CN103698526A - Rapid detection method and kit for mercury poisoning based on magnetic separation and quantum dot marking - Google Patents
Rapid detection method and kit for mercury poisoning based on magnetic separation and quantum dot marking Download PDFInfo
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- CN103698526A CN103698526A CN201410014482.6A CN201410014482A CN103698526A CN 103698526 A CN103698526 A CN 103698526A CN 201410014482 A CN201410014482 A CN 201410014482A CN 103698526 A CN103698526 A CN 103698526A
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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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
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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
- G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
- G01N33/542—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with steric inhibition or signal modification, e.g. fluorescent quenching
Abstract
The invention provides a rapid detection method for mercury in a liquid biological sample based on magnetic separation and quantum dots (QDs) marking. The method comprises the steps: (1) preparing mercury-resistant monoclonal antibodies; (2) coupling the mercury-resistant monoclonal antibodies with magnetic nanobeads through covalent bonds to prepare mercury-resistant immunomagnetic nanobeadd; and (3) adding a difunctional chelating agent, bovine serum albumin (BSA) and triethylamine (TEA) into the liquid biological sample to incubate, then, adding the mercury-resistant immunomagnetic nanobeads to sufficiently mix, next, carrying out magnetic separation, and adding biotinylated BSA antibodies and streptavidinylated QDs into a sediment, and carrying out fluorescence detection by using a fluorescence microplate, wherein the sediment is obtained through magnetic separation. The invention also provides a rapid detection kit for mercury in a liquid biological sample.
Description
Technical field
The invention belongs to biological and new medicine technical field.Particularly, the invention provides a kind of method and a kind of kit for fast detecting urine mercury content based on mercury content in magnetic resolution and quantum dot-labeled technology fast detecting urine.
Background technology
Mercury (Hg) is owing to having special physicochemical property, and its environmental pollution feature presents the four large distinguishing features such as persistence, animal migration, high toxicity and bioconcentration, very big to the extent of injury of human body.Yet although the toxicity of mercury is very strong, due to the restriction of the present art, mercury is still widely used in various products and technique, comprises pressure gauge, thermometer, electric switch, denture adhesive, battery and calcium carbide legal system Polyvinylchloride etc.At present, artificial mercury pollution source extensively exposes, and the mankind take in methyl mercury by diet (especially edible fishes product), by the exposure of denture adhesive and occupational environment, takes in mercury element.Meanwhile, mercury content severe overweight in some commodity (as, cosmetics), has further increased mercury poisoning victim's quantity.
At present, hospital is mainly by detecting the content of mercury in Urine in Patients, judges whether mercury poisoning of patient.The detection method of urine mercury mainly contains cold atomic absorbent spectrophotometry (AAS) and atomic fluorescence spectrometry (AFS), although these methods are ripe, credible result degree is high, but sample need to carry out complicated pre-treatment, is not suitable for the screening of fast detecting and a large amount of samples.
Immunoassay is a kind of new method that detects heavy metal ion, and detection speed is fast, highly sensitive, selectivity is strong, up to now, successfully for underwater gold, belongs to the detection of ion.Use specific bifunctional chelating agent, can chelating heavy metal ion and and carrier protein couplet, form complete immunogene, by hybridoma cell technology, prepare monoclonal antibody specific, the monoclonal antibody of preventing from heavy metal is connected to novel Fe
3o
4magnetic nanoparticle surface, can make Fe
3o
4magnetic nanoparticle has the dual-use function of target identification and magnetic resolution, thereby realizes the enrichment of heavy metal, improves detection sensitivity.
Quantum dot (QDs), is called again semiconductor nanocrystals grain, is that a kind of diameter can receive the nano particle of excitation light generation fluorescence at 1～100nm.Compare with traditional organic fluorescent dye, it is high that QDs has quantum yield, and photochemical stability is high, is difficult for the good optical characteristics such as photodissociation.Quantum dot is combined for the report of fluoroimmunoassay more and more with antibody, utilize Avidin to modify after quantum dot, quantum dot can be combined with biotinylated antibody easily, for antibody and quantum dot provide the bridge of a kind of minute sub-connection, toxin should be detected in this way, the equal detectability that carries out mark with organic dyestuff can be reached.
