CN1431514A - Immune colloidal gold reagent for detecting IgG antibody of toxoplasma gondii and preparation method thereof - Google Patents
Immune colloidal gold reagent for detecting IgG antibody of toxoplasma gondii and preparation method thereof Download PDFInfo
- Publication number
- CN1431514A CN1431514A CN 03115145 CN03115145A CN1431514A CN 1431514 A CN1431514 A CN 1431514A CN 03115145 CN03115145 CN 03115145 CN 03115145 A CN03115145 A CN 03115145A CN 1431514 A CN1431514 A CN 1431514A
- Authority
- CN
- China
- Prior art keywords
- albumin
- preparation
- rabbit
- pad
- igg
- 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
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 20
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 title claims description 27
- 238000002360 preparation method Methods 0.000 title claims description 24
- 241000223997 Toxoplasma gondii Species 0.000 title abstract 2
- 241000283973 Oryctolagus cuniculus Species 0.000 claims abstract description 30
- 239000000020 Nitrocellulose Substances 0.000 claims abstract description 15
- 229920001220 nitrocellulos Polymers 0.000 claims abstract description 15
- 241000283707 Capra Species 0.000 claims abstract description 14
- 238000000034 method Methods 0.000 claims abstract description 13
- 238000001514 detection method Methods 0.000 claims abstract description 12
- 241000287828 Gallus gallus Species 0.000 claims abstract description 9
- 239000000084 colloidal system Substances 0.000 claims description 22
- 239000010931 gold Substances 0.000 claims description 22
- 229910052737 gold Inorganic materials 0.000 claims description 22
- 101710153593 Albumin A Proteins 0.000 claims description 19
- 239000000427 antigen Substances 0.000 claims description 9
- 102000036639 antigens Human genes 0.000 claims description 9
- 108091007433 antigens Proteins 0.000 claims description 9
- 241000223996 Toxoplasma Species 0.000 claims description 8
- 230000000274 adsorptive effect Effects 0.000 claims description 6
- 230000003053 immunization Effects 0.000 claims description 6
- 230000002829 reductive effect Effects 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 3
- 239000012141 concentrate Substances 0.000 claims description 3
- 239000002245 particle Substances 0.000 claims description 3
- 239000001509 sodium citrate Substances 0.000 claims description 3
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical class [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 claims description 3
- 235000019263 trisodium citrate Nutrition 0.000 claims description 3
- FDWREHZXQUYJFJ-UHFFFAOYSA-M gold monochloride Chemical compound [Cl-].[Au+] FDWREHZXQUYJFJ-UHFFFAOYSA-M 0.000 claims description 2
- 239000007787 solid Substances 0.000 claims description 2
- 238000010521 absorption reaction Methods 0.000 abstract description 4
- 239000012528 membrane Substances 0.000 abstract 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract 2
- 101710181478 Envelope glycoprotein GP350 Proteins 0.000 abstract 1
- 108090000623 proteins and genes Proteins 0.000 abstract 1
- 102000004169 proteins and genes Human genes 0.000 abstract 1
- 230000035945 sensitivity Effects 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 7
- 230000002950 deficient Effects 0.000 description 3
- 102000009027 Albumins Human genes 0.000 description 2
- 108010088751 Albumins Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 239000005030 aluminium foil Substances 0.000 description 2
- 238000005138 cryopreservation Methods 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 230000001900 immune effect Effects 0.000 description 2
- 238000005325 percolation Methods 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- -1 cleaning Substances 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 229940038773 trisodium citrate Drugs 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The reagent comprises a sample pad, a combination pad, a nitrocellulose membrane, a water absorption pad and a PVC backing, wherein one end of the PVC backing is sequentially adhered with the sample pad and the combination pad, the middle of the PVC backing is adhered with the nitrocellulose membrane, and the other end of the PVC backing is adhered with the water absorption pad. The conjugate pad is coated with protein A (or rabbit anti-human IgG) -colloidal gold label. The nitrocellulose membrane is coated with toxoplasma gondii specific surface membrane antigen and chicken anti-protein A (or goat anti-rabbit IgG). The reagent has the advantages of strong specificity, high sensitivity, simplicity, rapidness, low cost and capability of on-site detection, the whole process only needs 30 minutes, and operators can finish the operation according to instructions without professional training.
Description
Technical field
The present invention relates to a kind of immune colloid gold reagent that detects antibody, also relate to the preparation method of this reagent.
