CN1412557A - ToRCH protein chip and its preparation method - Google Patents
ToRCH protein chip and its preparation method Download PDFInfo
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- CN1412557A CN1412557A CN 01134030 CN01134030A CN1412557A CN 1412557 A CN1412557 A CN 1412557A CN 01134030 CN01134030 CN 01134030 CN 01134030 A CN01134030 A CN 01134030A CN 1412557 A CN1412557 A CN 1412557A
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Abstract
The present invention discloses a ToRCH protein chip. Said chip includes glass slide, the high molecular polysaccharide film is adsorbed on the slide, and the ToRCH antigen-antibody protein microarray is adsorbed on the surface of the high molecular polysaccharide film. Said invention method includes the following steps: cleaning glass slide, dissolving high-molecular polysacchairde raw material ins olvent; adding above-mentioned dissolved material on the glass slide, gel-dewatering and oxidation and dropping ToRCH antigen-antibody microarray to implement preparation of chip. Said invention can use one chip to implement all the ToRCH diagnosises by one-step operation within about 2 hr.
Description
Technical field
The present invention relates to a kind of protein-chip, especially a kind of protein-chip of diagnosing usefulness the invention still further relates to its preparation method simultaneously, belongs to the biochip technology field.
Background technology
Nineteen nineties, a brand-new era is brought biotechnology in biochip research at the very start.Compare with the genetic chip that causes extensive concern, protein-chip can reflect the protein level in the biosome, therefore can express more complete biological information.
" ToRCH " is the technical term of modern perinatal medicine.Wherein, T represents toxoplasma gondii (Toxoplasmagondii), O represents (Other) such as Epstein-Barr virus, HIV virus and human parvovirus B19s, R represents rubella virus (Rubella Virus), C represents cytomegalovirus (Cytomegalovirus), and H represents herpes simplex I type and II type virus (Herpes Simplex Virus I and II).The scope of these pathogenic infections is wide, harm is big, and the pregnant woman is easy to take place primary infection, causes miscarriage, premature labor, stillborn foetus or growth retardation of fetus and developmental deformity, thus the population quality of having a strong impact on.
Developed countries such as America and Europe just classify it as pregnancy period examination project as far back as the seventies.Pregnant earlier T oRCH testing, especially Beijing have also progressively been carried out in China developed regions, and municipal government has issued the relevant document of ToRCH being classified as pregnant woman's routine inspection project.
At present, the complete dependence on import of kit of China ToRCH deagnostic test, it costs an arm and a leg, because the every box reagent of kit need use up in the short as far as possible time, therefore requires the number of inspection simultaneously and the dosage coupling of every box reagent.The person under inspection need spend 300-500 unit, usually will wait for that there is the result in several old name for the Arabian countries in the Middle Easts.Expensive inspection charge and loaded down with trivial details scrutiny program have limited popularizing that ToRCH checks to a great extent.
The objective of the invention is to: at the problem of above-mentioned existing ToRCH deagnostic test reagent existence, utilize the high-tech biotechnology to propose a kind of ToRCH protein-chip, also will provide the preparation method of this ToRCH protein-chip simultaneously, so that the diagnosis of ToRCH disease has more science, checks diagnostic method easily, both can make things convenient for hospital, the patient generaI investigation of also taking like a shot.
In order to reach the foregoing invention purpose, ToRCH protein-chip of the present invention comprises microslide, and described microslide is attached with macromolecule glycan film, and described macromolecule glycan film surface attachment has ToRCH antigen-antibody protein microarray.
The preparation method of ToRCH protein-chip of the present invention is as follows: A. cleans microslide, cleans the back airing; B. macromolecule glycan pleurodiaphragmatic in terspace material is dissolved in the solvent, concentration is controlled at 1%～10%; C. again this solution is added on the microslide behind the airing cooled and solidified; D. the microslide of cooled and solidified is heated, make gel slough moisture, form film; E. add the sodium periodate that concentration is 1～100 mM, make film oxidation; F. on the film after the oxidation with a ToRCH antigen microarray; H. point adds ToRCH antibody on the ToRCH antigen microarray, finishes the preparation of chip.
Above step at first makes the macromolecule glycan film polymerization in the process of heating that is coated on the microslide fix and slough moisture; Then under the oxidation of sodium periodate, the sugar shell generates aldehyde radical, and when aldehyde radical contacted with ToRCH proteantigen antibody, the amino of aldehyde radical and protein rare husband was taken place reacts (Schiff ' s), generate the covalent bonds of efficiently tiring, the ToRCH antigen-antibody is securely fixed on the chip.
