, comprising: include cyclodextrin or cyclodextrin derivant solution system and comprise applicator, it comprises the parts that absorb sample.
10. system according to claim 9, wherein, the parts that absorb sample comprise filter paper, nonwoven fabrics or sponge.
11. system according to claim 10, wherein, the parts that absorb sample are filter paper.
12. system according to claim 9, wherein, described solution also comprises anti-THC or Δ
9The antibody of-THC, wherein antibody and biotin coupling join.
13. system according to claim 9, wherein, described solution also wraps BSA, and wherein BSA and biotin coupling join.
14. system according to claim 9, wherein, the mass percent concentration of described cyclodextrin is at least 0.2-5%.
15. system according to claim 9, wherein, described cyclodextrin is one or more of beta-schardinger dextrin-, alpha-cyclodextrin and gamma-cyclodextrin.
16. one kind is used for detecting the detection test paper whether sample exists analyte, wherein this test paper comprises: surveyed area, and be positioned at the surveyed area upper reaches and the cyclodextrin that exists with the form of doing or the derivant of cyclodextrin.
17. one kind is used for detecting the method whether sample exists analyte, comprises: collect liquid sample with gatherer; Let this gatherer of eluant solution of the derivant that contains cyclodextrin or cyclodextrin; Analyte in the test eluent.
18. method according to claim 17, wherein, with the analyte in the testing element test eluent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012100738632A CN102636376A (en) | 2012-03-20 | 2012-03-20 | Solution for detecting analyte in sample |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012100738632A CN102636376A (en) | 2012-03-20 | 2012-03-20 | Solution for detecting analyte in sample |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102636376A true CN102636376A (en) | 2012-08-15 |
Family
ID=46620855
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012100738632A Pending CN102636376A (en) | 2012-03-20 | 2012-03-20 | Solution for detecting analyte in sample |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102636376A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106093421A (en) * | 2016-05-31 | 2016-11-09 | 常州博闻迪医药科技有限公司 | A kind of saliva Procalcitonin. fluorescence immunoassay detection method |
| CN109187921A (en) * | 2018-08-31 | 2019-01-11 | 长江大学 | A kind of semi-automatic disposal suitable for ammonia volatilization experiment |
| US11485535B2 (en) | 2018-12-19 | 2022-11-01 | The Procter & Gamble Company | Article with visual effect |
| CN117783507A (en) * | 2024-02-27 | 2024-03-29 | 广州科方生物技术股份有限公司 | Component liquid commonly used in (1-3) -beta-D glucan and galactomannan detection, and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20020001539A1 (en) * | 2000-03-31 | 2002-01-03 | Dicesare Joseph L. | Apparatus and methods for chemiluminescent assays |
| CN101109750A (en) * | 2007-07-27 | 2008-01-23 | 杭州浙大生科生物技术有限公司 | Reagent kit for detecting food allergen specificity IgG ELISA and preparing method thereof |
| CN102338765A (en) * | 2010-12-23 | 2012-02-01 | 深圳大学 | Blood sugar testing paper |
-
2012
- 2012-03-20 CN CN2012100738632A patent/CN102636376A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20020001539A1 (en) * | 2000-03-31 | 2002-01-03 | Dicesare Joseph L. | Apparatus and methods for chemiluminescent assays |
| CN101109750A (en) * | 2007-07-27 | 2008-01-23 | 杭州浙大生科生物技术有限公司 | Reagent kit for detecting food allergen specificity IgG ELISA and preparing method thereof |
| CN102338765A (en) * | 2010-12-23 | 2012-02-01 | 深圳大学 | Blood sugar testing paper |
Non-Patent Citations (2)
| Title |
|---|
| 赵霞 等: "毛细管电泳在毒品检验中的应用", 《刑事技术》, 31 December 2004 (2004-12-31) * |
| 陈龙然 等: "一株产环糊精葡萄糖基转移酶的地衣芽孢杆菌的选育、产酶条件及酶学特性", 《微生物学报》, vol. 45, no. 1, 28 February 2005 (2005-02-28) * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106093421A (en) * | 2016-05-31 | 2016-11-09 | 常州博闻迪医药科技有限公司 | A kind of saliva Procalcitonin. fluorescence immunoassay detection method |
| CN109187921A (en) * | 2018-08-31 | 2019-01-11 | 长江大学 | A kind of semi-automatic disposal suitable for ammonia volatilization experiment |
| US11485535B2 (en) | 2018-12-19 | 2022-11-01 | The Procter & Gamble Company | Article with visual effect |
| CN117783507A (en) * | 2024-02-27 | 2024-03-29 | 广州科方生物技术股份有限公司 | Component liquid commonly used in (1-3) -beta-D glucan and galactomannan detection, and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101876657B (en) | Rapid sample detection and storage devices and methods of use | |
| CN101021526B (en) | Apparatus and method for visual display of detecting result | |
| CN105424931B (en) | Helicobacter Pylori urease antibody IgM, IgG associating device for fast detecting and preparation method thereof | |
| AU2007280929B2 (en) | Analysis device for biological sample | |
| CN103575893A (en) | Method for rapidly detecting shellfish toxin | |
| CN205861689U (en) | A kind of detection device | |
| CN102636376A (en) | Solution for detecting analyte in sample | |
| CN104459125A (en) | Method for rapidly detecting gram negative and positive bacteria | |
| CN203083995U (en) | Sample detection device | |
| Zeng et al. | A simple and rapid immunochromatography test based on readily available filter paper modified with chitosan to screen for 13 sulfonamides in milk | |
| CN105606808A (en) | Group A/B streptococcus colloidal gold immunochromatograohic assay detection device and detection method thereof | |
| Wang et al. | Development of an immunochromatographic test to detect white spot syndrome virus of shrimp | |
| CN108918849A (en) | The quickly method of the aflatoxin in detection medicinal material and test card used | |
| JPH02504188A (en) | Capillary device for multiple analysis immunoassays | |
| CN102087285B (en) | Immunochromatographic test strip for rapidly detecting acute pancreatitis and preparation method thereof | |
| CN102124341A (en) | Rapid detection of post-vaccination antibody response | |
| CN105842464B (en) | Joint based on up-converting phosphor technology quantitatively detects uNGAL and uCr device and preparation method thereof | |
| CN200968955Y (en) | Biological liquid sample analytical equipment | |
| CN101592660A (en) | Brucellosis indirect enzyme-linked immunosorbent assay milk humoral antibody detection kit | |
| CN109342740A (en) | A kind of fluorescence immune chromatography kit and preparation method thereof detecting people's pylori spiral bacilli antibody | |
| CN108918850A (en) | The quickly method of the aflatoxin in detection medicinal material | |
| US11808672B2 (en) | Detection device | |
| CN102004146B (en) | Mixed maker, marking method and application thereof | |
| CN102539755A (en) | Test strip for detecting influenza A virus antigen in secretion and preparation method thereof | |
| CN1453588A (en) | Reagent for colloidal gold chromatographic analysis of SARS coronavirus antigen |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20120815 |