Summary of the invention
The present invention's application hybridoma cell technology is prepared mercury monoclonal antibody (HgMAb); The nanometer magnetic bead of activation resists and is connected with HgMAb and two respectively with QDs, prepares immune nano probe; That utilizes immunomagnetic beads can enriched character and the variation of QDs fluorescence signal, to setting up the collaborative new method of amplifying, having Hg content in supersensitive fast detecting human urine of multiple signal, has important scientific meaning and extremely wide application prospect.
Technical matters: the object of the invention is to the method for quick for heavy metal Hg in the biological sample there is no at present, set up a kind of variation that utilizes the efficient separation of immunomagnetic beads and QDs fluorescence signal, the analytical approach of Hg content in fast detecting urine.
Technical scheme: provided by the inventionly comprise the steps: based on magnetic resolution and quantum dot-labeled mercury poisoning method for quick and kit
1) Hg immunizing antigen is synthetic: cyclohexane-1 of Hg, bifunctional chelating agent [(R)-2-thiocyanogen-3-(4-aminophenyl) propyl group]-(S-S), 2 diethylenetriamine pentaacetic acids (p-SCN-Bn-CHX-A ＂-DTPA), hemocyanin (KLH) are according to a certain percentage, under triethylamine (TEA) exists, 25 ℃ of oscillating reactions 22h; Use PBS damping fluid washing reaction product, and filter unreacted small-molecule substance by super filter tube (30KD); Use above-mentioned damping fluid dilution antigen, standby.
2) Hg detectable antigens is synthetic: by Hg, p-SCN-Bn-CHX-A ＂-DTPA and bovine serum albumin(BSA) (BSA) synthetic detectable antigens under the effect of TEA.
3) preparation and purification of anti-Hg monoclonal antibody (HgMAb): prepare anti-Hg hybridoma after mouse immune, then prepare in a large number ascitic type HgMAb, finally use saturated (NH
4)
2sO
4the precipitation method are carried out antibody purification, use Sephacry S-300 to be further purified antibody.
4) anti-Hg immunomagnetic beads (HgMAb-Fe
3o
4) preparation: by HgMAb and Fe
3o
4nanometer magnetic bead coupling, prepares anti-mercury immunomagnetic beads (HgMAb-Fe
3o
4).
5) effect assessment of detection method: respectively by the range of linearity of typical curve, detection limit, precision, recovery of standard addition is tested and is used the testing result of atomic fluorescence spectrophotometer to contrast with actual sample, the accuracy of evaluation method and reliability.
Beneficial effect: mercury pollution is serious in China, set up easy, mercury poisoning method for quick not only has important theory significance and a using value to the diagnosis of toxic patient, treatment fast, also will have most important theories meaning and realistic meaning to improving China's management of public health level and public health security event disposing capacity.
The present invention is according to the double-antibody sandwich principle of fluorescence immunoassay, utilize the advantages such as immunomagnetic beads velocity of separation is fast, efficiency is high, easy and simple to handle, high in conjunction with QDs photochemical stability, be difficult for the good optical characteristics such as photodissociation, take immunomagnetic beads and QDs respectively as carrier is connected HgMAb and two anti-, the content of Hg in detection of biological sample is quickly and accurately achieved.
The present invention is only with 100 μ l urine, the reagent of tens microlitres, be less than in the time of 1h, can realize urine sample in the mensuration of mercury content.Because the instrument using is fluorescence microplate reader, can detect 96 samples simultaneously, cost is low, time-saving and efficiency.
More specifically, the invention provides the following:
1. for the kit of the fast detecting of liquid biological sample mercury, described kit comprises:
Anti-mercury immune nanometer magnetic bead, described anti-mercury immune nanometer magnetic bead will be by resisting mercury monoclonal antibody and nanometer magnetic bead to prepare via covalent bond coupling;
Bifunctional chelating agent;
Bovine serum albumin(BSA) (BSA);
Triethylamine (TEA);
Biotinylated BSA antibody; With
The QDs of streptavidin.
