CN1067512A - Immunoglobulin G AM associating rocket electrophoresis assay plate and assay method - Google Patents
Immunoglobulin G AM associating rocket electrophoresis assay plate and assay method Download PDFInfo
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- CN1067512A CN1067512A CN 92103656 CN92103656A CN1067512A CN 1067512 A CN1067512 A CN 1067512A CN 92103656 CN92103656 CN 92103656 CN 92103656 A CN92103656 A CN 92103656A CN 1067512 A CN1067512 A CN 1067512A
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Abstract
The invention provides a kind of IGAM associating rocket electrophoresis assay plate and assay method, this assay plate is characterised in that anti-human immune globulin serum contained in the agarose-gel film is the two or more potpourri that is selected from anti-immunoglobulin G while, A, M serum.Can the simultaneous determination IGAM with this assay plate, promptly add a sample and can obtain three results simultaneously with a plate, compare with an existing plate individual event determination method, starting material and running time are reduced greatly, improved detection efficiency, be suitable for laboratories at different levels and carry out mass detection.
Description
The present invention relates to a kind of immunoglobulin (Ig) rocket electrophoresis assay plate and assay method, particularly a kind of IGAM associating rocket electrophoresis assay plate and assay method.
Immunoglobulin (Ig) rocket electrophoresis determination method be make antigen under the electric field force effect when containing the agarose gel plate of antibody and corresponding antibody form antigen antibody complex, this compound precipitates at antigen and the well-proportioned position of antibody and forms conical precipitation peak, its shape is like rocket, because the height at precipitation peak is directly proportional with antigen concentration, so it is quantitative to can be used as antigen.
In the prior art, immunoglobulin G, A, M rocket electrophoresis assay method is that a plate individual event is measured, only contain a kind of anti-human immune globulin serum (promptly anti-immunoglobulin G while in the agarose-gel film of assay plate, A, a kind of in the M serum), therefore a plate once can only be measured a result, if will measure immunoglobulin G in the tested serum, A, the content of M then needs three assay plate to divide mensuration three times, each running time of this in addition rocket electrophoresis needs several hours, therefore a this plate individual event determination method is not suitable for mass detection, causes this precision height, good reproducibility, reagent source detection method easily is difficult to promote the use of in enormous quantities.(reference: " clinical immunoassay test " p275～276, Tianjin science tech publishing house publishes nineteen ninety)
The objective of the invention is to overcome above-mentioned the deficiencies in the prior art part, a kind of rocket electrophoresis assay plate and assay method of energy simultaneous determination IGAM is provided.
The object of the present invention is achieved like this:
This IGAM associating rocket electrophoresis assay plate is to be cast with the agarose-gel film that one deck contains anti-human immune globulin serum on support plate, be equipped with the aperture of be used to annotate immunoglobulin (Ig) working stamndard and tested serum on the film, contained anti-human immune globulin serum is the two or more potpourri that is selected from anti-immunoglobulin G while, A, M serum in the gel mould.
Said mixture is meant optional the two potpourri among anti-immunoglobulin G while, A, M serum three's potpourri or the three.
Anti-human immune globulin serum's consumption (in every ml Ago-Gel) contained in the said determination plate gel mould is respectively (" u " is " unit "):
Anti-immunoglobulin G while serum 0.1～0.5u is preferably 0.2～0.3u;
Anti-human immunoglobulin A's serum 0.1～1u is preferably 0.4～0.6u;
Anti-human immunoglobulin M's serum 0.1～1u is preferably 0.4～0.6u;
The determining of above-mentioned anti-human immune globulin serum's consumption be based on (1) for the precipitation peak that makes assay plate and behind electrophoresis, occur at 2～4cm height, prevent the too dense top that do not reach within a certain period of time of antigen, and antibody is too dense measured too low being difficult to of precipitation peak; (2) for the peak shape (specifically introducing in the embodiment of back) that makes the IGAM precipitation peak of assay plate through occurring behind the electrophoresis remains unchanged in the ordinary course of things, so that confirm.
