CN1399132A - Polynary immunoassay board - Google Patents
Polynary immunoassay board Download PDFInfo
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- CN1399132A CN1399132A CN 02112229 CN02112229A CN1399132A CN 1399132 A CN1399132 A CN 1399132A CN 02112229 CN02112229 CN 02112229 CN 02112229 A CN02112229 A CN 02112229A CN 1399132 A CN1399132 A CN 1399132A
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- orifice plate
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- immunoassay
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Abstract
The invented polynary immunoassay plate relates to an enzyme immunoassay chip reaction device which is applied widely to the medical detection, analysis and other field. The polynary immuoassay plate has several holes with antigen fixed by chemical bond to an orifice plate, the surface of the orifice plate as solid carrier is closed and the sample to be tested is added to produce hybridized reaction with the biomolecules fixed to the orifice plate. After unreacted matter is washed off, the labeled antigen is added for hybridization, and the unreacted matter is washed off again before the label signal is detected. Several indexes may be detected simultaneously. The present invention is flexible and saving in tested sample amount, and some commercial autoamtic instruments may be used.
Description
Technical field
Polynary immunoassay board of the present invention relates to the chip reaction unit in a kind of enzyme immunoassay (EIA), is widely used in fields such as medical treatment detection, analysis.
Background technology
Engvall in 1971 and Perlmann have delivered Enzyme Linked Immunoadsorbent Assay mensuration (enzyme-linked immunosorbent assay, ELISA) be used for the article of IgG quantitative measurement, make the enzyme labelled antibody technical development that began to be used for Antigen Location in 1966 become the assay method of liquid sample micro substance.The ultimate principle of this method is: 1. make antigen or antibodies to certain surface of solid phase carriers, and keep its immunocompetence.2. make antigen or antibody and certain enzyme connect into enzyme-labelled antigen or antibody, this enzyme-labelled antigen or antibody had both kept its immunocompetence, kept the activity of enzyme again.When measuring, reacted examining sample (mensuration antibody or antigen wherein) and enzyme-labelled antigen or antibody antigen or antibody by different steps and surface of solid phase carriers.Method with washing makes the antigen antibody complex that forms on the solid phase carrier separate with other materials, and the amount that is combined in examined object matter in enzyme amount and the sample on the solid phase carrier at last becomes certain ratio.After adding the substrate of enzyme reaction, substrate is become coloured product by enzymatic, and the amount of product is directly related with the amount of examined object matter in the sample, so can carry out qualitative or quantitative test according to the depth of color reaction.Because the catalysis frequency of enzyme is very high, thus reaction effect can greatly be amplified, thus make assay method reach very high susceptibility.
Enzyme immunoassay (EIA) has the susceptibility and the specificity of height, and nearly all soluble antigen antibody forming system all can be in order to detect.Its I measured value reaches ng even pg level.Compare with radiommunoassay, the advantage of enzyme immunoassay (EIA) is that labelled reagent is more stable, and "dead" harm.Therefore, the application of enzyme immunoassay (EIA) is maked rapid progress, and new method, the new technology of enzyme immunoassay (EIA) constantly develop.ELISA is widely used, and can comprise the following aspects in order to the project that detects: 1. pathogen and antibody thereof: be widely used in the diagnosis of infectious disease.2. protein: various immunoglobulin (Ig)s, complement component, tumor markers, various plasma proteinss, isodynamic enzyme, hormone.3. non-peptide hormone: as T3, T4, estrogen, cortisol etc.4. medicine and drugs: as digoxin, phenobarbital, gentamicin, morphine etc.
But the universal appearance that should give the credit to commercially available reagent box and automatic or semi-automatic detecting instrument of enzyme immunoassay (EIA) in medical test.Comprise bag by good solid phase carrier, enzyme conjugates substrate and cleansing solution etc. in the commodity ELISA kit.The essential outstanding instrument (special-purpose be called ELISA plate reading), wash plate is also extremely useful in quantitative measurement for the ELISA all appts.The use of rinsing maching is the time and labour saving not only, and is beneficial to operational standardization, and centering small test chamber is practical and is easy to accept.It is ripe that semi-automatic and robotization elisa assay instrument also becomes, and obtain application in big-and-middle-sized Clinical Test Lab.The former is applicable to that the ELISA of 96 all orifice plates measures; The latter only with the supporting use of particular agent.
Though ELISA has the susceptibility and the specificity of height, is widely used, and is of a great variety, only wrap by a kind of antigen or antibody in its every hole of 96 orifice plate of commercialization ELISA kit of using now, so a kind of kit can only detect a kind of antigen or antibody.Detect several different indexs as need, then need to repeat much at one test procedure with different kits, the wasting manpower and material resources has also prolonged the time of diagnosis.
Summary of the invention
The objective of the invention is to: a kind of polynary immunoassay board is provided, and promptly a kind of 96 new AND DEWATERING FOR ORIFICE STRUCTURE make it can wrap by several different antigens (or antibody) in every hole, thereby make experiment more time saving and energy saving.
Purpose of the present invention can be achieved through the following technical solutions: a kind of polynary immunoassay board, be with antigen (or antibody) dot matrix in each hole of orifice plate, antigen (or antibody) is connected on the orifice plate by chemical bond-linking and is fixed, after the orifice surface process sealing as solid phase carrier, add testing sample, carry out hybridization reaction with biomolecule fixing on the orifice plate, behind the unconjugated material of flush away, two anti-(or antigens) that the adding mark is crossed are hybridized certification mark signal behind the unconjugated material of flush away again.The power of detected marking signal becomes certain positive correlation with the content of composition to be measured in the sample.Its principle detects the antigen schematic diagram referring to Fig. 1 polynary immunoassay board, and Fig. 2 polynary immunoassay board detects shown in the antibody schematic diagram.
Wherein, described orifice plate is 96 orifice plates, and the bottom of this 96 orifice plate can be that glass sheet, acetate film, cellulose nitrate film, nylon membrane, silicon chip, steel disc, potsherd, plastic sheet and bottom that polystyrene plastics or surface have an activated group are posted or had a kind of of acetate film, cellulose nitrate film, nylon membrane, silicon chip, steel disc, potsherd, plastic sheet etc.
Testing sample can be one or more in the cell pyrolysis liquid of serum, body fluid, tissue fluid, tissue.
Described two labels anti-or antigen can be fluorescent dye or chemiluminescent substance.
The preparation method of polynary immunoassay board of the present invention comprises: the first, and the point sample method: (1) uses the high speed spotting robot, determines the arrangement mode and the point sample position of dot matrix according to the kind of antigen; (2) direct dot matrix was in the bottom of 96 orifice plates after spotting needle took out sample from porous plate; (3) ambient temperature overnight or 37 ℃ of incubations are 1 hour, with fixed sample; (4) add confining liquid at carrier surface, 37 ℃ of incubations 1 hour; (5) fully wash and drying at room temperature with damping fluid; Second step, two anti-marks: anti-with fluorescent dye or chemiluminescent substance mark two, remove not binding constituents after, 4 ℃ of preservations are standby.
Hybridization reaction: sample adds the excellent 96 orifice plate reacting holes of point, and 37 ℃ of incubations make material to be detected be incorporated into the solid phase bottom after about 30 minutes, drips mark two anti-or antigens again, 37 ℃ of lucifuge incubations 30 minutes.Clean and room temperature is dried.
At last, result's detection: after reaction finishes, utilize the reading analysis system of specialty to carry out result's interpretation.
Characteristics of the present invention, advantage, result of use: the first, many indexs detect simultaneously, and this 96 orifice plate is because with in several different antigen (or antibody) dot matrix each holes on solid phase carrier 96 orifice plates, thereby can detect several different indexs simultaneously in every hole.Repetitive operation is loaded down with trivial details when having abandoned ELISA in the past and detecting several different index, has saved manpower and materials, has also shortened the time of diagnosis.The second, test sample neatly.Utilize 96 orifice plates dismountings, assembly unit flexibly, can be random detecting at any time as required and diagnosing obtains the result with the fastest speed, for patient and doctor strive for valuable treatment time.The 3rd, the amount of more saving testing sample.Owing to can carry out the detection of several different indexs simultaneously, need not detect several times, thereby save the amount of testing sample greatly with several different kits.The 4th, can save a large amount of funds.Because of the present invention's 96 orifice plate outward appearances are identical with 96 former orifice plates, the commercial automatic or semi-automatic instrument and equipment of enzyme immunoassay (EIA) such as rinsing maching, the full-automatic consersion unit of ELISA etc. ripe and that popularize can be used, so need not purchase instrument again.
Description of drawings
Accompanying drawing 1, polynary immunoassay board detects the antigen schematic diagram.
Accompanying drawing 2, polynary immunoassay board detects the antibody schematic diagram.
Accompanying drawing 3 carries out the detection synoptic diagram of multiple disease simultaneously.
Embodiment:
Shown in the detection synoptic diagram that Fig. 3 carries out multiple disease simultaneously, with the fluorescently-labeled two anti-hepatitis antibodies that detect in the serum.Sample application array is 4 * 4.Left side figure is that antigen dot matrix way A1-A4 is a HA antigen, and B1-B4 is a hepatitis B surface antibody, and C1-C4 is a hepatitis C antigen, and D1-D4 is a HDAg; Right figure detects the hepatitis antibody result, and the result is shown as the hepatitis B surface antibody positive, the antibody to hepatitis D positive, HAAb and antibody to hepatitis C feminine gender, and the power of fluorescence is represented the height of antibody content.
Claims (6)
1, a kind of polynary immunoassay board, be with antigen (or antibody) dot matrix in each hole of orifice plate, be characterised in that: antigen (or antibody) is connected on the orifice plate by chemical bond-linking and is fixed, and after the orifice surface process sealing as solid phase carrier, adds testing sample, carry out hybridization reaction with biomolecule fixing on the orifice plate, behind the unconjugated material of flush away, two anti-(or antigens) that the adding mark is crossed are hybridized certification mark signal behind the unconjugated material of flush away again.
