CN103018441A - Multicomponent immunoassay method based on time-resolved chemiluminescence - Google Patents
Multicomponent immunoassay method based on time-resolved chemiluminescence Download PDFInfo
- Publication number
- CN103018441A CN103018441A CN2012105891569A CN201210589156A CN103018441A CN 103018441 A CN103018441 A CN 103018441A CN 2012105891569 A CN2012105891569 A CN 2012105891569A CN 201210589156 A CN201210589156 A CN 201210589156A CN 103018441 A CN103018441 A CN 103018441A
- Authority
- CN
- China
- Prior art keywords
- chemiluminescence
- antibody
- different
- time
- polynary
- 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
Images
Abstract
The invention discloses a multicomponent immunoassay method based on time-resolved chemiluminescence. The method comprises the following steps of: respectively labeling different antigens or antibodies by using chemiluminescent labels with different chemiluminescence reaction kinetic natures, and then simultaneously adding the labeled different antigens or antibodies and substances to be tested to ELISA (Enzyme-Linked Immunosorbent Assay) plate reaction holes coated with corresponding antigens or antibodies, thereby carrying out immunoreactions; and adding a co-reagent, which is used for initiating chemiluminescence, into the reaction holes, and finally, respectively detecting different chemiluminescence signals at different time windows. According to the multicomponent immunoassay method based on the time-resolved chemiluminescence, the immunoassay on different components is realized in a manner that chemiluminescence signals of the different components are respectively detected at the different time windows through time resolution, a precise fluorescence lifetime measurement system is not required for being used, and the detection on multiple components can be completed by only adopting a conventional chemiluminescence analyzer and an ordinary timing tool.
Description
Technical field
The present invention relates to the chemiluminescence immune assay field, particularly the polynary immune analysis method of time-based resolution chemical luminescent.
Background technology
Chemiluminescence immunoassay (CLIA) is the method that chemiluminescence analysis and immune response is combined foundation, and it had both had the high sensitivity of chemiluminescence analysis, had again the high specific of immunoassay.As far back as 1977, Halman etc. just created CLIA, and for detection of antigen or antibody.Because that CLIA has is highly sensitive, sensing range is wide, easy and simple to handle, detect rapidly, and realize that easily robotization detects, substitute radioactive isotope, avoided the advantages such as radioactive contamination, obtained increasingly extensive application in fields such as clinical diagnosis, environment measuring, food securities.
Along with the high speed development of immuno analytical method, often can not satisfy the actual needs that complex biological sample is detected to the detection of one-component, the appearance of multi-component immunity analytical has caused people's extensive concern.At present, existing multi-component immunity analytical method mainly can be classified as two large classes: array pattern and multi-tracer pattern.Array pattern normally detects when the different array regions of immune reactor are realized a plurality of components, as adopting array chip or sensor array.The method needs complicated instrument to do support, and usually needs to adopt expensive array-type detector, such as CCD and multi-channel electrochemical workstation.The multi-tracer pattern is another kind of common multi-component immunity analytical pattern, its ultimate principle is with different signal probe mark different components, signal by the identification different probe detects different component: as identifying with wavelength in spectroscopic methodology, identify with current potential in the electrochemical process, with specific charge identification, identify with ray energy in the radioactive method in the mass spectroscopy.
The time-resolved fluorescence multi-component immunity analytical is a kind of typical technology based on the multi-tracer pattern, this technology adopts has the rare earth metal complex of different fluorescence lifetimes as probe, then the mark different component passes through the difference of fluorescence lifetime in the detection of the existing different component of time windows cause for gossip respectively.Because fluorescence lifetime is short, utilize time-resolved fluorescence to need accurate fluorescence lifetime measurement system, expensive.Exist similar phenomenon in the chemiluminescence reaction, a lot of chemiluminescence reactions have distinct dynamic characteristic: have plenty of the flash type chemiluminescence reaction, namely signal reaches rapidly peak value, afterwards rapidly decay, fluorescent lifetime is very short, only has several seconds to several seconds zero point; Have plenty of the wide variety of glow-type chemiluminescence reaction, initial luminous signal was extremely low when namely luminescence reagent mixed, but along with the time lengthening luminous signal strengthens gradually, fluorescent lifetime from a few minutes to the dozens of minutes, or several hours to more of a specified duration; That have or concussion reaction, i.e. temporal evolution, the enhancing of chemiluminescence signal generating period and decay.But, had not yet to see report this phenomenon be used for chemiluminescence immune assay, to realize the immunoassays of various ingredients.