With the method for this assay plate simultaneous determination IGAM, comprise be filled in the aperture of assay plate gel mould respectively with immunoglobulin (Ig) working stamndard after the diluted and tested serum, bridging electrophoresis, dyeing and rinsing.Used dilution is 1 part of 2.5～3% formaldehyde (AR level) aqueous solution and the potpourri of 5 parts of 7.5～8.5mol/L aqueous solution of urea.
The effect of formaldehyde is the precipitation peak clear display that makes immunoglobulin A, M in the above-mentioned dilution, and the effect of urea is the precipitation peak clear display that makes immunoglobulin G.
The present invention contrasts that prior art has the following advantages and good effect:
IGAM associating rocket electrophoresis assay plate provided by the invention and assay method not only have the advantage (precision height, good reproducibility, reagent source convenience etc.) that prior art one plate individual event is measured, and owing to add a sample with a plate and can obtain three results simultaneously, and its measurement result is consistent with the result that a plate individual event is measured, therefore starting material and running time are reduced greatly, improved detection efficiency, be suitable for laboratories at different levels and carry out mass detection.
Below in conjunction with specific embodiment describe the present invention (following " immunoglobulin (Ig) " is abbreviated as " Ig "):
Making sheet: support plate is 10 * 7cm plastic plate (also can be glass plate), every plate 10g/L Ago-Gel 11ml, anti-human IgG serum (tiring 1: 100) 2.52u, anti-people IgA serum (1:90 tires) 3.18u, the anti-human IgG of anti-people IgM serum (1:120 tires) 5.12u(, A, M serum is the goods of Beijing Biological Product Inst., Ministry of Public Health), with above-mentioned anti-human IgG, A, M serum and Ago-Gel are in being cast in behind 55 ℃ of mixings on the support plate of horizontal positioned, the cooling back is along bottom (long limit) punching one row of gel mould on the support plate, aperture 2.5mm, pitch-row 2.0mm.
Measure:
Get 1 part of 2.85% formaldehyde (AR level) aqueous solution and mix for 5 parts, make dilution with the 8.0mol/L aqueous solution of urea;
With above-mentioned dilution tested serum was diluted by 1: 10, (the Ig working stamndard was the goods of Beijing Biological Product Inst., Ministry of Public Health by 1: 10,1: 20,1: 30 three kinds of concentration dilutions respectively with the Ig working stamndard with above-mentioned dilution, wherein Ig content is: IgG126u/ml, 80.4 μ g/u; IgA106u/ml, 14.2 μ g/u; IgM128u/ml, 8.47 μ g/u);
The tested serum 5 μ ml that get after the above-mentioned dilution are filled in the aperture of assay plate gel mould, getting above-mentioned each 5 μ ml of Ig working stamndard that are diluted to three kinds of concentration is filled into respectively in three apertures, put up a bridge with four layers of gauze then, place in PH=9.0, the 0.05M barbitol buffer solution electrophoresis tank and carried out electrophoresis 4.5 hours in electric current 20～25mA/ plate, terminal voltage 5～7V;
Be placed in the 5% glacial acetic acid rinsing liquid rinsing 10 minutes in 3 minutes to the peak shape clear display with amino black 10B dye liquor overlay face.
Can measure 20 parts of tested serum with the every plate of the assay plate of present embodiment.
Peak shape is judged: assay plate is behind electrophoresis, upwards move the variform rocket shapes precipitation peaks that three of appearance are superimposed along each aperture, the peak shape that occurs when peak shape is measured with a plate individual event is identical, that is: the peak base is the wideest, peak height is the highest, the peak sharp the sharpest be IgG precipitation peak, peak base, peak height are placed in the middle, sharp what be blunt round is IgA precipitation peak at the peak, and the peak base is the narrowest, peak height is minimum, sharp to be more pointed be IgM precipitation peak at the peak.
IgG, the A, the M that accurately measure various concentration Ig working stamndards formation precipitate peak heights, the drawing standard curve, and IgG, the A, the M that measure tested serum formation again precipitate peak heights, through consulting typical curve, can draw the content of IgG, A, M in the tested serum.