2. according to the kit described in 1, wherein said nanometer magnetic bead is Fe
3o
4nanometer magnetic bead.
3. according to the kit described in 1, the particle diameter of wherein said nanometer magnetic bead is 280nm.
4. according to the kit described in 1, wherein said bifunctional chelating agent is p-SCN-Bn-CHX-A ＂-DTPA.
5. according to the kit described in 1, wherein said liquid biological sample is selected from urine or blood.
6. according to the kit described in 1, wherein said liquid biological sample is urine.
7. according to the kit described in 1, wherein said anti-mercury monoclonal antibody utilizes the preparation of mercury immunizing antigen, and described mercury immunizing antigen is by mixing mercury, bifunctional chelating agent (being preferably p-SCN-Bn-CHX-A ＂-DTPA) and hemocyanin (KLH) to prepare under the existence at triethylamine (TEA).
8. according to the kit described in 1, wherein for excitation wavelength=488nm of described QDs, and for emission wavelength=585nm of described QDs.
9. the method for quick of mercury in the fluid sample based on magnetic resolution and quantum dot (QDs) mark, described method comprises:
1) prepare anti-mercury monoclonal antibody;
2) described anti-mercury monoclonal antibody and nanometer magnetic bead are passed through to covalent bond coupling, prepare anti-mercury immune nanometer magnetic bead;
3) in described liquid biological sample, add bifunctional chelating agent (being preferably p-SCN-Bn-CHX-A ＂-DTPA), bovine serum albumin(BSA) (BSA) and triethylamine (TEA), carry out incubation, add afterwards described anti-mercury immune nanometer magnetic bead fully to mix, carry out afterwards magnetic separation, the quantum dot (QDs) that adds biotinylated BSA antibody and streptavidin in the sediment obtaining to magnetic separation, is used fluorescence microplate reader to carry out fluoroscopic examination.
10. according to the method described in 9, wherein said nanometer magnetic bead is Fe
3o
4nanometer magnetic bead, its particle diameter is preferably 280nm.
Accompanying drawing explanation
Fig. 1 is mercury immunizing antigen ultraviolet spectrogram of the present invention, and Fig. 2 is mercury detectable antigens ultraviolet spectrogram of the present invention.As can be seen from the figure absorption peak is offset, and illustrates that antigen synthesizes successfully.
Fig. 3 is the anti-mercury of the mice serum of the present invention figure that tires, and Fig. 4 is the anti-mercury specificity of mice serum of the present invention figure.As can be seen from the figure, it is 32000 that the anti-mercury of mice serum is tired, and the specificity of anti-mercury is 32000, shows mouse immune success.
Fig. 5 is the anti-mercury Hybridoma Cell Culture of the present invention liquid supernatant figure that tires.We have filtered out a strain anti-mercury specificity and have tired all high cell line as seen from the figure.
Fig. 6 is the anti-mercury hybridoma of the present invention titer of ascites figure.Fig. 7 is the anti-mercury hybridoma of the present invention ascites specificity figure.As can be seen from the figure, the anti-mercury of mouse hybridoma cell ascites is tired and specificity all can reach 25600, illustrates and in ascites, has specific anti-mercury antibody.
Fig. 8 is the purified anti-mercury monoclonal antibody figure that tires.As can be seen from the figure the anti-mercury monoclonal antibody after, purified is tired higher.
Fig. 9 is anti-mercury monoclonal antibody gel electrophoresis figure after purifying.As can be seen from the figure the antibody molecule amount of extracting is 25KDa and 50KDa, there is no abnormal band, and purity is higher.
Figure 10 is the schematic diagram of nanometer magnetic bead and monoclonal antibody reactive.
Embodiment
By following examples, further illustrate the present invention, but claim of the present invention is not limited only to embodiment.
