CN1340712A - Reagent kit for enzyme-linked immunoassay of rubella, macrocell and monospermous viruses and Toxoplasma antibody - Google Patents
Reagent kit for enzyme-linked immunoassay of rubella, macrocell and monospermous viruses and Toxoplasma antibody Download PDFInfo
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- CN1340712A CN1340712A CN 00123730 CN00123730A CN1340712A CN 1340712 A CN1340712 A CN 1340712A CN 00123730 CN00123730 CN 00123730 CN 00123730 A CN00123730 A CN 00123730A CN 1340712 A CN1340712 A CN 1340712A
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Abstract
A reagent kit for enzyme-linked immunoassay is disclosed. In the procedure to diagnose early infection of ToRCH, a single test can fully screen out the negative specimens of ToRCH.IgM. The reagent kit of ToRCH.IgM antibody is composed of human IgM antibody coated plate, composite ToRCH antigen, combined enzyme tag, substrate liquid reaction terminating liquid, washing liquid, and negative and positive reference serum. Its advantages are simple test and low cost.
Description
The invention belongs to biological technical field.
The inspection-free survey method of enzyme is characteristic such as quick, easy and sensitivity height with it, has obtained increasingly extensive application in the diagnostic detection in worldwide, and has progressively replaced the solid phase RIA method.
Arc worm (Toxoplasma), rubella virus (Rubella virus), cytomegalovirus (Cytomegalovirus), herpes simplex virus I-type, II type (Herpes simplex virus I, II), be abbreviated as ToRCH, " national clinical examination working specification " (second edition) existing regulation below is promptly used this abbreviation.
It all is that one-time detection can only be surveyed an index that prior art detects the agent of being excused from an examination of enzyme that ToRCH infects, not only wastes reagent like this in actual detected, and efficient can not improve, and uses inconvenience, and other method also has this disadvantage.
The present invention relates in the process of clinical diagnose ToRCH early infection, pass through single experiment, can all screen out the negative sample of ToRCHIgM, a kind of enzyme that the joint-detection of ToRCHIgM antibody is achieved is exempted from diagnostic kit, mainly solve in the diagnosis ToRCH early infection process, usually need detect individual event antibody one by one to a sample, not only time-consuming but also require great effort, increase the problem of reagent cost again.
Feature of the present invention is to exempt under the working condition of diagnostic reagent at general enzyme, finish the ToRCHIgM abzyme and exempt from the assembly of joint inspection kit, adopt enzyme to exempt from prize law, kit consists of anti-human IgM antibody bag by plate, the multinomial combined antigen of ToRCH, multinomial combination enzyme labeling thing, substrate solution A (TMB), substrate solution B (H
2O
2), reaction terminating liquid (H
2SO
4), cleansing solution (PBST), the positive and negative control serum, by to two to five species-specific antigens and corresponding two to five species specificity enzyme labelled antibodies, carry out assembly by proper proportion and concentration, make all negative sample of individual event ToRCH index, the joint inspection result is still negative, and single index is then positive if any the sample joint inspection result of the positive; The negative sample that accounts for 90% ratio like this in the clinical samples can be defined as feminine gender, and the positive sample of 10% left and right sides ratio is exempted from kit with the individual event enzyme again and examined and determine one by one, to determine concrete positive index.
The present invention relates to five joint inspection kits of ToRCH, four joint inspection kits, three joint inspection kits, binomial joint inspection kit, all can once distinguish feminine gender or positive IgM antibody.
Below in conjunction with embodiment the present invention is described.
With toxoplasma gondii (Toxo), rubella virus (RV), cytomegalovirus (CMV), herpes simplex virus I-type, II type (HSV-I), (HSV-II) five IgM antibody joint inspection kit is example.
This kit adopts enzyme to exempt from prize law, and this reagent is achieved through the following technical solutions:
1, with the anti-human IgM antibody (μ chain) of optimum concentration bag by the reaction plate micropore and washed, shrouding.
2, assembly ToRCH combined antigen is determined the optimal proportion and the concentration of assembly by titration.(agent of being excused from an examination of common individual event enzyme is an individual event antigen, and we have used two to five joint inspection antigens of ToRCH in the joint inspection reagent, and embodiment is five joint inspections, and therefore combined antigen herein is Toxo, RV, CMV, HSV-I, five combined antigens of HSV-II.)
3, preparation Toxo, RV, CMV, HSV-I, five required enzyme labeling things of joint inspection of HSV-II solution, used every antibody all need reach certain purity, (HRP) carries out mark with horseradish peroxidase, by drop reaction, determines the concentration of final enzyme labeling thing solution.
4, the preparation of substrate solution, selecting tetramethyl benzidine (TMB) for use is substrate solution A, hydrogen peroxide (H
2O
2) be substrate solution B.
5, normal human serum is adopted in negative serum preparation, and many parts are mixed and carry out preservative treatment.
6, the serum of the ToRCHIgM antibody test positive is adopted in the preparation of positive serum, and many parts are mixed and carry out and adopt after deactivation and the preservative treatment.