It is as follows that assay plate provided by the invention and assay method are carried out linear relationship, replica test and correlation test interpretation of result:
1. linear relationship: the Ig working stamndard drawing standard curve that will be diluted to 1: 5,1: 10,1: 15,1: 20,1: 25,1: 30 several variable concentrations, its as a result IgG content in 3.38～20.26g/L scope with the peak height relation of being in line, r=0.9959, a=3.9447, b=7.0886; IgA content in 0.5～3.01g/L scope with the peak height relation of being in line, r=0.9996, a=6.8733, b=1.3772; IgM content in 0.36～2.1g/L scope with the peak height relation of being in line, r=0.9994, a=1.7883, b=20.8214.
2. replica test analysis: do 10 times in a tested serum specimen is criticized, did in one day between batch and once do altogether 10 times, IgG content is 10.071g/L as a result, CV=0.77% in crowd, CV=1.3% in crowd; IgA content is 1.498g/L, CV=0.70% in crowd, CV=1.5% between crowd; IgM content is 1.0784g/L, CV=0.46% in crowd, CV=1.2% between crowd.The repeatability that proves this assay method is good.
3. correlation test analysis: do correlativity relatively with machine testing 48 routine clinical samples, its result with an assay method provided by the invention and an existing plate individual event assay method:
IgG r=0.9430
(this law)=0.8686X+1.3916
IgA r=0.9440
(this law)=0.8935X+0.138
IgM r=0.9917
(this law)=1.0534X+0.0856
Three P＞0.05 proves the measurement result there was no significant difference of three measurement results of a plate provided by the invention and a plate individual event.
Claims (3)
1, a kind of immunoglobulin (Ig) rocket electrophoresis assay plate, particularly a kind of IGAM associating rocket electrophoresis assay plate, on support plate, be cast with one deck and contain anti-human immune globulin serum's agarose-gel film, be equipped with the aperture of be used to annotate immunoglobulin (Ig) working stamndard and tested serum on the film, it is characterized in that anti-human immune globulin serum contained in the gel mould is the two or more potpourri that is selected from anti-immunoglobulin G while, A, M serum.
2, assay plate as claimed in claim 1 is characterized in that anti-human immune globulin serum's consumption contained in the gel mould (in every ml Ago-Gel) is respectively (" u " is " unit "):
Anti-immunoglobulin G while serum 0.1～0.5u is preferably 0.2～0.3u;
Anti-human immunoglobulin A's serum 0.1～1u is preferably 0.4～0.6u;
Anti-human immunoglobulin M's serum 0.1～1u is preferably 0.4～0.6u;
3, a kind of method with the described assay plate simultaneous determination of claim 1 IGAM, comprise will be filled in the aperture of assay plate gel mould respectively with immunoglobulin (Ig) working stamndard after the diluted and tested serum, bridging electrophoresis, dyeing and rinsing, it is characterized in that used dilution is 1 part of 2.5～3% formaldehyde (AR level) aqueous solution and the potpourri of 5 parts of 7.5～8.5mol/L aqueous solution of urea.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 92103656 CN1067512A (en) | 1992-05-14 | 1992-05-14 | Immunoglobulin G AM associating rocket electrophoresis assay plate and assay method |
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| Application Number | Priority Date | Filing Date | Title |
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| CN 92103656 CN1067512A (en) | 1992-05-14 | 1992-05-14 | Immunoglobulin G AM associating rocket electrophoresis assay plate and assay method |
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| CN1067512A true CN1067512A (en) | 1992-12-30 |
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| CN 92103656 Pending CN1067512A (en) | 1992-05-14 | 1992-05-14 | Immunoglobulin G AM associating rocket electrophoresis assay plate and assay method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1890566B (en) * | 2003-12-26 | 2011-07-20 | 松下电器产业株式会社 | Biological sample discriminating device, biological sample discriminating method, and biological sample discriminating plate |
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1992
- 1992-05-14 CN CN 92103656 patent/CN1067512A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1890566B (en) * | 2003-12-26 | 2011-07-20 | 松下电器产业株式会社 | Biological sample discriminating device, biological sample discriminating method, and biological sample discriminating plate |
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