CN202956384U - Up-converting phosphor rapid ration kit used for neutrophil gelatinase associated lipocalin (NGAL) detection - Google Patents
Up-converting phosphor rapid ration kit used for neutrophil gelatinase associated lipocalin (NGAL) detection Download PDFInfo
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- CN202956384U CN202956384U CN 201220497401 CN201220497401U CN202956384U CN 202956384 U CN202956384 U CN 202956384U CN 201220497401 CN201220497401 CN 201220497401 CN 201220497401 U CN201220497401 U CN 201220497401U CN 202956384 U CN202956384 U CN 202956384U
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Abstract
The utility model discloses an up-converting phosphor rapid ration kit used for neutrophil gelatinase associated lipocalin (NGAL) detection and belongs to the technical field of clinical medical detection. The up-converting phosphor rapid ration kit used for the NGAL detection comprises a sample pad, a combination pad, an analysis membrane and a piece of absorbent paper, wherein the sample pad, the combination pad, the analysis membrane and the piece of absorbent paper are stuck on a bottom liner in sequence. NGAL antibody I marked by up-converting phosphor uncoupling protein (UCP) particles is coated on the combination pad. A detection area and a quality control area are arranged on the analysis membrane, wherein the detection area is parallel to the quality control area. NGAL antibody II or NGAL antigen is coated on the detection area. Goat anti mouse IgG is coated on the quality control area. Due to the fact that the up-converting phosphor technique and the immunological technique are combined, the up-converting phosphor rapid ration kit used for the NGAL detection has the advantages of being accurate and rapid in detection and ration detection is achieved.
Description
Technical field
The utility model relates to the clinical medical inspection field, is specifically related to a kind ofly fast, quantitatively detect the NGAL kit based on upper forwarding light technology immunochromatography.
Background technology
Upper forwarding light particle (Up-Converting Phosphor, UCP) is the rare earth metal complex that turns on a kind of can the carrying out energy, and namely UCP can absorb low-energy (long wavelength) infrared light, but sends high-octane (short wavelength) visible light.UCP is mixed in the lattice of some crystal by several thuliums to consist of.Three kinds of main compositions are arranged: main matrix, absorption and emission in this material.Crystalline material as main matrix has: oxysulfide is (such as Y
2O
2S, GdO
2S, La
2O
2S etc.), fluoride is (such as YF
3, GdF
3, LaF
3Deng), gallate is (such as YGaO
3, Y
3Ga
5O
12Deng) and silicate (YSi
2O
5, YSi
3O
7Deng) etc.; Being commonly used for the rare earth ion that absorbs son has: ytterbium ion (Yb
3+), erbium ion (Er
3+), samarium ion (Sm
3+) etc.; The rare earth ion that is commonly used for emission has: erbium ion (Er
3+), holmium ion (Ho
3+), thulium ion (Tm
3+), terbium ion (Tb
3+) etc.Spatial orientation and distance that absorption and this ion pair of emission suit in the main matrix lattice are to produce upper basis of transmitting light.
This unique optical properties that upper forwarding light particle UCP has makes it bring into play huge potential in bedside quick diagnosis field:
⑴ the sensitivity of height: exclusive upper forwarding optical phenomenon has guaranteed that there is not the background interference that comes from the outside in UCP in testing process;
⑵ the stability of height: the luminescence phenomenon of UCP is the pure physical process that results from inside configuration, thereby has avoided the luminous cancellation from test sample is corroded and self decays and causes fully;
⑶ high degree of flexibility: but the diversified characteristic spectrum of the independent assortment that UCP has makes it be applicable to multiple analysis;
⑷ the security of height: inorganic inertia, infrared ray excited, VISIBLE LIGHT EMISSION are so that detect for tester, detected sample, environment all without endangering based on UCP.
Neutrophil gelatinase-associated lipocalin (neutrophil gelatinase-associated lipocalin, NGAL) be 1993 found first in neutrophil leucocyte, with inflammation, embryonic development, immune response, chemotaxis, signal transduction and kinds of tumors to betide the processes such as development relevant.
Recent domestic studies show that NGAL albumen has the characteristics of specific expressed variation in the various diseases generating process, so that NGAL becomes the biomarker that detects disease.
The result of study of in recent years U.S.'s clinical chemistry association (AACC) annual meeting is delivered and is learnt that neutrophil leucocyte gelatinase relative carrier lipoprotein may help the clinician to detect the toxicity that the turnover in patients following heart transplantation cyclosporin is induced in the detection urine, and the irreversible injury of regulating ring p0-357 dosage to prevent it that kidney is caused.Chinese patent 200580048175 is studied early stage that matter ephrosis (ATIN) is fallen ill between acute renal failure (ARF), acute tubular necrosis (ATN) and acute tubular, the NGAL level significantly increases in urine and the blood, so NGAL is that the early detection kidney is to the mark of ischemia injury.
At present, detect the NGAL method mainly contain euzymelinked immunosorbent assay (ELISA) (Chinese patent 200810237423.X), based on micro-column method of testing (Chinese patent 200580048175) and the latex immunoturbidimetry (Chinese patent 20101036227.0) of antibody mediated immunity.The euzymelinked immunosorbent assay (ELISA) processing ease is subject to the impact that enzymatic activity changes, and the result is not too accurate; Latex turbidimetry atopic is bad, required reagent more complicated.
Therefore, transmitting light immuno analytical method exploitation NGAL quick detection reagent in the application is applied to the bedside quick diagnosis and has important clinical value.
Summary of the invention
In order to overcome above-mentioned deficiency of the prior art, the utility model provide a kind of based on turn the mensuration NGAL immunochromatography quantification kit of luminescence method, have accurately and rapidly characteristics.
ELISA test strip principle of the present utility model is by a series of finishinges and activation, upper forwarding light particle UCP can combine with bioactive molecule, utilize the UPT biology sensor, the UCP particle that is incorporated in detection zone and the Quality Control district by Immunel response in chromatography process is carried out scanning analysis, realize accurate quantification or qualitative thereby utilize by UPT rapidly and quantitatively testing system.
The technical scheme that provides of the present utility model is:
The upper forwarding light fast quantification kit that a kind of NGAL detects, be included in sample pad, pad, analyzing film and the thieving paper pasted successively on the end liner, be coated with the NGAL antibody I of forwarding light UCP particle marker on the described pad, described analyzing film is provided with parallel detection zone and Quality Control district, described detection zone is coated with NGAL antibody I I or NGAL antigen, and described Quality Control district is coated with sheep anti-mouse igg.
The diameter of described upper forwarding light UCP particle is 50-350nm.
Described NGAL antibody I and NGAL antibody I I are pairing monoclonal antibody or monoclonal antibody-how anti-pairing.
Described NGAL antigen is human neutrophil genatinase associated lipocalin.
The preparation method of the upper forwarding light fast quantification kit that the utility model NGAL detects is:
⑴ pad preparation: it is 2mg/ml that the UCP-NGAL antibody I is diluted to final concentration with the 0.03M PBS damping fluid (containing 2%BSA and 2% trehalose) of pH7.5, is sprayed on the pad 37 ℃ of dry for standby behind the abundant mixing;
⑵ analyzing film preparation: with point sample instrument diverse location difference specking NGAL antibody I I (or NGAL antigen) and sheep anti-mouse igg antibody on analyzing film, as detection zone and Quality Control district, 37 ℃ of dry for standby;
⑶ sample pad preparation: the 0.03M PBS damping fluid (containing 5mg/mlBSA and 0.1%TritonX-100) of sample pad being put into pH 7.2 soaks 1-2h, then 37 ℃ of dry for standby;
⑷ kit assembling: sample pad, pad, analyzing film and thieving paper are sticked on the end liner successively, cut into the specified width, which width test strips and get final product.
Compared with prior art, the utlity model has following useful technique effect:
The utility model detection kit is based on the application of transmitting light technology and immunochromatography technique and be combined in quantitative detection NGAL, so that NGAL detects without background interference, testing result is sensitiveer; Compared with prior art, this kit stability is higher.
Description of drawings
The upper forwarding light fast quantification kit structural representation that Fig. 1 NGAL detects.
Wherein, 1, end liner; 2, sample pad; 3, pad; 4, analyzing film; 5, thieving paper; 6, detection zone; 7, Quality Control district.
Embodiment
Below in conjunction with accompanying drawing the utility model is described in further detail, described is to explanation of the present utility model rather than restriction.
As shown in Figure 1, the upper forwarding light fast quantification kit that a kind of NGAL detects, be included in sample pad 2, pad 3, analyzing film 4 and the thieving paper 5 pasted successively on the end liner 1, be coated with the NGAL antibody I of forwarding light UCP particle marker on the described pad 3, described analyzing film 4 is provided with parallel detection zone 6 and Quality Control district 7, described detection zone 6 is coated with NGAL antibody I I or NGAL antigen, and described Quality Control district 7 is coated with sheep anti-mouse igg.
The diameter of described upper forwarding light UCP particle is 50-350nm.
Described NGAL antibody I and NGAL antibody I I are pairing monoclonal antibody or monoclonal antibody-how anti-pairing.
Described NGAL antigen is human neutrophil genatinase associated lipocalin.
The preparation of the upper forwarding light fast quantification kit that embodiment 1 the utility model NGAL detects
Method 1:
⑴ pad 3 preparations: 50nmUCP particle and NGAL monoclonal antibody I covalent coupling are obtained UCP-NGAL monoclonal antibody I, then using the 0.03M PBS damping fluid (containing 2%BSA and 2% trehalose) of pH7.5 to be diluted to final concentration is 2mg/ml, fully be sprayed on the pad 37 ℃ of dry for standby behind the mixing;
⑵ analyzing film 4 preparation: the solution that NGAL monoclonal antibody II or NGAL polyclonal antibody and rabbit anti-mouse igg antibody is configured to respectively 0.2mg/ml and 0.3mg/ml with the PBS damping fluid of pH 7.2, rule respectively as 7,37 ℃ of dry for standby of detection zone 6 and Quality Control district with the amount of 1 μ l/cm on analyzing film 4 with spray film instrument again;
⑶ sample pad 2 preparations: the 0.03M PBS damping fluid (containing 5mg/mlBSA and 0.1%TritonX-100) of sample pad 2 being put into pH 7.2 soaks 30min, then 37 ℃ of dry for standby;
⑷ kit assembling: sample pad 2, pad 3, analyzing film 4 and thieving paper 5 are sticked on the end liner 1 successively, cut into the specified width, which width test strips and get final product.
Method 2:
⑴ pad 3 preparations: 350nmUCP particle and NGAL monoclonal antibody I covalent coupling are obtained UCP-NGAL monoclonal antibody I, then using the 0.03M PBS damping fluid (containing 2%BSA and 2% trehalose) of pH7.5 to be diluted to final concentration is 2mg/ml, fully be sprayed on the pad 37 ℃ of dry for standby behind the mixing;
⑵ analyzing film 4 preparations: the solution that human neutrophil genatinase associated lipocalin and rabbit anti-mouse igg antibody is configured to respectively 0.2mg/ml and 0.3mg/ml with the PBS damping fluid of pH 7.2, rule respectively as 7,37 ℃ of dry for standby of detection zone 6 and Quality Control district with the amount of 1 μ l/cm on analyzing film 4 with spray film instrument again;
⑶ sample pad 2 preparations: the 0.03M PBS damping fluid (containing 5mg/mlBSA and 0.1%TritonX-100) of sample pad 2 being put into pH 7.2 soaks 30min, then 37 ℃ of dry for standby;
⑷ kit assembling: sample pad 2, pad 3, analyzing film 4 and thieving paper 5 are sticked on the end liner 1 successively, cut into the specified width, which width test strips and get final product.