During inspection, be coated on clearly on the chip of the present invention as long as will wait to have a blood test, if contain ToRCH virus in the serum, with the corresponding antigen-antibody protein on very fast and the chip take place special should, when by fluorescence microscope, just show corresponding positive feature, like this in the short time about two hours, just can finish all ToRCH diagnosis by a slice chip is disposable, very convenient quick, its cost is significantly less than the expense that the import reagent box is checked.Therefore, after the present invention's release, will greatly promote popularizing of ToRCH disease examination diagnosis, to the prenatal and postnatal care and the important and far-reaching meaning of raising newborn population quality generation in the China and even the world.
Description of drawings
The invention will be further described below in conjunction with accompanying drawing.Fig. 1 is the structural representation of the embodiment of the invention one.
Embodiment
Embodiment one
The ToRCH protein-chip of present embodiment adopts glass as microslide 1, the upper surface of microslide 1 is attached with one deck agarose film 2, the upper surface point of agarose film 2 has ToRCH antigen protein microarray 3, and point has ToRCH antibody 4 (referring to Fig. 1) on the ToRCH antigen microarray 3.
The ToRCH protein-chip of present embodiment adopts the following steps preparation: A. cleans glass slide with the soda acid cleaning fluid, and is clean with pure water rinsing again, then airing; B. add agarose in water, heating dissolves, and concentration is controlled at 3%～5%; C. this solution is added on the microslide behind the airing, natural cooling solidifies; D. the microslide of cooled and solidified is put into baking oven or exsiccator is heated to 45～50 ℃, make gel slough moisture, form film; E. add the sodium periodate that concentration is 20 mMs, make film oxidation; F. on the film after the oxidation, put with the ToRCH antigen microarray; G. point adds ToRCH antibody on the ToRCH antigen microarray, finishes the preparation of chip.
Embodiment two
The ToRCH protein-chip of present embodiment adopts silicon chip as microslide 1, the upper surface of microslide 1 is attached with one deck glucosan---polyacrylamide film 2, the upper surface point of glucosan---polyacrylamide film 2 has ToRCH antigen protein microarray 3, and point has ToRCH antibody 4 (same Fig. 1) on the ToRCH antigen microarray 3.
The ToRCH protein-chip of present embodiment adopts following steps preparation: A. with soda acid cleaning fluid cleaning silicon chip microslide, and is clean with pure water rinsing again, then airing; B. agarose is dissolved in the polyacrylamide ammonia solution, concentration is controlled at 6%～8%, and adds the polymerizer 10 μ l% of 10 millimolar concentrations; C. this solution is added on the microslide behind the airing, after illumination, makes its polymeric gel again; D. the microslide of cooled and solidified is put into baking oven or exsiccator is heated to 45～50 ℃, make gel slough moisture, form film; E. add the sodium periodate that concentration is 20 mMs, make film oxidation; F. point adds the ToRCH antigen microarray on the film after the oxidation; G. the ToRCH antibody of adding some points on the ToRCH antigen microarray is finished the preparation of chip.
During inspection, as long as on the microslide behind the airing that is coated on the foregoing description clearly of will waiting to have a blood test, in the short time about two hours, just can finish all ToRCH diagnosis by a slice chip is disposable, very convenient quick.Experiment showed, that its sensitivity and specificity are all very good, its cost only is 30% of an import reagent box inspection charge.Therefore, after the release, will have very vast market prospect, will have a tremendous social and economic benefits.
In addition to the implementation, the present invention also has other numerous embodiments.For example, microslide adopts Ge, GaAs, GaP, SiO2, Si3N4 sheet, or gel, glycan thing such as poly-tetrafluoro, poly-alum, polycarbonate; Macromolecule glycan film is an agarose---polyacrylamide film or agarose---glucosan---polyacrylamide film etc.; All employings are equal to the technical scheme of replacement or equivalent transformation formation, are the protection domain of requirement of the present invention.
Claims (10)
1. a ToRCH protein-chip comprises microslide, it is characterized in that: described microslide is attached with macromolecule glycan film, and described macromolecule glycan film surface attachment has ToRCH antigen-antibody protein microarray.
2. ToRCH protein-chip according to claim 1 is characterized in that: described macromolecule glycan film upper surface point has ToRCH antigen protein microarray, and point has ToRCH antibody on the ToRCH antigen microarray.
3. ToRCH protein-chip according to claim 1 and 2 is characterized in that: described microslide is a glass, and the upper surface of described glass slide is attached with one deck agarose film.
4. ToRCH protein-chip according to claim 1 and 2 is characterized in that: described microslide is a silicon chip, and the upper surface of silicon chip microslide is attached with one deck glucosan---the polyacrylamide film.
5. ToRCH protein-chip according to claim 1 is characterized in that: described macromolecule glycan film is an agarose---polyacrylamide film or agarose---glucosan---polyacrylamide film.
