CN104360067A - Kit for quantitative detection of specific antibodies of treponema pallidum and preparation method thereof - Google Patents
Kit for quantitative detection of specific antibodies of treponema pallidum and preparation method thereof Download PDFInfo
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- CN104360067A CN104360067A CN201410629296.3A CN201410629296A CN104360067A CN 104360067 A CN104360067 A CN 104360067A CN 201410629296 A CN201410629296 A CN 201410629296A CN 104360067 A CN104360067 A CN 104360067A
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- bottle
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- treponema pallidum
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/571—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses for venereal disease, e.g. syphilis, gonorrhoea
Abstract
The invention discloses a kit for quantitative detection of specific antibodies of treponema pallidum and a preparation method thereof and relates to the treponema pallidum. The kit is provided with an external packaging box, an alkaline-phosphatase-labeled monoclonal antibody bottle, a luminous substrate bottle, 6 types of calibrator bottles containing the specific antibodies of the treponema pallidum with different concentrations, a washing liquid bottle and a recombinant-antigen-coated microporous plate, wherein the alkaline-phosphatase-labeled monoclonal antibody bottle, the luminous substrate bottle, the 6 types of calibrator bottles containing the specific antibodies of the treponema pallidum with different concentrations, the washing liquid bottle and the recombinant-antigen-coated microporous plate are arranged in the external packaging box. The preparation method comprises the following steps: preparing specific recombinant antigens of the treponema pallidum, the recombinant-antigen-coated microporous plate, natural membrane protein monoclonal antibodies, alkaline phosphatase labels of the monoclonal antibodies, a luminous substrate, a washing liquid and calibrators successively, and finally assembling into the kit for quantitative detection of the specific antibodies of the treponema pallidum. The kit can be used for quantitative detection of the specificity of the syphilis in clinical specimens and can be used as an objective index of the treating effect and the prognosis of the syphilis.
Description
Technical field
The present invention relates to microspironema pallidum, especially relate to Treponema pallidum specific antibody immue quantitative detection reagent box and preparation method thereof.
Background technology
Syphilis is the sexually transmitted disease caused by microspironema pallidum, and the incidence of disease at home rises year by year in recent years, and the prevention and control of syphilis have been classified as one of main task of China's public health service.
Microspironema pallidum can not in vitro culture, and directly aetology dark-field microscopy positive rate is not high, and Serologic test comprises cardiolipin antibody and detection of specific antibody, and the false positive rate of cardiolipin antibody is high, and sensitivity is low.And syphilis specific antibody detection method comprises fluorescent treponemal antibody absorbed test (FTA-ABS), Western blot (Western-blot) etc., its specificity is all higher, but can not quantitatively detect, the Fluctuation of patient's internal antibody cannot be judged, the result for the treatment of of disease and the objective indicator of prognosis can not be provided.
Chinese patent CN101881772A discloses a kind of microspironema pallidum based on flow microsphere carrier technique (TP) antibody test kit and preparation and determination methods method thereof, specifically comprise with TP recombinant antigen bag by high dimeric molecule microballoon, close blank binding site with bovine serum albumin(BSA), make specificity T P probe-Gao dimeric molecule microballoon; Catch TP antibody with sample Dual culture to be measured, wash the unconjugated TP antibody of centrifugal removing, then add fluorescently-labeled anti-human igg or IgM antibody; Use the fluorescence intensity of flow cytomery microballoon, qualitative or quantitative test is carried out to tested antibodies.
Summary of the invention
The object of this invention is to provide that to realize detecting microspironema pallidum quantitatively specific, for syphilis specific antibody test provides Treponema pallidum specific antibody immue quantitative detection reagent box quantitatively detected and preparation method thereof.Specifically adopting double antibody unexpectedly to strive immunoassay formats, to carry out Treponema pallidum specific antibody quantitative.