7, cleansing solution and stop buffer are selected the desired universal method preparation of conventional ELISA method for use, and cleansing solution is PBST, and stop buffer is 2MH
2SO
4
8, the kit of this composition carries out operation detection according to the ELISA method, and 10 routine clinical samples results are listed below table:
P/N＞5 reagent are effective, and N value＜0.05 o'clock is 0.05 in N,
Sample OD value＞2.1N is judged to the positive.
Wherein, No. 3, joint inspection result, No. 9 are positive, and all the other are all negative;
Individual event kit result is:
No. 3: the CMVIgM positive
No. 9: ToxoIgM, the RVIgM positive
The present invention can be applied to clinical diagnose simply and easily, especially greatly reduces testing amount and reagent cost, for large-scale operation provides more simple and effective method.
Claims (3)
1, rubella, giant cell, herpes simplex virus and toxoplasma antibody (ToRCHIgM) enzyme is exempted from the joint inspection kit, it is characterized in that this kit is in the process of diagnosis ToRCH early infection, by single experiment, can all screen out the negative sample of ToRCHIgM, the antibody combined detection of ToRCHIgM is achieved, kit basic composition is: the positive and negative control serum, anti-human IgM antibody bag is by plate, the multinomial combined antigen of ToRCH, the multinomial combination enzyme labeling of ToRCH thing, substrate solution A (TMB), substrate solution B (H
2O
2), stop buffer (H
2SO
4), cleansing solution (PBST).
2, joint inspection kit as claimed in claim 1 is characterized in that realizing two to five any joint inspections of ToRCHIgM antibody.
3, joint inspection kit as claimed in claim 1 is characterized in that allocating suitable neutralization combined antigen and specificity group synthase label, and its ratio should make the joint inspection result detect consistent with individual event with concentration.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 00123730 CN1340712A (en) | 2000-09-01 | 2000-09-01 | Reagent kit for enzyme-linked immunoassay of rubella, macrocell and monospermous viruses and Toxoplasma antibody |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 00123730 CN1340712A (en) | 2000-09-01 | 2000-09-01 | Reagent kit for enzyme-linked immunoassay of rubella, macrocell and monospermous viruses and Toxoplasma antibody |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1340712A true CN1340712A (en) | 2002-03-20 |
Family
ID=4590087
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 00123730 Pending CN1340712A (en) | 2000-09-01 | 2000-09-01 | Reagent kit for enzyme-linked immunoassay of rubella, macrocell and monospermous viruses and Toxoplasma antibody |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1340712A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1300587C (en) * | 2004-12-20 | 2007-02-14 | 山东省医药生物技术研究中心 | Protein chip for detecting blood and cerebro spinal fluid pathogen antibody, and its preparing method and use |
| CN100554965C (en) * | 2005-08-05 | 2009-10-28 | 珠海经济特区海泰生物制药有限公司 | Toxoplasmosis IgM antigen testing reagent and application |
| CN104155449A (en) * | 2014-07-28 | 2014-11-19 | 广州市丰华生物工程有限公司 | Method and kit for detecting TORCH IgM antibodies and preparation method of kit |
| CN106153894A (en) * | 2016-08-12 | 2016-11-23 | 江苏泽成生物技术有限公司 | A kind of detection kit measuring Toxoplasma Gondi IgM antibody and detection method thereof |
| CN109001468A (en) * | 2018-08-06 | 2018-12-14 | 滴准生物科技（常州）有限公司 | A kind of detection kit of CMV antibody IgM |
| CN109116031A (en) * | 2018-07-25 | 2019-01-01 | 滴准生物科技（常州）有限公司 | A kind of detection kit can detect a variety of prenatal and postnatal care pathogen IgG antibodies |
-
2000
- 2000-09-01 CN CN 00123730 patent/CN1340712A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1300587C (en) * | 2004-12-20 | 2007-02-14 | 山东省医药生物技术研究中心 | Protein chip for detecting blood and cerebro spinal fluid pathogen antibody, and its preparing method and use |
| CN100554965C (en) * | 2005-08-05 | 2009-10-28 | 珠海经济特区海泰生物制药有限公司 | Toxoplasmosis IgM antigen testing reagent and application |
| CN104155449A (en) * | 2014-07-28 | 2014-11-19 | 广州市丰华生物工程有限公司 | Method and kit for detecting TORCH IgM antibodies and preparation method of kit |
| CN106153894A (en) * | 2016-08-12 | 2016-11-23 | 江苏泽成生物技术有限公司 | A kind of detection kit measuring Toxoplasma Gondi IgM antibody and detection method thereof |
| CN109116031A (en) * | 2018-07-25 | 2019-01-01 | 滴准生物科技（常州）有限公司 | A kind of detection kit can detect a variety of prenatal and postnatal care pathogen IgG antibodies |
| CN109001468A (en) * | 2018-08-06 | 2018-12-14 | 滴准生物科技（常州）有限公司 | A kind of detection kit of CMV antibody IgM |
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|---|---|---|---|
| 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 |