Background technology
The existing method that detects toxoplasma antibody IgG mainly is to detect this two kinds of equal defectiveness of method or deficiency by enzyme linked immunological absorption (EIA) and immune colloid gold percolation.The defective of enzyme linked immunological absorption (EIA) method is: need special instrument and equipment such as microplate reader to be used; Reagent needs cryopreservation; The detecting operation personnel need pass through professional training; Operating process is relatively complicated, and it is long to detect required time; It is higher to detect required expense, can not realize that single part is detected; Be unfavorable for promoting the use of in grass-roots unit.The defective of immune colloid gold percolation is: operating process is relatively complicated, can not realize single stepping; Reagent needs cryopreservation; The detection required time is long, the required expense of detection is higher.
Summary of the invention
The objective of the invention is in order to overcome the defective that current techniques exists in promoting the use of, providing a kind of does not need the auxiliary detectable of particular instrument equipment, and can reduce the detection cost effectively, alleviates the burden that needs the testing staff.The preparation method of this reagent is provided simultaneously.
Detect toxoplasma antibody IgG immune colloid gold reagent and preparation method, this reagent comprises pad, nitrocellulose filter, adsorptive pads and PVC backing, PVC backing one end adheres to sample pad, pad successively, the middle nitrocellulose filter that adheres to, and the other end adheres to adsorptive pads.Bag is by albumin A (or rabbit anti-human igg)-colloid gold label thing on the pad, and bag is by arc worm specific surfaces membranous antigen and the anti-albumin A of chicken (or goat anti-rabbit igg) on the nitrocellulose filter.If come the mark collaurum, then be used as control line with the anti-albumin A of chicken with albumin A; If come the mark collaurum with the rabbit anti-human igg, then be used as control line with goat anti-rabbit igg.
The preparation method of this reagent may further comprise the steps: (1) albumin A or rabbit anti-human igg's preparation: if use albumin A, and general just directly purchase; Rabbit anti-human igg's preparation can be by the following method: extract human antiserum's immunizing rabbit, behind the purifying the rabbit anti-human igg; (2) preparation polyclonal antibody: with arc worm specific surfaces membranous antigen immunizing rabbit repeatedly, extract the antiserum immune goat, must goat anti-rabbit igg behind the purifying; (3) preparation collaurum: the colloid gold particle that gold chloride is reduced into 20nm～40nm with reductive agents such as trisodium citrates; (4) preparation monoclonal antibody colloid gold label thing: with collaurum and albumin A (or rabbit anti-human igg) in 1: 0.005～0.015 (ml/mg) ratio mixing, make collaurum and albumin A (or rabbit anti-human igg) form stable colloidal solid, concentrate by purifying and form albumin A (or rabbit anti-human igg)-colloid gold label thing; (4) albumin A (or rabbit anti-human igg)-colloid gold label thing is coated on the collaurum pad, arc worm specific surfaces membranous antigen and the anti-albumin A of chicken (or goat anti-rabbit igg) are coated on the detection zone and the control zone of nitrocellulose filter, fully dry.
Good effect of the present invention is: cheap, production procedure is simple, and cost is low, and the expense of detection is wanted considerably cheaper than other method of use; Detection speed is fast, and overall process only needs 30 minutes, can realize that the oneself detects; Can on-the-spotly detect; Specificity is good, highly sensitive, good reproducibility; Easy and simple to handle, fast qualitative, the result is accurately, fast, and is easy and simple to handle, need not flushing process and standard control, can be in batches or single sample in time detect; Be easy to promote the use of, operating personnel need not professional training, and by specification gets final product complete operation.
Embodiment
PVC backing one end is adhered to sample pad, pad successively, the middle nitrocellulose filter that adheres to, the other end adheres to adsorptive pads.Bag is by albumin A (or rabbit anti-human igg)-colloid gold label thing on the pad.Bag is by arc worm specific surfaces membranous antigen and the anti-albumin A of chicken (or goat anti-rabbit igg) on the nitrocellulose filter, and what specifically used by mark is that albumin A or rabbit anti-human igg decide.If come the mark collaurum, then be used as control line with the anti-albumin A of chicken with albumin A; If come the mark collaurum with the rabbit anti-human igg, then be used as control line with goat anti-rabbit igg.