Described Treponema pallidum specific antibody immue quantitative detection reagent box is provided with external packing box, alkali phosphatase enzyme mark monoclonal antibody bottle, luminous substrate bottle, Treponema pallidum specific antibody 1000mIU/mL calibration object bottle, 500mIU/mL calibration object bottle, 200mIU/mL calibration object bottle, 50mIU/mL calibration object bottle, 10mIU/mL calibration object bottle, 0mIU/mL calibration object bottle, Washing liquid bottle, recombinant antigen bag by microwell plate, alkali phosphatase enzyme mark monoclonal antibody bottle, luminous substrate bottle, Treponema pallidum specific antibody 1000mIU/mL calibration object bottle, 500mIU/mL calibration object bottle, 200mIU/mL calibration object bottle, 50mIU/mL calibration object bottle, 10mIU/mL calibration object bottle, 0mIU/mL calibration object bottle, Washing liquid bottle, recombinant antigen bag are located in external packing box by microwell plate, alkali phosphatase enzyme mark monoclonal antibody bottle is built with alkali phosphatase enzyme mark monoclonal antibody, luminous substrate bottle is built with luminous substrate, Treponema pallidum specific antibody 1000mIU/mL calibration object bottle is built with Treponema pallidum specific antibody 1000mIU/mL calibration object, 500mIU/mL calibration object bottle is built with 500mIU/mL calibration object, 200mIU/mL calibration object bottle is built with 200mIU/mL calibration object, 50mIU/mL calibration object bottle is built with 50mIU/mL calibration object, 10mIU/mL calibration object bottle is built with 10mIU/mL calibration object, 0mIU/mL calibration object bottle is built with 0mIU/mL calibration object, Washing liquid bottle is built with Washing liquid bottle.
The preparation method of described Treponema pallidum specific antibody immue quantitative detection reagent box, comprises the following steps:
1) microspironema pallidum specificity recombinant antigen is prepared:
Adopt gene clone technology, the DNA of pcr amplification coding treponemal antigen, and insert in Escherichia coli and make it express, obtain microspironema pallidum specific antigen TPN17 and TPN47.
2) recombinant antigen bag is prepared by microwell plate
By step 1) in treponemal recombinant antigen TPN17 and TPN47 bag be buffered liquid and be diluted to 20 μ g/mL, add in microwell plate with every hole 0.1mL, 4 DEG C of bags are by 24h; Take out antigen plate, air-dry; With 1.0% skimmed milk power of 0.01mmol/L pH7.4 phosphate buffered saline, every hole 0.2mL, 4 DEG C of closed 24h, taking-up phosphate buffer washs 5 times, and room temperature is air-dry, sterilization, prepares recombinant antigen bag by microwell plate, seals for subsequent use.
3) natural membranes protein monoclonal antibody is prepared
Extract the memebrane protein of microspironema pallidum type strain Nichols strain, immune Babl/c mouse, utilizes hybridoma cell technology, the monoclonal antibody of Screening Syphilis conveyor screw specific proteins TPN17 and TPN47.
In step 3) in, described microspironema pallidum type strain Nichols strain is Treponema pallidum ssp.pallidum strain Nichol.
4) alkali phosphatase enzyme mark of monoclonal antibody
Alkaline phosphatase activates through sodium periodate, respectively with step 3)-NH2 the coupling of monoclonal antibody molecule, form alkali phosphatase enzyme mark monoclonal antibody.
5) luminous substrate preparation
Prof. Du Yucang diamantane amine luminous substrate (AMPPD) and reinforcing agent thereof, all through aseptic filtration, preparation luminous substrate working fluid.
6) cleansing solution preparation
Cleansing solution is the phosphate buffer being dissolved with Tween-20, and wherein the final concentration of Tween-20 is 0.01%.
7) calibration object
By high level Treponema pallidum specific antibody calibration object diluted, repeat demarcation three times with national standard (purchased from Products in China calibrating institute).Three calibration values, get average as the Treponema pallidum specific antibody concentration of demarcating.Described calibration object dilution is made up of 3% bovine serum albumin(BSA) of the sodium hydrogen phosphate-sodium dihydrogen phosphate of 0.05mmol/L, the sodium chloride of 0.01mmol/L, pH value 7.4.
Treponema pallidum specific antibody standard concentration series 1000mIU/mL, 500mIU/mL, 200mIU/mL, 50mIU/mL, 10mIU/mL, 0mIU/mL, be distributed into 0.5mL/ bottle, 2 ~ 8 DEG C (12 months) or-20 DEG C (4 years) deposit for subsequent use.