Summary of the invention
One of purpose of the present invention is to provide the polynary immune analysis method of time-based resolution chemical luminescent, to realize by the chemiluminescent polycomponent of time resolution.
For achieving the above object, technical scheme is:
The polynary immune analysis method of time-based resolution chemical luminescent comprises the steps:
(1) with different different antigen or the antibody of chemiluminescent labels difference mark of chemiluminescence reaction kinetic property, then adds simultaneously in the ELISA Plate reacting hole that is coated with corresponding antibodies or antigen with determinand and carry out immune response;
(2) in described reacting hole, add the chemiluminescent coreagent of initiation, then detect respectively different chemiluminescence signals at the different time window.
Among the present invention, because different chemiluminescent labels has different chemiluminescence reaction kinetic properties, therefore add the light signal that presents different component behind the coreagent at different time windows.
Preferably, described step (1) is with flash type chemiluminescent labels and different antigen or the antibody of wide variety of glow-type chemiluminescent labels difference mark, then adds simultaneously in the ELISA Plate reacting hole that is coated with corresponding antibodies or antigen with determinand and carries out immune response.
Preferably, described step (1) is with horseradish peroxidase and alkaline phosphatase difference mark Clenbuterol and Ractopamine, then joins simultaneously in the ELISA Plate reacting hole that is coated with antibody of clenbuteral and Anti-ractopamine antibody with Clenbuterol and Ractopamine and carries out immune response.
Preferably, described coated concrete grammar is: with antibody of clenbuteral and Anti-ractopamine antibody be dissolved in 0.10 M pH, the 8.0 Tris-HCl damping fluids to antibody of clenbuteral concentration be 8.0 μ g/mL, the concentration of Anti-ractopamine antibody is 10.0 μ g/mL, and mixing gets coating buffer; Then coating buffer is joined the polystyrene ELISA Plate, then place coated spending the night under 4 ℃ of conditions, remove unconjugated antibody-solutions in the hole, then every hole is cleaned with washing lotion, in each hole, add the sealing damping fluid again, 37 ℃ of lower sealings 1.5 hours, ELISA Plate is cleaned 3 times with washing lotion.
Preferably, described coreagent contains luminol, to iodophenol, and H
2O
2With alkaline phosphatase substrate liquid.
Preferably, described coreagent contains 1.0 * 10
-4The luminol of M, 5.0 * 10
-6The H to iodophenol, 10mM of M
2O
2Alkaline phosphatase substrate liquid with 25 mM.
Preferred, the adding mode of described coreagent is for adding first luminol and to iodophenol, adding H again
2O
2With the potpourri of ALP substrate solution, described H
2O
2With alkaline phosphatase substrate liquid volume ratio be 1:1.
Preferably, the pH value of described coreagent is 9.0.
The fluorescent lifetime of flash type chemiluminescent labels is very short among the present invention, only has several seconds to several seconds zero point; Wide variety of glow-type claims again to continue type, fluorescent lifetime from a few minutes to the dozens of minutes, or several hours to more of a specified duration.
Beneficial effect of the present invention: the polynary immune analysis method of time-based resolution chemical luminescent disclosed by the invention, compare with conventional fluorescence method with the spectrophotometric method of traditional multi-tracer pattern, need not to adopt beam splitting system to be sent the different wave length signal to differentiate different component; Compare with time-resolved fluorescence method, the time window of chemoluminescence method is wider: the former time window is at Millisecond, and the latter's time window from second level do not wait to hour level, therefore the time resolution for luminous signal provides more easily condition, need not to adopt complicated fluorescence lifetime measurement system, such as the precision measurement system of the relevant single photon counting technology of time-based, only need with the most common timing tool resoluting signals such as stopwatch, clocks; Compare with the multi-component immunity analytical method that adopts array pattern, the instrumentation degree is low, and is easy and simple to handle, only needs conventional chemiluminescent analyzer to get final product, and need not its complex instrument equipment commonly used and expensive array-type detector, thereby cost is lower.
Description of drawings
Fig. 1 is the schematic diagram of the chemiluminescence immune analysis method of time-based resolution of the present invention.
Fig. 2 is the working curve that the present invention detects Clenbuterol and Ractopamine.
Embodiment
In order to make the purpose, technical solutions and advantages of the present invention clearer, the below is described in detail the preferred embodiments of the present invention.
Embodiment 1
The polynary immunoassay of 1 pair of time-based resolution chemical luminescent is further described in detail by reference to the accompanying drawings.In an embodiment, with Clenbuterol (CLE) and Ractopamine (RAC) as the model analysis thing, horseradish peroxidase (HRP) and alkaline phosphatase (ALP) adopt the polynary immunoassay of time-based resolution chemical luminescent to detect different " clenbuterol hydrochloride " class materials as chemiluminescent labels difference mark CLE and RAC.The present invention adopts the competition Immune pattern, and concrete immunoassay step is as follows:
At first CLE antibody and RAC antibody are dissolved in mix in 0.10 M pH, the 8.0 Tris-HCl damping fluids after as coating buffer, to the CLE antibody concentration be 8.0 μ g/mL, the concentration of RAC antibody is 10.0 μ g/mL, then coating buffer is joined the polystyrene 96 hole ELISA Plate of high-affinity, every hole adds 100 μ L coating buffers, then place coated spending the night under 4 ℃ of conditions, remove unconjugated antibody-solutions in the hole, then every hole is cleaned 3 times with the washing lotion (containing volume fraction is the Tris-HCl buffer solution of 0.05% polysorbas20) of 300 μ L, sealing damping fluid (the SuperBlock T20 that adds again 150 μ L in each hole, Thermo Fisher Scientific Inc.) 37 ℃ of lower sealings 1.5 hours, ELISA Plate is cleaned 3 times with washing lotion; Then, in each hole, add successively 80 μ L and contain the CLE of variable concentrations and the sample solution of RAC, simultaneously adding contains the CLE antigen (10 μ g/mL) of HRP mark and each 10 μ L of RAC antigen (20 μ g/mL) of ALP mark, 1.5 h are hatched in continuation under 37 ℃, make it that competitive immunization combination occur fully.Hatch and finish rear cleaning of enzyme target, treat further detection.
In the chemiluminescence detection process, trigger simultaneously two chemiluminescence reactions by injecting coreagent.Coreagent comprises luminol, to iodophenol, and H
2O
2With the ALP substrate solution.The adding mode is 1.0 * 10 for the every hole of elder generation adds respectively 40 μ L concentration
-4The luminol of M and 20 μ L concentration are 5.0 * 10
-6M to iodophenol, then will trigger the H of triggering agent 10 mM of two chemiluminescence reactions
2O
2Mix in the ratio of 1:1 with the ALP substrate solution of 25 mM, get 30 μ L and be injected into immediately triggering reaction in the micropore to be detected, collect chemiluminescence signal by photomultiplier, the operating voltage of photomultiplier is set to-800 V, detects respectively the chemiluminescence signal of CLE and RAC at two time window places of 0.6 s and 25 min.
In the testing process of the present invention, pH value the best of coreagent is 9.0.More compatible take Tris-HCl as common buffer solution system for coreagent.As reinforcing agent, its concentration is crossed luminol-H that conference affects HRP catalysis to iodophenol
2O
2Dynamic characteristic, prolonged, therefore be preferably 5.0 * 10 in the reaction time
-6M.
Under above-mentioned experimental technique and condition, having measured concentration is CLE standard solution and the RAC standard solution of 1 ng/mL, 10 ng/mL, 100 ng/mL, 200 ng/mL, 500 ng/mL, and the result as shown in Figure 2.By testing result as can be known, high good linear relationship, its related coefficient (R of being of CLE and RAC chemical glow peak in 1.0-500 ng/mL scope
2) being respectively 0.9985 and 0.9913, detectability (S/N=2) all is 0.50 ng/mL.
Utilized the luminol-H of HRP catalysis in the present embodiment
2O
2Reaction namely reaches luminescence peak at 0.6s at certain density reinforcing agent under to the effect of iodophenol, and rapidly decay presents the flash type chemiluminescence signal afterwards; And the diamantane dioxietane phosphate hydrolysis reaction of ALP catalysis is initial luminous extremely low, along with the time lengthening luminous signal strengthens, the sustainable enhancing of whole luminescence process a few hours, presents the wide variety of glow-type chemiluminescence signal.Because both luminescence kinetics feature differences are larger, this is just for providing possible approaches with time resolution method identification different component.
In the present embodiment flash type-wide variety of glow-type chemiluminescence combination research, two individual system are carried out combinations of pairs, the result shows, presents the chemiluminescent phenomenon of time resolution, and both are better compatible, and interference is less mutually.
In order further to assess the feasibility of the method for the invention, its intersection interference is studied.During detection, in detected sample, only add a kind of analyte CLE(100 ng/mL) or RAC(100 ng/mL), then detect.Simultaneously, blank sample is detected in contrast experiment.Testing result: when detection only contained the sample of CLE, because the competitive immunoreaction of CLE and its antibody is compared with control experiment, the signal of window 1 obviously reduced, and the signal of window 2 is substantially constant.Same, when detection only contained the sample of RAC, because the competitive immunoreaction of RAC and its antibody is compared with control experiment, the signal of window 2 obviously reduced, and the signal of window 1 is substantially constant.Show thus, its intersection interference can be avoided effectively.
Embodiment 2
Take pig urine as sample, pig urine with 5 times of Tris-HCl damping fluid (pH7.0) dilutions, is added respectively CLE and RAC, to the concentration of CLE and RAC all be 10 ng/mL, 50 ng/mL and 100 ng/mL.ELISA Plate sealing and cleaning at coated CLE antibody and RAC antibody, then every hole adds respectively the pig urine dilution that 80 μ L contain CLE and RAC, simultaneously adding contains the CLE antigen (10 μ g/mL) of HRP mark and each 10 μ L of RAC antigen (20 μ g/mL) of ALP mark, under 37 ℃, hatch 1.5 h, make it that competitive immunization reaction occur.Hatch after the end again cleaning of enzyme target.Trigger chemiluminescence reaction under the reaction conditions of above-mentioned optimization, detect respectively the chemiluminescence signal of CLE and RAC at two time window places of 0.6 s and 25 min, testing result is as shown in table 1:
The recovery of standard addition of Clenbuterol and Ractopamine in the blank pig urine samples of table 1.
As shown in Table 1, the recovery of standard addition that utilizes method of the present invention to record CLE and RAC is respectively 86-104% and 88-112%, and both standard deviations all are lower than 7.6%.
Among the present invention, also can use acridinium ester-H
2O
2Detect acridinium ester-H with the combination of ALP-diamantane dioxietane phosphate system
2O
2Chemiluminescence reaction is that typical flash type is luminous, reaches peak value in the time of 0.4 second, and reaction conditions is comparatively simple, at H
2O
2Diluted alkaline solution in can be luminous.Therefore, can be combined with ALP-diamantane dioxietane phosphate system and present good chemiluminescence time resolution phenomenon.
Acridinium ester and each 10 ul of ALP are mixed in the reacting hole to be detected, and its concentration is respectively 5 * 10
-9M and 0.05 μ g/mL will contain 0.06%H
2O
2Alkaline buffer 20 ul with after ALP substrate solution 20 ul of 25 mM mix, join immediately and trigger reaction in the reacting hole to be detected, detect chemiluminescence signal by photomultiplier, located to occur first signal value in 0.4 second at first time window, along with time lengthening, the chemiluminescence intensity of second signal strengthens gradually, can select 15 min as second time window read signal.
Should be understood that, application of the present invention is not limited to above-mentioned giving an example, and for those of ordinary skills, can be improved according to the above description or conversion, and all these improvement and conversion all should belong to the protection domain of claims of the present invention.
Claims (8)
1. the polynary immune analysis method of time-based resolution chemical luminescent is characterized in that, comprises the steps:
(1) with different different antigen or the antibody of chemiluminescent labels difference mark of chemiluminescence reaction kinetic property, then adds simultaneously in the ELISA Plate reacting hole that is coated with corresponding antibodies or antigen with determinand and carry out immune response;
(2) in described reacting hole, add the chemiluminescent coreagent of initiation, then detect respectively different chemiluminescence signals at the different time window.
2. the polynary immune analysis method of described time-based resolution chemical luminescent according to claim 1, it is characterized in that: described step (1) is with flash type chemiluminescent labels and different antigen or the antibody of wide variety of glow-type chemiluminescent labels difference mark, then adds simultaneously in the ELISA Plate reacting hole that is coated with corresponding antibodies or antigen with determinand and carries out immune response.
3. the polynary immune analysis method of described time-based resolution chemical luminescent according to claim 1, it is characterized in that: described step (1) is with horseradish peroxidase and alkaline phosphatase difference mark Clenbuterol and Ractopamine, then joins simultaneously in the ELISA Plate reacting hole that is coated with antibody of clenbuteral and Anti-ractopamine antibody with Clenbuterol and Ractopamine and carries out immune response.
4. the polynary immune analysis method of described time-based resolution chemical luminescent according to claim 3, it is characterized in that: described coated concrete grammar is: with antibody of clenbuteral and Anti-ractopamine antibody be dissolved in 0.10 M pH, the 8.0 Tris-HCl damping fluids to antibody of clenbuteral concentration be 8.0 μ g/mL, the concentration of Anti-ractopamine antibody is 10.0 μ g/mL, mixing gets coating buffer; Then coating buffer is joined the polystyrene ELISA Plate, then place coated spending the night under 4 ℃ of conditions, remove unconjugated antibody-solutions in the hole, then every hole is cleaned with washing lotion, in each hole, add the sealing damping fluid again, 37 ℃ of lower sealings 1.5 hours, ELISA Plate is cleaned 3 times with washing lotion.
5. the polynary immune analysis method of each described time-based resolution chemical luminescent according to claim 1-4, it is characterized in that: described coreagent contains luminol, to iodophenol, H
2O
2With the ALP substrate solution.
6. the polynary immune analysis method of described time-based resolution chemical luminescent according to claim 5, it is characterized in that: described coreagent contains 1.0 * 10
-4The luminol of M, 5.0 * 10
-6The H to iodophenol, 10mM of M
2O
2Alkaline phosphatase substrate liquid with 25 mM.
7. the polynary immune analysis method of described time-based resolution chemical luminescent according to claim 6, it is characterized in that: the adding mode of described coreagent is for adding first luminol and to iodophenol, adding H again
2O
2With the potpourri of alkaline phosphatase substrate liquid, described H
2O
2With alkaline phosphatase substrate liquid volume ratio be 1:1.
8. the polynary immune analysis method of described time-based resolution chemical luminescent according to claim 5, it is characterized in that: the pH value of described coreagent is 9.0.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012105891569A CN103018441A (en) | 2012-12-31 | 2012-12-31 | Multicomponent immunoassay method based on time-resolved chemiluminescence |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012105891569A CN103018441A (en) | 2012-12-31 | 2012-12-31 | Multicomponent immunoassay method based on time-resolved chemiluminescence |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103018441A true CN103018441A (en) | 2013-04-03 |
Family
ID=47967280
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012105891569A Pending CN103018441A (en) | 2012-12-31 | 2012-12-31 | Multicomponent immunoassay method based on time-resolved chemiluminescence |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103018441A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106796229A (en) * | 2014-07-10 | 2017-05-31 | Fio公司 | Lateral flow/immunity-chromatography test strip service and boxlike analytical equipment, system, method and computer-readable media |
| CN107907684A (en) * | 2017-09-29 | 2018-04-13 | 何皓 | A kind of prostatic cancer early diagnosis system based on multi-fluorescence service life resolved detection technology |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1186513A (en) * | 1995-03-10 | 1998-07-01 | 梅索磅秤技术有限公司 | Multi-array, multi-specific electrochemiluminescence testing |
| US6015667A (en) * | 1996-06-03 | 2000-01-18 | The Perkin-Emer Corporation | Multicomponent analysis method including the determination of a statistical confidence interval |
| CN1908663A (en) * | 2006-08-16 | 2007-02-07 | 南京大学 | Substrate regional band resolution chemical luminescent multi-component immunity analytical method and detection system thereof |
| CN101021530A (en) * | 2007-03-26 | 2007-08-22 | 南京大学 | Automatic channel resolution chemiluminescent multicomponent immunodetection system and analytical method |
-
2012
- 2012-12-31 CN CN2012105891569A patent/CN103018441A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1186513A (en) * | 1995-03-10 | 1998-07-01 | 梅索磅秤技术有限公司 | Multi-array, multi-specific electrochemiluminescence testing |
| US6015667A (en) * | 1996-06-03 | 2000-01-18 | The Perkin-Emer Corporation | Multicomponent analysis method including the determination of a statistical confidence interval |
| CN1908663A (en) * | 2006-08-16 | 2007-02-07 | 南京大学 | Substrate regional band resolution chemical luminescent multi-component immunity analytical method and detection system thereof |
| CN101021530A (en) * | 2007-03-26 | 2007-08-22 | 南京大学 | Automatic channel resolution chemiluminescent multicomponent immunodetection system and analytical method |
Non-Patent Citations (5)
| Title |
|---|
| JOSEA.MURILLO PULGARIN: "Simultaneous stopped-flow determination of morphine and naloxone by time-resolved chemiluminescence", 《TALANTA》 * |
| JOSEA.MURILLO PULGARıN: "Simultaneous stopped-flow determination of morphine and naloxone by time-resolved chemiluminescence", 《TALANTA》, 2 October 2007 (2007-10-02) * |
| ZHENG PING LI: "Time-resolved Chemiluminescence Technique for the Microdetermination of Protein Based on their Complexation with Copper(II)", 《ANALYTICAL SCIENCES SEPTEMBER》 * |
| 张新荣: "时间分辨化学发光分析的研究I.金和银的同时化学发光测定", 《化学学报》 * |
| 武学成: "时间分辨荧光免疫分析技术及临床应用", 《医学综述》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106796229A (en) * | 2014-07-10 | 2017-05-31 | Fio公司 | Lateral flow/immunity-chromatography test strip service and boxlike analytical equipment, system, method and computer-readable media |
| CN107907684A (en) * | 2017-09-29 | 2018-04-13 | 何皓 | A kind of prostatic cancer early diagnosis system based on multi-fluorescence service life resolved detection technology |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Mathis | Rare earth cryptates and homogeneous fluoroimmunoassays with human sera | |
| CN101545007B (en) | Nano gold biological composite probe, detection method and application thereof | |
| US8956823B2 (en) | Anti-antibody reagent | |
| US11125695B2 (en) | Branched-chain amines in electrochemiluminescence detection | |
| Liu et al. | Development of a two-step immunochromatographic assay for microcystin-LR based on fluorescent microspheres | |
| Han et al. | Time-resolved chemiluminescence strategy for multiplexed immunoassay of clenbuterol and ractopamine | |
| CN103308489A (en) | Laterally-flowing immunoassay method using time resolution up-converting phosphor technology | |
| CN106706926A (en) | Serum amyloid A testing kit and manufacturing method | |
| WO1996041155A1 (en) | An improved electrochemiluminescence method | |
| CN101545864A (en) | Optical analysis reading device | |
| US9341575B2 (en) | Use of signal enhancing compounds in electrochemiluminescence detection | |
| CN102426162A (en) | Fluorescence quantitative detector | |
| JP2517102B2 (en) | Method for detecting emitted light amount of immunoassay device | |
| CN103018441A (en) | Multicomponent immunoassay method based on time-resolved chemiluminescence | |
| CN110426515A (en) | A kind of time-resolved fluoroimmunoassay chromatographic technique detects kit and its application of dirty underwater trace drugs | |
| Misiakos et al. | A multi-band capillary immunosensor | |
| CN105911041B (en) | A kind of time-resolved fluorescence detection method | |
| CN102087215A (en) | Fluorescent quantitative detection instrument | |
| Degorce | HTRF®: Pioneering technology for high-throughput screening | |
| Wang et al. | Multiplexed detection of two proteins by a reaction kinetics-resolved chemiluminescence immunoassay strategy | |
| CN1867822B (en) | Direct observation of molecular modifications in biological test systems by measuring fluorescence lifetime | |
| CN102662057A (en) | Cloned enzyme-donor immunoassay (CEDIA) ImmunoChip drug detecting kit | |
| US20160154015A1 (en) | Microfluidic device to detect cannabis in body fluids | |
| JP3922622B2 (en) | Novel fluorescent probes, methamphetamine measurement kit combining these, and simple measurement method of methamphetamine | |
| Kimura et al. | Rapid and simple quantitation of methamphetamine by using a homogeneous time-resolved fluoroimmunoassay based on fluorescence resonance energy transfer from europium to Cy5 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20130403 |