6. ToRCH protein-chip preparation method according to claim 1 is characterized in that may further comprise the steps: A. cleans microslide, cleans the back airing; B. macromolecule glycan pleurodiaphragmatic in terspace material is dissolved in the solvent, concentration is controlled at 1%～10%; C. again this solution is added on the microslide behind the airing cooled and solidified; D. the microslide of cooled and solidified is heated, make gel slough moisture, form film; E. add the sodium periodate that concentration is 1～100 mM, make film oxidation; F. point adds the ToRCH antigen microarray on the film after the oxidation; G. point adds ToRCH antibody on the ToRCH antigen microarray.
7. ToRCH protein-chip preparation method according to claim 6 is characterized in that may further comprise the steps: A. cleans glass slide with the soda acid cleaning fluid, and is clean with pure water rinsing again, then airing; B. add agarose in water, heating dissolves, and concentration is controlled at 3%～5%; C. this solution is added on the microslide behind the airing, natural cooling solidifies; D. the microslide of cooled and solidified is put into baking oven or exsiccator is heated to 45～50 ℃, make gel slough moisture, form film; E. add the sodium periodate that concentration is 20 mMs, make film oxidation; F. point adds the ToRCH antigen microarray on the film after the oxidation; G. point adds ToRCH antibody on the ToRCH antigen microarray.
8. ToRCH protein-chip preparation method according to claim 6 is characterized in that may further comprise the steps: A. is with soda acid cleaning fluid cleaning silicon chip microslide, and is clean with pure water rinsing again, then airing; B. agarose is dissolved in the polyacrylamide ammonia solution, concentration is controlled at 6%～8%, and adds the polymerizer 10 μ l% of 10 millimolar concentrations; C. this solution is added on the microslide behind the airing, makes its polymeric gel again after illumination; D. the microslide of cooled and solidified is put into baking oven or exsiccator is heated to 45～50 ℃, makes gel slough moisture, forms film; E. add the sodium periodate that concentration is 20 mMs, make film oxidation; F. point adds the ToRCH antigen microarray on the film after the oxidation; G. point adds ToRCH antibody on the ToRCH antigen microarray.
9. ToRCH protein-chip preparation method according to claim 6 is characterized in that: described macromolecule glycan is agarose a---polyacrylamide.
10. ToRCH protein-chip preparation method according to claim 6 is characterized in that: described macromolecule glycan is agarose---glucosan---polyacrylamide.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01134030 CN1412557A (en) | 2001-10-11 | 2001-10-11 | ToRCH protein chip and its preparation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 01134030 CN1412557A (en) | 2001-10-11 | 2001-10-11 | ToRCH protein chip and its preparation method |
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| CN1412557A true CN1412557A (en) | 2003-04-23 |
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| CN 01134030 Pending CN1412557A (en) | 2001-10-11 | 2001-10-11 | ToRCH protein chip and its preparation method |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100334229C (en) * | 2005-06-17 | 2007-08-29 | 东南大学 | Preparation method of DNA microarray chip based on gel fixed nucleic acid |
| CN101833001A (en) * | 2010-04-21 | 2010-09-15 | 黄若磐 | Protein chip kit for detecting inflammatory factors and preparation method thereof |
| CN106093427A (en) * | 2016-06-02 | 2016-11-09 | 滨州医学院 | Identify the new method of toxoplasma, rubella virus and herpes simplex virus simultaneously |
| WO2019169792A1 (en) * | 2018-03-07 | 2019-09-12 | 深圳市伯劳特生物制品有限公司 | Anti-torch-igm antibody repertoire chip, preparation method therefor and torch detection kit |
-
2001
- 2001-10-11 CN CN 01134030 patent/CN1412557A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100334229C (en) * | 2005-06-17 | 2007-08-29 | 东南大学 | Preparation method of DNA microarray chip based on gel fixed nucleic acid |
| CN101833001A (en) * | 2010-04-21 | 2010-09-15 | 黄若磐 | Protein chip kit for detecting inflammatory factors and preparation method thereof |
| CN101833001B (en) * | 2010-04-21 | 2013-08-28 | 广州瑞博奥生物科技有限公司 | Protein chip kit for detecting inflammatory factors and preparation method thereof |
| CN106093427A (en) * | 2016-06-02 | 2016-11-09 | 滨州医学院 | Identify the new method of toxoplasma, rubella virus and herpes simplex virus simultaneously |
| CN106093427B (en) * | 2016-06-02 | 2018-04-13 | 滨州医学院 | The new method of Infection of Toxoplasma Gondii, rubella virus and herpes simplex virus is identified at the same time |
| WO2019169792A1 (en) * | 2018-03-07 | 2019-09-12 | 深圳市伯劳特生物制品有限公司 | Anti-torch-igm antibody repertoire chip, preparation method therefor and torch detection kit |
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