Preparation according to the following steps: (1) rabbit anti-human igg's preparation: extract human antiserum's immunizing rabbit, get the rabbit anti-human igg behind the purifying; (2) preparation polyclonal antibody: with arc worm specific surfaces membranous antigen immunizing rabbit repeatedly, extract the antiserum immune goat, must goat anti-rabbit igg behind the purifying; (3) preparation collaurum: the particle that 100ml 0.01% chlorauride is reduced into the 40nm size with 0.9ml 1% trisodium citrate; (4) preparation albumin A-colloid gold label thing: use 0.1mol/LK
2CO
3The pH value of colloidal gold solution is transferred to about 6.5, colloidal gold solution and monoclonal antibody are mixed in the ratio that adds the 0.8mg albumin A in the 100ml colloidal gold solution, make collaurum and antibody form stable colloidal gold composite, again by repeatedly centrifugal, abandon supernatant, cleaning, concentrate formation rabbit anti-human igg-colloid gold label thing by purifying, refrigerate standby; (5) albumin A-colloid gold label thing is sprayed on the collaurum pad, arc worm specific surfaces membranous antigen and the anti-albumin A of chicken are sprayed on the nitrocellulose filter, fully dry; (6) nitrocellulose filter, collaurum pad, sample pad, adsorptive pads etc. are bonded on the PVC backing successively; (7) the PVC material that glues is cut into the reagent strip of certain width, promptly makes and detect toxoplasma antibody IgG immune colloid gold reagent.
Before detection, earlier sample and reagent strip (plate) are placed on placement a period of time (10 minutes) under the room temperature condition, make it restore to room temperature; Take out the detectable bar from aluminium foil bag, by the direction shown in the arrow under the MARK line reagent strip is immersed in the sample solution, liquid level must not surpass the MARK line, takes out after 5 seconds～8 seconds, lies on the operator's console; If agent plate: from aluminium foil bag, take out the detectable plate, lie on the operator's console, in well, drip 3 (about 120ul) sample solutions (serum); Get final product judged result in 3 minutes～15 minutes, the result who judges after 30 minutes is invalid.The result judges: if there be " the toxoplasma antibody IgG " that will detect to exist in the sample, then red stripes appears in the detection line place, red stripes also occurs on the nature controlling line simultaneously, and this moment, the result was positive; If " the toxoplasma antibody IgG " that will not detect in the sample exists, then the detection line place does not have the band appearance, but occurs red stripes on the nature controlling line, and this moment, the result was negative.If there is not red stripes to occur on the nature controlling line, then product is invalid.
Claims (2)
1, detects toxoplasma antibody IgG immune colloid gold reagent and preparation method, this reagent comprises sample pad, pad, nitrocellulose filter, adsorptive pads and PVC backing, PVC backing one end adheres to sample pad, pad successively, the middle nitrocellulose filter that adheres to, the other end adheres to adsorptive pads, it is characterized in that bag is by albumin A (or rabbit anti-human igg)-colloid gold label thing on the pad, bag is by arc worm specific surfaces membranous antigen and the anti-albumin A of chicken (or goat anti-rabbit igg) on the nitrocellulose filter.
2, detection toxoplasma antibody IgG immune colloid gold reagent according to claim 1 and preparation method, the preparation method who it is characterized in that this reagent may further comprise the steps: (1) albumin A or rabbit anti-human igg's preparation: if use albumin A, and general just directly purchase; Rabbit anti-human igg's preparation can be by the following method: extract human antiserum's immunizing rabbit, behind the purifying the rabbit anti-human igg; (2) preparation polyclonal antibody: with arc worm specific surfaces membranous antigen immunizing rabbit repeatedly, extract the antiserum immune goat, must goat anti-rabbit igg behind the purifying; (3) preparation collaurum: the colloid gold particle that gold chloride is reduced into 20nm～40nm with reductive agents such as trisodium citrates; (4) preparation monoclonal antibody colloid gold label thing: with collaurum and albumin A (or rabbit anti-human igg) in 1: 0.005～0.015 (ml/mg) ratio mixing, make collaurum and albumin A (or rabbit anti-human igg) form stable colloidal solid, concentrate by purifying and form albumin A (or rabbit anti-human igg)-colloid gold label thing; (5) albumin A (or rabbit anti-human igg)-colloid gold label thing is coated on the collaurum pad, arc worm specific surfaces membranous antigen and the anti-albumin A of chicken (or goat anti-rabbit igg) are coated on the detection zone and the control zone of nitrocellulose filter, fully dry.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03115145 CN1431514A (en) | 2003-01-24 | 2003-01-24 | Immune colloidal gold reagent for detecting IgG antibody of toxoplasma gondii and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03115145 CN1431514A (en) | 2003-01-24 | 2003-01-24 | Immune colloidal gold reagent for detecting IgG antibody of toxoplasma gondii and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1431514A true CN1431514A (en) | 2003-07-23 |