8) Treponema pallidum specific antibody immue quantitative detection reagent box is prepared
Alkali phosphatase enzyme mark monoclonal antibody, luminous substrate, cleansing solution, calibration object are loaded in bottle, again alkali phosphatase enzyme mark monoclonal antibody bottle, luminous substrate bottle, Washing liquid bottle, calibration object bottle and recombinant antigen bag are loaded in external packing box by microwell plate, composition Treponema pallidum specific antibody immue quantitative detection reagent box, described calibration object comprises 1000mIU/mL, 500mIU/mL, 200mIU/mL, 50mIU/mL, 10mIU/mL, 0mIU/mL totally 6 concentration.
In step 8) in, described bottle can adopt polyethylene bottle.
The invention provides a kind of Treponema pallidum specific antibody immue quantitative detection reagent box, can be used for the quantitative detection of syphilis specific in clinical samples, can be used as the objective indicator of syphilis result for the treatment of and prognosis, simple to operate, result is accurate, environment friendly and pollution-free, has long-range benefit.
The present invention adopts microspironema pallidum recombinant antigen TPN15, TPN17 as controller used in syphilis diagnosis antigen, realizes microspironema pallidum specific quantification and detects, can be used as the objective indicator of syphilis result for the treatment of and prognosis.Product of the present invention not only can carry out quantitatively the helicoid antibody in syphilitic's body, and simple to operate, and result is accurate, environment friendly and pollution-free, has long-range benefit.
Accompanying drawing explanation
Fig. 1 is the structure composition schematic diagram of Treponema pallidum specific antibody immue quantitative detection reagent box of the present invention.
Fig. 2 is standard items testing result schematic diagram.
Embodiment
Following examples will the present invention is further illustrated by reference to the accompanying drawings.
Embodiment 1
See Fig. 1, described Treponema pallidum specific antibody immue quantitative detection reagent box embodiment is provided with external packing box 1, alkali phosphatase enzyme mark monoclonal antibody bottle 2, luminous substrate bottle 3, Treponema pallidum specific antibody 1000mIU/mL calibration object bottle 4,500mIU/mL calibration object bottle 5,200mIU/mL calibration object bottle 6,50mIU/mL calibration object bottle 7,10mIU/mL calibration object bottle 8,0mIU/mL calibration object bottle 9, Washing liquid bottle 10, recombinant antigen bag by microwell plate 11, alkali phosphatase enzyme mark monoclonal antibody bottle 2, luminous substrate bottle 3, Treponema pallidum specific antibody 1000mIU/mL calibration object bottle 4,500mIU/mL calibration object bottle 5,200mIU/mL calibration object bottle 6,50mIU/mL calibration object bottle 7,10mIU/mL calibration object bottle 8,0mIU/mL calibration object bottle 9, Washing liquid bottle 10, recombinant antigen bag are located in external packing box 1 by microwell plate 11, alkali phosphatase enzyme mark monoclonal antibody bottle 2 is built with alkali phosphatase enzyme mark monoclonal antibody, luminous substrate bottle 3 is built with luminous substrate, Treponema pallidum specific antibody 1000mIU/mL calibration object bottle 4 is built with Treponema pallidum specific antibody 1000mIU/mL calibration object, 500mIU/mL calibration object bottle 5 is built with 500mIU/mL calibration object, 200mIU/mL calibration object bottle 6 is built with 200mIU/mL calibration object, 50mIU/mL calibration object bottle 7 is built with 50mIU/mL calibration object, 10mIU/mL calibration object bottle 8 is built with 10mIU/mL calibration object, 0mIU/mL calibration object bottle 9 is built with 0mIU/mL calibration object, Washing liquid bottle 10 is built with Washing liquid bottle.
Below provide the preparation method of described Treponema pallidum specific antibody immue quantitative detection reagent box, comprise the following steps:
(1) microspironema pallidum specificity recombinant antigen is prepared:
Adopt gene clone technology, the DNA of pcr amplification coding treponemal antigen, and insert in Escherichia coli and make it express, obtain microspironema pallidum specific antigen TPN17 and TPN47.
(2) recombinant antigen bag is prepared by microwell plate
Treponemal recombinant antigen TPN17 in step (1) and TPN47 bag are buffered liquid and are diluted to 20 μ g/mL, add in microwell plate with every hole 0.1mL, 4 DEG C of bags are by 24h; Take out antigen plate, air-dry; With 1.0% skimmed milk power of 0.01mmol/L pH7.4 phosphate buffered saline, every hole 0.2mL, 4 DEG C of closed 24h, taking-up phosphate buffer washs 5 times, and room temperature is air-dry, sterilization, prepares recombinant antigen bag by microwell plate, seals for subsequent use.
(3) natural membranes protein monoclonal antibody is prepared
Extract the memebrane protein of microspironema pallidum type strain Nichols strain, immune Babl/c mouse, utilizes hybridoma cell technology, the monoclonal antibody of Screening Syphilis conveyor screw specific proteins TPN17 and TPN47.
Step (3) described microspironema pallidum type strain Nichols strain is Treponema pallidum ssp.pallidum strain Nichol.
(4) alkali phosphatase enzyme mark of monoclonal antibody
Alkaline phosphatase activates through sodium periodate, respectively with the-NH2 coupling of step (3) monoclonal antibody molecule, forms alkali phosphatase enzyme mark monoclonal antibody 3.
(5) luminous substrate preparation
Prof. Du Yucang diamantane amine luminous substrate (AMPPD) and reinforcing agent thereof, all through aseptic filtration, preparation luminous substrate working fluid.
(6) cleansing solution preparation
Cleansing solution is the phosphate buffer being dissolved with Tween-20, and wherein the final concentration of Tween-20 is 0.01%.
(7) calibration object
By high level Treponema pallidum specific antibody calibration object diluted, repeat demarcation three times with national standard (purchased from Products in China calibrating institute).Three calibration values, get average as the Treponema pallidum specific antibody concentration of demarcating.Described calibration object dilution is made up of 3% bovine serum albumin(BSA) of the sodium hydrogen phosphate-sodium dihydrogen phosphate of 0.05mmol/L, the sodium chloride of 0.01mmol/L, pH value 7.4.
Treponema pallidum specific antibody standard concentration series 1000mIU/mL, 500mIU/mL, 200mIU/mL, 50mIU/mL, 10mIU/mL, 0mIU/mL, be distributed into 0.5mL/ bottle, 2 ~ 8 DEG C (12 months) or-20 DEG C (4 years) deposit for subsequent use.
(8) syphilis helicoid antibody high flux detection kit is prepared
The Treponema pallidum specific antibody immue quantitative detection reagent box that recombinant antigen bag is formed jointly by microwell plate, alkali phosphatase enzyme mark monoclonal antibody, luminous substrate, cleansing solution, calibration object and external packing box.
Described calibration object comprises 1000mIU/mL, 500mIU/mL, 200mIU/mL, 50mIU/mL, 10mIU/mL, 0mIU/mL totally 6 concentration.
Step (8) described alkali phosphatase enzyme mark monoclonal antibody, luminous substrate, cleansing solution, calibration object are all contained in corresponding polyethylene bottle.
Embodiment 2
Below provide the Treponema pallidum specific antibody in the clinical samples adopting Treponema pallidum specific antibody immue quantitative detection reagent box detection patient:
1, sample disposal: serum: venous blood 5mL, puts 37 DEG C of water-bath 30min, the centrifugal 10min of 3000g, and supernatant is for subsequent use for detecting sample.
2, application of sample: respectively add the sample of 100 μ L and standard items 1 ~ 6 in reaction plate, make blank simultaneously;
3, enzyme-added labeled monoclonal antibody: alkali phosphatase enzyme mark monoclonal antibody 100 μ L, hatches 30min for 37 DEG C;
4, wash: after 37 DEG C of reaction 30min, the syphilis specific recombinant antigen determined syphilis specific antibody and alkali phosphatase enzyme mark monoclonal antibody and microwell plate wrapping quilt strives combination, the unconjugated free composition of washing separation unexpectedly;
5, add luminous substrate working fluid, alkaline phosphatase substrate for enzymatic activity dephosphorylation base, and send the visible ray of 463nm.The relative luminous intensity unit (relative light units, RLU) respectively adding sample well is measured in 10min.The RLU of sample and syphilis specific antibody concentration negative correlation to be measured.Testing concentration in sample carries out quantitatively according to the math equation set up by determinand standard concentration and corresponding RLU, Figure 2 shows that standard items detection curve, wherein horizontal ordinate is standard concentration value, ordinate is RLU value, by sample also corresponding RLU value, the syphilis specific antibody concentration of sample to be tested can be calculated.
Claims (4)
1. Treponema pallidum specific antibody immue quantitative detection reagent box, is characterized in that being provided with external packing box, alkali phosphatase enzyme mark monoclonal antibody bottle, luminous substrate bottle, Treponema pallidum specific antibody 1000mIU/mL calibration object bottle, 500mIU/mL calibration object bottle, 200mIU/mL calibration object bottle, 50mIU/mL calibration object bottle, 10mIU/mL calibration object bottle, 0mIU/mL calibration object bottle, Washing liquid bottle, recombinant antigen bag by microwell plate, alkali phosphatase enzyme mark monoclonal antibody bottle, luminous substrate bottle, Treponema pallidum specific antibody 1000mIU/mL calibration object bottle, 500mIU/mL calibration object bottle, 200mIU/mL calibration object bottle, 50mIU/mL calibration object bottle, 10mIU/mL calibration object bottle, 0mIU/mL calibration object bottle, Washing liquid bottle, recombinant antigen bag are located in external packing box by microwell plate, alkali phosphatase enzyme mark monoclonal antibody bottle is built with alkali phosphatase enzyme mark monoclonal antibody, luminous substrate bottle is built with luminous substrate, Treponema pallidum specific antibody 1000mIU/mL calibration object bottle is built with Treponema pallidum specific antibody 1000mIU/mL calibration object, 500mIU/mL calibration object bottle is built with 500mIU/mL calibration object, 200mIU/mL calibration object bottle is built with 200mIU/mL calibration object, 50mIU/mL calibration object bottle is built with 50mIU/mL calibration object, 10mIU/mL calibration object bottle is built with 10mIU/mL calibration object, 0mIU/mL calibration object bottle is built with 0mIU/mL calibration object, Washing liquid bottle is built with Washing liquid bottle.
2. the preparation method of Treponema pallidum specific antibody immue quantitative detection reagent box as claimed in claim 1, is characterized in that comprising the following steps:
1) microspironema pallidum specificity recombinant antigen is prepared:
Adopt gene clone technology, the DNA of pcr amplification coding treponemal antigen, and insert in Escherichia coli and make it express, obtain microspironema pallidum specific antigen TPN17 and TPN47;
2) recombinant antigen bag is prepared by microwell plate
By step 1) in treponemal recombinant antigen TPN17 and TPN47 bag be buffered liquid and be diluted to 20 μ g/mL, add in microwell plate with every hole 0.1mL, 4 DEG C of bags are by 24h; Take out antigen plate, air-dry; With 1.0% skimmed milk power of 0.01mmol/L pH7.4 phosphate buffered saline, every hole 0.2mL, 4 DEG C of closed 24h, taking-up phosphate buffer washs 5 times, and room temperature is air-dry, sterilization, prepares recombinant antigen bag by microwell plate, seals for subsequent use;
3) natural membranes protein monoclonal antibody is prepared
Extract the memebrane protein of microspironema pallidum type strain Nichols strain, immune Babl/c mouse, utilizes hybridoma cell technology, the monoclonal antibody of Screening Syphilis conveyor screw specific proteins TPN17 and TPN47;
4) alkali phosphatase enzyme mark of monoclonal antibody
Alkaline phosphatase activates through sodium periodate, respectively with step 3)-NH2 the coupling of monoclonal antibody molecule, form alkali phosphatase enzyme mark monoclonal antibody;
5) luminous substrate preparation
Prof. Du Yucang diamantane amine luminous substrate and reinforcing agent thereof, all through aseptic filtration, preparation luminous substrate working fluid;
6) cleansing solution preparation
Cleansing solution is the phosphate buffer being dissolved with Tween-20, and wherein the final concentration of Tween-20 is 0.01%;