1) Hg immunizing antigen is synthetic: by Hg (national non-ferrous metal and electronic material Institute of Analysis, GSB04-1729-2004), cyclohexane-1 of bifunctional chelating agent [(R)-2-thiocyanogen-3-(4-aminophenyl) propyl group]-(S-S), 2 diethylenetriamine pentaacetic acids (p-SCN-Bn-CHX-A ＂-DTPA) (Macroyclics, the U.S.), hemocyanin (KLH) (Sigma, the U.S., H7017), triethylamine (TEA) is according to the ratio of Hg:p-SCN-Bn-CHX-A ＂-DTPA:KLH:TEA=9:12:200:13 (W/W/W/W), 25 ℃ of oscillating reactions 22h; Use PBS damping fluid washing reaction product, and filter unreacted small-molecule substance by super filter tube (30KD); The Hg immunizing antigen that uses the dilution of PBS damping fluid to obtain.
2) Hg detectable antigens is synthetic: by Hg, p-SCN-Bn-CHX-A ＂-DTPA, bovine serum albumin(BSA) (BSA) (Sigma, the U.S., A1933) synthesizes detectable antigens with TEA according to the ratio of Hg:p-SCN-Bn-CHX-A ＂-DTPA:BSA:TEA=9:12:200:13 (W/W/W/W).The evaluation of immunizing antigen and detectable antigens as shown in Figure 1, 2.
The preparation and purification of anti-Hg monoclonal antibody (HgMAb):
By the Hg immunizing antigen of 1.0mg/ml and equal-volume Freund's complete adjuvant (FCA), by the two pushing manipulations of syringe, fully mix, make it form white water in oil emulsion shape thing, i.e. antigen emulsifying agent.Use multiple spot immunization, immunity female Balb/C mouse in 6 week age (Beijing Vital River Experimental Animals Technology Co., Ltd., 211), every immunity in 2 weeks 1 time, immunity is 5 times altogether.Carrying out the 2nd, 3,4 times when immune, Hg immunizing antigen is mixed with equal-volume incomplete Freund's adjuvant (FIA), with same dosage booster immunization, during the 5th immunity, only with Hg immunizing antigen, do not add adjuvant.After mouse immune mice serum tire with specificity as shown in Figure 3-4.Get the immune mouse that antiserum titre is the highest and prepare splenocyte, according to list of references (Howard.G.C., Kaser.M.R. antibody preparation and use experiment guide [M]. Beijing: Science Press, 2010.) by splenocyte and SP2/0 murine myeloma cell (Nanjing KaiJi Biology Science Development Co., Ltd, KG075) merge, screening positive hybridoma cell is cloned, and after positive rate reaches 100%, cell is expanded and is cultivated, and result as shown in Figure 5.Positive cell is expelled to mouse peritoneal, extracts ascites, evaluate titer of ascites, result as shown in Figure 6,7.Ascites is used respectively saturated (NH
4)
2sO
4pbMAb in the precipitation method and Sephacry S-300 column chromatography purification ascites.The antibody of purifying is carried out to bioactivity and gel electrophoresis experiment, and result respectively as shown in FIG. 8 and 9.
Embodiment 3
Anti-Hg immunomagnetic beads (HgMAb-Fe
3o
4) preparation: because nanometer magnetic bead (
m-280Tosylactivated, lifetechnologies, the U.S., 14203) background value is low, and antibody and bead surface are covalently bound, are the splendid selections that makes protein complex immunoprecipitation.Microballon magnetic is assembled gentle and quick and is hatched consuming time extremely short.Therefore nanometer magnetic bead is selected
m-280Tosylactivated, its particle diameter is 280nm.Anti-Hg immunomagnetic beads synthesis step is as follows:
(1) get 165 μ L nanometer magnetic beads, the separated 1min of magnetic, removes supernatant;
(2) add the phosphate buffer reaction of the anti-Hg monoclonal antibody of 100 μ g and 0.1M pH7.4 to make volume become 150 μ L, vortex concussion;
(3) add 100 μ L3M (NH
4)
2sO
4, 0.1M pH7.4 phosphate buffer vortex concussion;
(4) 37 ℃ are shaken hatching 12-18h;
(5) the separated 2min of magnetic, removes supernatant;
(6) remove magnet, add the phosphate buffer of 1mL0.5%BSA0.01M pH7.4,37 ℃ are shaken hatching 1h;
(7) the separated 2min of magnetic, removes supernatant;
(8) remove magnet, add the phosphate buffer of 1mL0.1%BSA0.01M pH7.4, vortex vibration 5-10s.
(9) the separated 2min of magnetic, removes supernatant;
(10) repeating step 7-8;
(11) with the resuspended immunomagnetic beads of phosphate buffer of 240 μ L0.1%BSA0.01M pH7.4.
Nanometer magnetic bead and anti-Hg monoclonal antibody reactive principle are as shown in figure 10.
Embodiment 4
Method for quick based on magnetic resolution and quantum dot-labeled mercury poisoning:
(1) in 0.1mL urine sample, add 13 μ l10 μ g/ml p-SCN-Bn-CHX-A ＂-DTPA, 1 μ l2.17mg/ml BSA and 1 μ l4.6 * 10
-4%TEA, 37 ℃ of shaking table reaction 15min.
(2) add the separated 2min of magnetic after the anti-Hg immunomagnetic beads of 2 μ L reaction 15min.
(3) add the biotinylated BSA antibody of 4 μ L (abcam, Britain, the streptavidin of ab7636) He 1 μ L1mM obtain quantum dot (
585Streptavidin Conjugate, lifetechnologies, the U.S., Q10113MP), the separated 2min of magnetic after 37 ℃ of shaking table reaction 15min.
(4) with fluorescence microplate reader, carry out result detection (Ex=488nm, Em=585nm).
Standard curve determination: get the blank urine sample 100 μ L of Healthy People in 200 μ L centrifuge tubes, add the Hg of variable concentrations to be mixed with concentration and be respectively 1,10,100,500 and the standard urine sample of 1000ng/mL.By method for quick of the present invention, process, with concentration (C), absorbance (OD) is carried out to linear regression, calculate typical curve equation C=59.491OD-535.052, R=0.963.This detection method range of linearity is 1-1000ng/ml, and minimum detectability is 1ng/ml.
Recovery of standard addition is measured: after adding respectively 50ng Hg then it to be processed according to method for quick patient's 0.1mL blood sample (BEIJING CHAO-YANG HOSPITAL's occupational illness and poisoning medical center), detect, ratio by the amount of recording with addition, calculates recovery of standard addition.Recovery of standard addition is 94.70%-101.18%.
Precision is measured: the urine sample of the basic, normal, high 3 kinds of concentration of preparation Hg, according to method for quick of the present invention, to process, and sample introduction is measured.In 1 day, operation repetitive is 5 times, calculates withinday precision.Result is as shown in table 1.
Table 1: Mercury Determination precision (n=5) in urine sample
Getting urine sample (BEIJING CHAO-YANG HOSPITAL's occupational illness and poisoning medical center) detects and contrasts evaluation accuracy with atomic fluorescence method according to method for quick of the present invention, difference not statistically significant between the measurement result of two kinds of methods, show that the method accuracy of setting up is reliable, result is as shown in table 2.
Table 2: two kinds of detection methods are to mercury content testing result comparison (ng/mL) in urine sample
Detection method specificity identification: heavy metal monoclonal antibody specificity shows with antibody cross reaction: measure antibody and other metallic ions Hg obtaining according to reaction conditions
2+, Pb
2+, Mn
2+, Bi
2+, Ni
2+, whether react or react size cases.By after the continuous concentration gradient dilution of Hg, DTPA reacts 30min with sequestrant, then according to method for quick of the present invention, detects, and sets up all metals and suppresses typical curve, and calculate half and press down numeral system rate (IC
50), cross reacting rate (CR) calculates by formula: cross reacting rate (CR)=IC
50(Pb)/IC
50(other metallic ion) * 100% use half suppresses (IC
50) weigh antibody sensitivity, it is lower that half suppresses, and illustrates that the sensitivity of antibody is higher; Also be IC
50numerical value less, illustrate that the specific performance of antibody is stronger.Table 3 shows the result of anti-plumbous monoclonal antibody of the present invention and other metallic ion cross reactions, and the cross reacting rate of anti-plumbous monoclonal antibody of the present invention and other metallic ions is lower as can be seen from Table 3.
The cross reaction of the anti-mercury monoclonal antibody of table 3 and other ions
Claims (10)
1. for the kit of the fast detecting of liquid biological sample mercury, described kit comprises:
Anti-mercury immune nanometer magnetic bead, described anti-mercury immune nanometer magnetic bead will be by resisting mercury monoclonal antibody and nanometer magnetic bead to prepare via covalent bond coupling;
Bifunctional chelating agent;
Bovine serum albumin(BSA) (BSA);
Triethylamine (TEA);
Biotinylated BSA antibody; With
The QDs of streptavidin.
2. kit according to claim 1, wherein said nanometer magnetic bead is Fe
3o
4nanometer magnetic bead.
3. kit according to claim 1, the particle diameter of wherein said nanometer magnetic bead is 280nm.
4. kit according to claim 1, wherein said bifunctional chelating agent is p-SCN-Bn-CHX-A ＂-DTPA.
5. kit according to claim 1, wherein said liquid biological sample is selected from urine or blood.
6. kit according to claim 1, wherein said liquid biological sample is urine.
7. kit according to claim 1, wherein said anti-mercury monoclonal antibody utilizes the preparation of mercury immunizing antigen, and described mercury immunizing antigen is by mixing mercury, bifunctional chelating agent (being preferably p-SCN-Bn-CHX-A ＂-DTPA) and hemocyanin (KLH) to prepare under the existence at triethylamine (TEA).
8. kit according to claim 1, wherein for excitation wavelength=488nm of described QDs, and for emission wavelength=585nm of described QDs.
9. the method for quick of mercury in the fluid sample based on magnetic resolution and quantum dot (QDs) mark, described method comprises:
1) prepare anti-mercury monoclonal antibody;
2) described anti-mercury monoclonal antibody and nanometer magnetic bead are passed through to covalent bond coupling, prepare anti-mercury immune nanometer magnetic bead;
3) in described liquid biological sample, add bifunctional chelating agent (being preferably p-SCN-Bn-CHX-A ＂-DTPA), bovine serum albumin(BSA) (BSA) and triethylamine (TEA), carry out incubation, add afterwards described anti-mercury immune nanometer magnetic bead fully to mix, carry out afterwards magnetic separation, the quantum dot (QDs) that adds biotinylated BSA antibody and streptavidin in the sediment obtaining to magnetic separation, is used fluorescence microplate reader to carry out fluoroscopic examination.
10. method according to claim 9, wherein said nanometer magnetic bead is Fe
3o
4nanometer magnetic bead, its particle diameter is preferably 280nm.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104965076A (en) * | 2015-07-14 | 2015-10-07 | 上海拜豪生物科技有限公司 | Cadmium-chelated immune complex as well as preparation method and application thereof |
| CN105510287A (en) * | 2015-11-30 | 2016-04-20 | 环境保护部华南环境科学研究所 | Signal-amplified high-sensitivity detection method for Hg<2+> in water environments |
| CN106483121A (en) * | 2016-08-26 | 2017-03-08 | 赵红梅 | Extract verifying attachment in metal poisoning body |
| CN108383903A (en) * | 2018-02-09 | 2018-08-10 | 浙江工商大学 | The application of a kind of preparation method and DOTA of heavy metal Hg artificial antigen in preparing heavy metal Hg artificial antigen reagent |
| CN113341149A (en) * | 2021-06-09 | 2021-09-03 | 军事科学院军事医学研究院环境医学与作业医学研究所 | Kit and detection method for detecting fluoroquinolone antibiotics |