2, polynary immunoassay board according to claim 1, be characterised in that described orifice plate is 96 orifice plates, the bottom of this 96 orifice plate can be that glass sheet, acetate film, cellulose nitrate film, nylon membrane, silicon chip, steel disc, potsherd, plastic sheet and bottom that polystyrene plastics or surface have an activated group are posted or had a kind of of acetate film, cellulose nitrate film, nylon membrane, silicon chip, steel disc, potsherd, plastic sheet etc.
3, according to claim 1,2 described polynary immunoassay boards, be characterised in that described testing sample can be one or more in the cell pyrolysis liquid of serum, body fluid, tissue fluid, tissue.
4, polynary immunoassay board according to claim 3 is characterised in that described two labels anti-or antigen can be fluorescent dye or chemiluminescent substance.
5, the preparation method of polynary immunoassay board according to claim 3, be characterised in that the first point sample method: (1) uses the high speed spotting robot, determines the arrangement mode and the point sample position of dot matrix according to the kind of antigen; (2) direct dot matrix was in the bottom of 96 orifice plates after spotting needle took out sample from porous plate; (3) ambient temperature overnight or 37 ℃ of incubations are 1 hour, with fixed sample; (4) add confining liquid at carrier surface, 37 ℃ of incubations 1 hour; (5) fully wash and drying at room temperature with damping fluid; Second step, two anti-marks: anti-with fluorescent dye or chemiluminescent substance mark two, remove not binding constituents after, 4 ℃ of preservations are standby.
6, the preparation method of polynary immunoassay board according to claim 5, be characterised in that its hybridization reaction is that sample is added the excellent 96 orifice plate reacting holes of point, about 30 minutes of 37 ℃ of incubations, make material to be detected be incorporated into solid phase bottom, drip mark two anti-or antigens again, 37 ℃ of lucifuge incubations 30 minutes, after clean and room temperature is dried, utilize the reading analysis system of specialty to carry out result's interpretation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02112229 CN1399132A (en) | 2002-06-25 | 2002-06-25 | Polynary immunoassay board |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02112229 CN1399132A (en) | 2002-06-25 | 2002-06-25 | Polynary immunoassay board |
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| Publication Number | Publication Date |
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| CN1399132A true CN1399132A (en) | 2003-02-26 |
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| CN 02112229 Pending CN1399132A (en) | 2002-06-25 | 2002-06-25 | Polynary immunoassay board |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1845778B (en) * | 2003-09-03 | 2010-09-15 | Cedi诊断私人有限公司 | Method of detecting multiple analytes |
| CN101988921A (en) * | 2009-07-29 | 2011-03-23 | 丹耐克斯技术有限公司 | Sample plate |
| CN102207504A (en) * | 2011-03-23 | 2011-10-05 | 北京华创远航科技有限公司 | Enzyme-linked immunosorbent assay kit, and preparation method thereof |
| CN104280556A (en) * | 2014-10-31 | 2015-01-14 | 杨子学 | Detection method and kit for simultaneously determining content of lipoprotein phospholipase A2 and C reactive protein in blood plasma |
| CN105652000A (en) * | 2014-12-31 | 2016-06-08 | 天津东亚生物技术有限公司 | Novel protein chip |
| US9523701B2 (en) | 2009-07-29 | 2016-12-20 | Dynex Technologies, Inc. | Sample plate systems and methods |
-
2002
- 2002-06-25 CN CN 02112229 patent/CN1399132A/en active Pending
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1845778B (en) * | 2003-09-03 | 2010-09-15 | Cedi诊断私人有限公司 | Method of detecting multiple analytes |
| CN101988921A (en) * | 2009-07-29 | 2011-03-23 | 丹耐克斯技术有限公司 | Sample plate |
| CN101988921B (en) * | 2009-07-29 | 2014-05-14 | 丹耐克斯技术有限公司 | Sample plate |
| US9244069B2 (en) | 2009-07-29 | 2016-01-26 | Dynex Technologies | Sample plate systems and methods |
| US9523701B2 (en) | 2009-07-29 | 2016-12-20 | Dynex Technologies, Inc. | Sample plate systems and methods |
| US9857367B2 (en) | 2009-07-29 | 2018-01-02 | Dynex Technologies, Inc. | Sample plate systems and methods |
| US10207268B2 (en) | 2009-07-29 | 2019-02-19 | Dynex Technologies, Inc. | Sample plate systems and methods |
| US10969386B2 (en) | 2009-07-29 | 2021-04-06 | Dynex Technologies, Inc. | Sample plate systems and methods |
| CN102207504A (en) * | 2011-03-23 | 2011-10-05 | 北京华创远航科技有限公司 | Enzyme-linked immunosorbent assay kit, and preparation method thereof |
| CN104280556A (en) * | 2014-10-31 | 2015-01-14 | 杨子学 | Detection method and kit for simultaneously determining content of lipoprotein phospholipase A2 and C reactive protein in blood plasma |
| CN105652000A (en) * | 2014-12-31 | 2016-06-08 | 天津东亚生物技术有限公司 | Novel protein chip |
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