7) calibration object
By high level Treponema pallidum specific antibody calibration object diluted, repeat demarcation three times with national standard, three calibration values, get average as the Treponema pallidum specific antibody concentration of demarcating; Described calibration object dilution is made up of 3% bovine serum albumin(BSA) of the sodium hydrogen phosphate-sodium dihydrogen phosphate of 0.05mmol/L, the sodium chloride of 0.01mmol/L, pH value 7.4;
Treponema pallidum specific antibody standard concentration series 1000mIU/mL, 500mIU/mL, 200mIU/mL, 50mIU/mL, 10mIU/mL, 0mIU/mL, be distributed into 0.5mL/ bottle, deposit for subsequent use for 2 ~ 8 DEG C or-20 DEG C;
8) Treponema pallidum specific antibody immue quantitative detection reagent box is prepared
Alkali phosphatase enzyme mark monoclonal antibody, luminous substrate, cleansing solution, calibration object are loaded in bottle, again alkali phosphatase enzyme mark monoclonal antibody bottle, luminous substrate bottle, Washing liquid bottle, calibration object bottle and recombinant antigen bag are loaded in external packing box by microwell plate, composition Treponema pallidum specific antibody immue quantitative detection reagent box, described calibration object comprises 1000mIU/mL, 500mIU/mL, 200mIU/mL, 50mIU/mL, 10mIU/mL, 0mIU/mL totally 6 concentration.
3. the preparation method of Treponema pallidum specific antibody immue quantitative detection reagent box as claimed in claim 2, it is characterized in that in step 3) in, described microspironema pallidum type strain Nichols strain is Treponema pallidum ssp.pallidum strain Nichol.
4. the preparation method of Treponema pallidum specific antibody immue quantitative detection reagent box as claimed in claim 2, is characterized in that in step 8) in, described bottle adopts polyethylene bottle.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| JPS5151515A (en) * | 1974-11-01 | 1976-05-07 | Nippon Toketsu Kanso Kenkyusho | JUKAKUSETSUKETSUKYUO TANTAITOSURU BAIDOKUSETSUKETSUKYUGYOSHUHANNOJINSOKUHOYOKOGENNO SEIZOHO |
| CA2014067A1 (en) * | 1989-04-07 | 1990-10-07 | Helmut Peters | A method for the determination of antibodies against organisms causing infectious diseases in body fluids, agents for this purpose and the use thereof in this method |
| CN101363860A (en) * | 2007-08-06 | 2009-02-11 | 北京科美东雅生物技术有限公司 | Syphilis helicoid antibody chemiluminescence immune assay determination kit and method for preparing same |
| CN101738473A (en) * | 2008-11-13 | 2010-06-16 | 威海威高生物科技有限公司 | Treponema pallidum antibody diagnostic kit and preparation method thereof |
| CN102869991A (en) * | 2010-03-31 | 2013-01-09 | 积水医疗株式会社 | Reagent for assaying anti-treponema pallidum antibody |
-
2014
- 2014-11-10 CN CN201410629296.3A patent/CN104360067B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5151515A (en) * | 1974-11-01 | 1976-05-07 | Nippon Toketsu Kanso Kenkyusho | JUKAKUSETSUKETSUKYUO TANTAITOSURU BAIDOKUSETSUKETSUKYUGYOSHUHANNOJINSOKUHOYOKOGENNO SEIZOHO |
| CA2014067A1 (en) * | 1989-04-07 | 1990-10-07 | Helmut Peters | A method for the determination of antibodies against organisms causing infectious diseases in body fluids, agents for this purpose and the use thereof in this method |
| CN101363860A (en) * | 2007-08-06 | 2009-02-11 | 北京科美东雅生物技术有限公司 | Syphilis helicoid antibody chemiluminescence immune assay determination kit and method for preparing same |
| CN101738473A (en) * | 2008-11-13 | 2010-06-16 | 威海威高生物科技有限公司 | Treponema pallidum antibody diagnostic kit and preparation method thereof |
| CN102869991A (en) * | 2010-03-31 | 2013-01-09 | 积水医疗株式会社 | Reagent for assaying anti-treponema pallidum antibody |
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