To be detected urine and drip 100-120 μ l on sample pad 2, solution will dissolve the UCP particle that mark is good on the pad 3, and at analyzing film 4 chromatography occurs, then with the UPT biology sensor test strips is scanned in 5-10min, the content that draws the NGAL in the testing sample according to signal carries out quantitatively or qualitative detection.
Claims (4)
1. the upper forwarding light fast quantification kit that detects of a NGAL, be included in sample pad (2), pad (3), analyzing film (4) and the thieving paper (5) pasted successively on the end liner (1), it is characterized in that being coated with on the described pad (3) the NGAL antibody I of forwarding light UCP particle marker, described analyzing film (4) is provided with parallel detection zone (6) and Quality Control district (7), described detection zone (6) is coated with NGAL antibody I I or NGAL antigen, and described Quality Control district (7) is coated with sheep anti-mouse igg.
2. the upper forwarding light fast quantification kit that detects of described NGAL according to claim 1 is characterized in that the diameter of described upper forwarding light UCP particle is 50-350nm.
3. the upper forwarding light fast quantification kit that detects of described NGAL according to claim 1 is characterized in that described NGAL antibody I and NGAL antibody I I are pairing monoclonal antibody or monoclonal antibody-how anti-pairing.
4. the upper forwarding light fast quantification kit that detects of described NGAL according to claim 1 is characterized in that described NGAL antigen is human neutrophil genatinase associated lipocalin.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201220497401 CN202956384U (en) | 2012-09-26 | 2012-09-26 | Up-converting phosphor rapid ration kit used for neutrophil gelatinase associated lipocalin (NGAL) detection |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201220497401 CN202956384U (en) | 2012-09-26 | 2012-09-26 | Up-converting phosphor rapid ration kit used for neutrophil gelatinase associated lipocalin (NGAL) detection |
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| CN202956384U true CN202956384U (en) | 2013-05-29 |
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| CN 201220497401 Expired - Lifetime CN202956384U (en) | 2012-09-26 | 2012-09-26 | Up-converting phosphor rapid ration kit used for neutrophil gelatinase associated lipocalin (NGAL) detection |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105527439A (en) * | 2015-12-30 | 2016-04-27 | 厦门依柯利斯医疗科技有限公司 | An NGAL time-resolved fluoroimmunoassay nanometer immunochromatographic quantitative detection test paper strip and a preparing method thereof |
| CN105842464A (en) * | 2016-06-12 | 2016-08-10 | 吉林大学 | Device for jointly quantitatively detecting uNGAL (urinary Neutrophil Gelatinase Associated Lipocalin) and uCr (urinary Creatinine) based on up-converting phosphor technology and preparation method of device |
| CN105911274A (en) * | 2016-06-12 | 2016-08-31 | 吉林大学 | Immunochromatography device for synchronously and quantitatively detecting different molecular forms of human neutrophil lipocalin (HNL) and preparation method thereof |
-
2012
- 2012-09-26 CN CN 201220497401 patent/CN202956384U/en not_active Expired - Lifetime
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105527439A (en) * | 2015-12-30 | 2016-04-27 | 厦门依柯利斯医疗科技有限公司 | An NGAL time-resolved fluoroimmunoassay nanometer immunochromatographic quantitative detection test paper strip and a preparing method thereof |
| CN105842464A (en) * | 2016-06-12 | 2016-08-10 | 吉林大学 | Device for jointly quantitatively detecting uNGAL (urinary Neutrophil Gelatinase Associated Lipocalin) and uCr (urinary Creatinine) based on up-converting phosphor technology and preparation method of device |
| CN105911274A (en) * | 2016-06-12 | 2016-08-31 | 吉林大学 | Immunochromatography device for synchronously and quantitatively detecting different molecular forms of human neutrophil lipocalin (HNL) and preparation method thereof |
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| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CX01 | Expiry of patent term |
Granted publication date: 20130529 |
|
| CX01 | Expiry of patent term |