| CN115950877A (en) * | 2022-10-20 | 2023-04-11 | 国家粮食和物资储备局科学研究院 | Magnetic particle chemiluminescence kit for detecting heavy metals |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004323508A (en) * | 2003-03-20 | 2004-11-18 | Central Res Inst Of Electric Power Ind | Anti-heavy metal monoclonal antibody and method for using the same |
| CN101441212A (en) * | 2008-12-04 | 2009-05-27 | 上海交通大学 | Multiple-antigen synchronous detection method of quantum dot mark fluorescent immune |
| CN101486767A (en) * | 2009-03-02 | 2009-07-22 | 华南理工大学 | Mercury resistant chelate monoclonal antibody, as well as preparation method and application thereof |
| US20100167333A1 (en) * | 2006-01-12 | 2010-07-01 | Life Technologies Corporation | Heavy Metal Binding Compounds and Their Method of Use |
| CN101948807A (en) * | 2010-08-24 | 2011-01-19 | 北京市农林科学院 | Heavy metal mercury ion antibody and application thereof |
| CN103472231A (en) * | 2013-09-28 | 2013-12-25 | 郑州大学 | Indirect competition enzyme linked immunoreagent kit for detecting mercury ions and manufacturing method thereof |
| CN103499688A (en) * | 2013-09-27 | 2014-01-08 | 河南科技学院 | Gold-labeled test strip/card for quickly detecting mercury ions |
-
2014
- 2014-01-13 CN CN201410014482.6A patent/CN103698526A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004323508A (en) * | 2003-03-20 | 2004-11-18 | Central Res Inst Of Electric Power Ind | Anti-heavy metal monoclonal antibody and method for using the same |
| US20100167333A1 (en) * | 2006-01-12 | 2010-07-01 | Life Technologies Corporation | Heavy Metal Binding Compounds and Their Method of Use |
| CN101441212A (en) * | 2008-12-04 | 2009-05-27 | 上海交通大学 | Multiple-antigen synchronous detection method of quantum dot mark fluorescent immune |
| CN101486767A (en) * | 2009-03-02 | 2009-07-22 | 华南理工大学 | Mercury resistant chelate monoclonal antibody, as well as preparation method and application thereof |
| CN101948807A (en) * | 2010-08-24 | 2011-01-19 | 北京市农林科学院 | Heavy metal mercury ion antibody and application thereof |
| CN103499688A (en) * | 2013-09-27 | 2014-01-08 | 河南科技学院 | Gold-labeled test strip/card for quickly detecting mercury ions |
| CN103472231A (en) * | 2013-09-28 | 2013-12-25 | 郑州大学 | Indirect competition enzyme linked immunoreagent kit for detecting mercury ions and manufacturing method thereof |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104965076A (en) * | 2015-07-14 | 2015-10-07 | 上海拜豪生物科技有限公司 | Cadmium-chelated immune complex as well as preparation method and application thereof |
| CN105510287A (en) * | 2015-11-30 | 2016-04-20 | 环境保护部华南环境科学研究所 | Signal-amplified high-sensitivity detection method for Hg<2+> in water environments |
| CN105510287B (en) * | 2015-11-30 | 2019-01-29 | 环境保护部华南环境科学研究所 | Hg in a kind of highly sensitive water environment of signal amplification2+Detection method |
| CN106483121A (en) * | 2016-08-26 | 2017-03-08 | 赵红梅 | Extract verifying attachment in metal poisoning body |
| CN108383903A (en) * | 2018-02-09 | 2018-08-10 | 浙江工商大学 | The application of a kind of preparation method and DOTA of heavy metal Hg artificial antigen in preparing heavy metal Hg artificial antigen reagent |
| CN113341149A (en) * | 2021-06-09 | 2021-09-03 | 军事科学院军事医学研究院环境医学与作业医学研究所 | Kit and detection method for detecting fluoroquinolone antibiotics |
| CN113341149B (en) * | 2021-06-09 | 2023-08-22 | 军事科学院军事医学研究院环境医学与作业医学研究所 | Kit and detection method for detecting fluoroquinolone antibiotics |
| CN115950877A (en) * | 2022-10-20 | 2023-04-11 | 国家粮食和物资储备局科学研究院 | Magnetic particle chemiluminescence kit for detecting heavy metals |
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