Family
ID=4790578
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 03115145 Pending CN1431514A (en) | 2003-01-24 | 2003-01-24 | Immune colloidal gold reagent for detecting IgG antibody of toxoplasma gondii and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1431514A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103901194A (en) * | 2012-12-26 | 2014-07-02 | 深圳先进技术研究院 | Goat toxoplasmosis immunodetection test paper and preparation method thereof |
-
2003
- 2003-01-24 CN CN 03115145 patent/CN1431514A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103901194A (en) * | 2012-12-26 | 2014-07-02 | 深圳先进技术研究院 | Goat toxoplasmosis immunodetection test paper and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101173925B (en) | Colloidal gold test paper card for detecting quinolones medicament residue | |
| CN101580544B (en) | Colloidal gold labelled monoclonal antibody immunity detection Plate for detecting cannabis and tetrahydrocannabinol | |
| EP0250137A2 (en) | Colloidal gold immunoassay | |
| CN107991487A (en) | A kind of fluorescent microsphere immune test paper card, preparation and detection method for detecting carbostyril antibiotic | |
| CN201522494U (en) | Duplex occult blood fast diagnositic and chromatographic test paper | |
| CN100404553C (en) | Sturgeon family fish ovovitellin preparation method and uses | |
| CN201607444U (en) | Chromatograph test paper for fast diagnosing occult blood | |
| CN1431513A (en) | Immune colloidal gold reagent for detecting IgM antibody of toxoplasma and its preparing method | |
| CN1431509A (en) | Immune colloidal gold reagent for detecting IgM antibody of rubella virus and preparation method thereof | |
| CN1547021A (en) | Immune colloidal gold reagent for detecting chloromycetin and preparation method thereof | |
| CN1431514A (en) | Immune colloidal gold reagent for detecting IgG antibody of toxoplasma gondii and preparation method thereof | |
| CN1431510A (en) | Immune colloidal gold reagent for detecting IgG antibody of rubella virus and preparation method thereof | |
| US5310656A (en) | Vitamin B12 assay | |
| CN1431512A (en) | Immune colloidal gold reagent for detecting IgG antibody of cytomegalovirus and preparation method thereof | |
| CN1431507A (en) | Immune colloidal gold reagent for detecting IgM antibody of herpes simplex virus II type and its preparing method | |
| CN201045616Y (en) | Clip type mycoplasma pneumonia antibody detecting reagent box | |
| CN1431508A (en) | Immune colloidal gold reagent for detecting IgG antibody of herpes simplex virus II type and preparation method thereof | |
| CN107561269A (en) | The magnetic microparticle chemiluminescence detection kit and preparation method of a kind of salbutamol | |
| US5506109A (en) | Vitamin B12 assay | |
| CN1431511A (en) | Immune colloidal gold reagent for detecting IgM antibody of cytomegalovirus and its preparing method | |
| CN1431506A (en) | Immune colloidal gold reagent for detecting IgG antibody of herpes simplex virus type I and preparation method thereof | |
| CN101169413A (en) | Toxoplasma immune-gold labeled quick detection reagent and its preparation method | |
| CN1547016A (en) | Immune colloidal gold reagent for detecting sulfamethazine and preparation method thereof | |
| CN1547015A (en) | Immune colloidal gold reagent for detecting ciprofloxacin and preparation method thereof | |
| CN203178284U (en) | Triple test paper card for detecting clenbuterol hydrochloride, ractopamine and salbutamol |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| ASS | Succession or assignment of patent right |
Owner name: BODUN BIOTEST TECHNOLOGY(HANGZHOU) CO., LTD Free format text: FORMER OWNER: HUZHOU RUIZE BIOTECHNOLOGY CO., LTD Effective date: 20040325 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20040325 Address after: 311215 four road, Xiaoshan economic and Technological Development Zone, Hangzhou, Zhejiang Applicant after: Bioden Inspection Biotech (Hangzhou) Co., Ltd. Address before: 313000 Zhejiang Province, Huzhou City Longxi Road No. 208 Applicant before: Ruize Biotechnology Co., Ltd., Huzhou |
|
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |