CN204287201U - Neutrophil leucocyte gelatinase relative carrier lipoprotein immune chromatography detecting test paper strip - Google Patents
Neutrophil leucocyte gelatinase relative carrier lipoprotein immune chromatography detecting test paper strip Download PDFInfo
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- CN204287201U CN204287201U CN201420648456.4U CN201420648456U CN204287201U CN 204287201 U CN204287201 U CN 204287201U CN 201420648456 U CN201420648456 U CN 201420648456U CN 204287201 U CN204287201 U CN 204287201U
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Abstract
The utility model discloses a kind of neutrophil leucocyte gelatinase relative carrier lipoprotein immune chromatography detecting test paper strip, comprise base plate, sample pad, fluorescent marker pad, nitrocellulose filter and adsorptive pads; Described test strips is overlapped successively to be pasted onto on base plate formed by sample pad, fluorescent marker pad, nitrocellulose filter, adsorptive pads.Described fluorescent marker pad contains marked by streptavidin fluorescin and biotin labeled neutrophil leucocyte gelatinase relative carrier lipoprotein monoclonal antibody; Described nitrocellulose membrane there are detection line and nature controlling line.The utility model is compared with the method for detection neutrophil leucocyte gelatinase relative carrier lipoprotein common at present (as: chemoluminescence method/Immune-enhancing effect turbidimetry/colloidal gold method), not only substantially reduce detection time, also improve detection sensitivity simultaneously.
Description
Technical field
The utility model belongs to field of immunodetection, is specifically related to a kind of high sensitivity neutrophil leucocyte gelatinase relative carrier lipoprotein immune chromatography detecting test paper strip.
Background technology
Neutrophil leucocyte gelatinase relative carrier lipoprotein (neutrophil gelatinase associated lipocalin, NGAL), also known as people's ALPHA-2u (lipocalin 2, or bite ferritin (siderocalin), a newcomer in Shi Ren lipocalin protein family Ln2).A kind of specific granule component that NGAL finds when being the research such as Kjcldsen in 1993 neutrophil leucocyte, with Gelatinase B and GELB (man-ix mctalloproteinase-9, MMP-9) closely related.
Under physiological status, the a small amount of NGAL of the secretion such as neutrophil leucocyte, renal cells, pulmonary alveolar macrophage, bronchiolar epithelium mucilage cell, and in the histocyte such as brain and peripheral nerve, colon, uterus and ovary, placenta, thyroid gland, the expression of NGAL is negative.Time acute kidney injury (AKI), blood, urine NGAL concentration can raise rapidly usually; 2h the most obviously (raises tens to hundred times than critical value); serum creatinine (sCr); traditional renal function index such as urine enzyme often will in 24-72h just rear obviously rising, and thus NGAL can be used for the early diagnosis of AKI.NGAL, can early diagnosis acute injury of kidney as a new injury of kidney mark, reflection renal dysfunction degree, plays an important role to the Treatment monitoring of ephrosis and Index for diagnosis.Along with the fast development of the various detection techniques of NGAL, its clinical application research also more and more extensively, deeply.At present because the Object Selection in different result of study, diagnostic criteria, experimental technique design different, be also difficult to carry out lateral comparison.
Biotin-avidin system (biotinavidin system, BAS) is a kind of new bio reaction amplification system.Along with the appearance of various biotin derivative, BAS is widely used in each field of medical science very soon.This system is applied to SABC, enzyme linked immunological, fluorescence immunoassay, in the detection techniques such as radio-immunity, the susceptibility of above technical method, specificity and stability can be improved significantly, make method easier, contribute to clinical quick diagnosis, and be beneficial to and carry out extensive epidemiology survey, become the immunoreactive powerful of research.Because BAS detection system economy is quick, pollute without radiating matter again, do not need complex instrument, fully show the great potential of this system and the possibility of application.
Biotin-avidin system Cleaning Principle: BAS is on the basis of conventional ELISA principle, in conjunction with the height amplification between biotin and Avidin, and a kind of detection system set up.Avidin is a kind of alkaline glycoprotein extracted in ovalbumin, and molecular weight is 68kDa, is made up of 4 subunits, have very high affinity (binding constant is up to 1015M-1) to biology.Biotin is very easily combined with covalent bond with protein (as antibody etc.).Like this, the Avidin molecule combining enzyme and the biotin molecule being combined with specific antibody produce and react, both served multistage amplification, the colour generation due to the catalytic action of enzyme when running into corresponding substrate, reaches the object detecting unknown antigen (or antibody) molecule again.Streptavidin is and the affine a kind of protein have similar biological properties, in Streptavidin molecule the same as Avidin, every bar peptide chain can in conjunction with a biotin, because nearly all material for marking all can combine with Avidin or streptavidin.Utilize biotin-avidin system to develop neutrophil leucocyte gelatinase relative carrier lipoprotein detection examination fast diagnosis reagent and there is no report.
Utility model content
The purpose of this utility model, be to provide a kind of neutrophil leucocyte gelatinase relative carrier lipoprotein immune chromatography detecting test paper strip, it is based on double antibody sandwich method, fluorescence immunoassay lateral chromatography and biotin-Streptavidin amplification system, provides high sensitivity neutrophil leucocyte gelatinase relative carrier lipoprotein immune chromatography detecting test paper strip.When using the utility model, improve detection sensitivity and detect stability, reducing non-specific binding, be not more than 10 minutes detection time, substantially increase diagnosis efficiency.
The solution that the utility model solves its technical matters is: neutrophil leucocyte gelatinase relative carrier lipoprotein immune chromatography detecting test paper strip, it comprises base plate, described base plate is provided with sample pad successively, fluorescent marker pad, nitrocellulose filter and adsorptive pads, described adsorptive pads and fluorescent marker pad are overlapping to be respectively pressed in behind the two ends of nitrocellulose filter in the formation detection zone, surface of nitrocellulose filter, described sample pad is overlapping to be pressed on fluorescent marker pad, described fluorescent marker pad is fixed with the fluorescin of biotin labeled neutrophil leucocyte gelatinase relative carrier lipoprotein monoclonal antibody and marked by streptavidin, nitrocellulose filter in described detection zone is fixed with the nature controlling line of detection line and the sheep anti-mouse igg polyclonal antibody formation identifying that the monoclonal antibody of neutrophil leucocyte gelatinase relative carrier lipoprotein another one epi-position is formed.
As the further improvement of technique scheme, described fluorescent marker pad comprises the first stacked fluorescent marker pad and the second fluorescent marker pad, one end pad of described first fluorescent marker pad in the below of sample pad, described second overlapping one end being pressed in nitrocellulose filter of fluorescent marker pad.
As the further improvement of technique scheme, in described first fluorescent marker pad, the one end of inserting below sample pad is provided with positioning strip, and the lower surface of described sample pad is provided with the locating slot that can hold positioning strip.
As the further improvement of technique scheme, also comprise and getting stuck, in described base plate, sample pad, fluorescent marker pad, nitrocellulose filter and adsorptive pads are all placed in and get stuck, the position described shell face of getting stuck corresponding to nitrocellulose membrane is provided with view window, and position shell face of getting stuck corresponding to sample pad is provided with well.
As the further improvement of technique scheme, described view window is slot.
As the further improvement of technique scheme, described fluorescin can be the one in green fluorescent protein, phycobniliprotein.
As the further improvement of technique scheme, described sample pad is glass fibre component dry after surfactant damping fluid immersion treatment.
As the further improvement of technique scheme, described base plate is polystyrene component or tygon component.
As the further improvement of technique scheme, described in get stuck and comprise plastics upper casing and plastics lower casing, described plastics upper casing fastens to be formed after on plastics lower casing and gets stuck.
Compared with prior art, tool has the following advantages the utility model:
(1) the utility model is using fluorescin as label, and this label has good stability, and is conducive to improving detecting stability.
(2) the utility model is by utilizing " Streptavidin-biotin amplification system ", improves detection sensitivity, reduces non-specific binding, is conducive to improving kit performance.
(3) detection time utilizing the utility model to carry out Procalcitonin is not more than 10 minutes, and the detection range of linearity is 0.10ng/ml ~ 100.00ng/ml, drastically increases detection efficiency.
(4) the utility model carries out interpretation by fluorescence immunity analyzer to result, can realize robotization, reduces the impact of subjective factor, provides convenient, quick, reliable diagnostic result.
(5) novel the getting stuck of this use is provided with the well that sample enters and the view window supplying observed result, according to analytical instrument result of determination, accurately and reliably.
(6) the utility model is easy to make, and volume is little, be easy to carry.
(7) the utility model testing cost is lower.
(8) the utility model can be mass, and is applicable to clinical quick diagnosis and on-the-spot quick diagnosis; Be easy to preserve, be conducive to grass-roots unit and promote.
Accompanying drawing explanation
In order to be illustrated more clearly in the technical scheme in the utility model embodiment, below the accompanying drawing used required in describing embodiment is briefly described.Obviously, described accompanying drawing is a part of embodiment of the present utility model, instead of whole embodiment, and those skilled in the art, under the prerequisite not paying creative work, can also obtain other design proposals and accompanying drawing according to these accompanying drawings.
Fig. 1 is the structural representation of the utility model embodiment one;
Fig. 2 is the structural representation of the utility model embodiment two;
Fig. 3 is the structural representation of the utility model embodiment three;
Fig. 4 is the structural representation of the utility model embodiment four.
Embodiment
Be clearly and completely described below with reference to embodiment and the accompanying drawing technique effect to design of the present utility model, concrete structure and generation, to understand the purpose of this utility model, characteristic sum effect fully.Obviously; described embodiment is a part of embodiment of the present utility model, instead of whole embodiment, based on embodiment of the present utility model; other embodiments that those skilled in the art obtains under the prerequisite not paying creative work, all belong to the scope of the utility model protection.In addition, all connection/annexations mentioned in literary composition, not singly refer to that component directly connects, and refer to and according to concrete performance, can connect auxiliary by adding or reducing, and form more excellent draw bail.
With reference to Fig. 1, neutrophil leucocyte gelatinase relative carrier lipoprotein immune chromatography detecting test paper strip, it comprises base plate 5, described base plate 5 is provided with sample pad 1 successively, fluorescent marker pad 2, nitrocellulose filter 3 and adsorptive pads 4, described adsorptive pads 4 and fluorescent marker pad 2 are overlapping to be respectively pressed in behind the two ends of nitrocellulose filter 3 in the formation detection zone, surface of nitrocellulose filter 3, fluorescent marker pad 2 is glass fibre membrane, described sample pad 1 is overlapping to be pressed on fluorescent marker pad 2, the fluorescin described fluorescent marker pad 2 being fixed with biotin labeled neutrophil leucocyte gelatinase relative carrier lipoprotein monoclonal antibody (concentration 0.3 ~ 1.5mg/mL) and marked by streptavidin (or Avidin) (uses excitation wavelength 530nm, wavelength of transmitted light 570nm, concentration 0.1 ~ 1.0mg/mL), nitrocellulose filter 3 in described detection zone is fixed with the nature controlling line C (concentration 0.2 ~ 2.0mg/mL) of detection line T (concentration 0.5 ~ 3mg/mL) and the sheep anti-mouse igg polyclonal antibody formation identifying that the monoclonal antibody of neutrophil leucocyte gelatinase relative carrier lipoprotein another one epi-position is formed, nature controlling line C is used for the validity of test strip.
The fluorescin that biotin labeled neutrophil leucocyte gelatinase relative carrier lipoprotein monoclonal antibody and Streptavidin (or Avidin) mark is fixed on fluorescent marker pad 2, the monoclonal antibody and sheep anti-mouse igg polyclonal antibody that have identification neutrophil leucocyte gelatinase relative carrier lipoprotein another one epi-position are fixed on nitrocellulose filter as detection line T and nature controlling line C.When testing sample is added to after in sample pad 1, moved forward by chromatography effect, in sample, on neutrophil leucocyte gelatinase relative carrier lipoprotein and fluorescent marker pad 2, neutrophil leucocyte gelatinase relative carrier lipoprotein antibody (Mab-NGAL*Fluoro) of combined with fluorescent label (fluorescin) reacts and forms compound NGAL-Mab-NGAL*Fluoro, compound is reacted when continuing to be advanced past NGAL antibody (detection line) nitrocellulose membrane wrapping quilt under chromatography effect, NGAL antibody capture formation compound (Mab-NGAL-NGAL-Mab-NGAL*Fluoro) (detection line) that reaction compound is coated, the reaction signal of detection line is read by fluorescence immunity analyzer, under the effect of excitation source, fluorescent material launches the fluorescence signal of specific wavelength, fluorescence immunity analyzer captures fluorescence signal, the typical curve transformed by signal and set is converted into quantitative value automatically, calculate the concentration of neutrophil leucocyte gelatinase relative carrier lipoprotein in sample, obtain neutrophil leucocyte gelatinase relative carrier lipoprotein testing result.
As the further improvement of technique scheme, with reference to Fig. 3, described fluorescent marker pad 2 comprises the first stacked fluorescent marker pad 20 and the second fluorescent marker pad 21, one end pad of described first fluorescent marker pad 20 in the below of sample pad 1, described second overlapping one end being pressed in nitrocellulose filter 3 of fluorescent marker pad 21.The layering of fluorescent marker pad 2 is arranged, and is convenient to fluorescent marker pad 2 to be arranged between sample pad 1 and nitrocellulose filter 3.
As the further improvement of technique scheme, be provided with positioning strip 22 with reference to one end of inserting below sample pad in the first fluorescent marker pad 20 described in Fig. 3, the lower surface of described sample pad 1 is provided with the locating slot that can hold positioning strip 22.The installation be convenient between fluorescent marker pad 2 and sample pad 1 by positioning strip 22 is fixed.
As the further improvement of technique scheme, with reference to Fig. 2 and Fig. 4, also comprise and getting stuck, described base plate 5, sample pad 1, fluorescent marker pad 2, nitrocellulose filter 3 and adsorptive pads 4 are all placed in and get stuck in 6, the position described 6 shell faces of getting stuck corresponding to nitrocellulose membrane 3 is provided with view window 8, and the position 6 shell faces of getting stuck corresponding to sample pad 1 is provided with well 7.
As the further improvement of technique scheme, described view window 8 is slot.
As the further improvement of technique scheme, described fluorescin can be the one in green fluorescent protein, phycobniliprotein.
As the further improvement of technique scheme, described sample pad 1 is glass fibre component dry after surfactant damping fluid immersion treatment.Sample pad is made up of glass fibre, through surfactant damping fluid immersion treatment, uses after dry.The cutting width of described sample pad 1 is 0.3 ~ 0.5cm.
As the further improvement of technique scheme, described base plate 5 is polystyrene component or tygon component.
As the further improvement of technique scheme, described in get stuck and 6 comprise plastics upper casing 60 and plastics lower casing 61, described plastics upper casing 61 fastens to be formed after on plastics lower casing 61 and gets stuck 6.
More than that better embodiment of the present utility model is illustrated, but the invention is not limited to described embodiment, those of ordinary skill in the art also can make all equivalent modifications or replacement under the prerequisite without prejudice to the utility model spirit, and these equivalent modification or replacement are all included in the application's claim limited range.
Claims (9)
1. neutrophil leucocyte gelatinase relative carrier lipoprotein immune chromatography detecting test paper strip, it is characterized in that: it comprises base plate, described base plate is provided with sample pad successively, fluorescent marker pad, nitrocellulose filter and adsorptive pads, described adsorptive pads and fluorescent marker pad are overlapping to be respectively pressed in behind the two ends of nitrocellulose filter in the formation detection zone, surface of nitrocellulose filter, described sample pad is overlapping to be pressed on fluorescent marker pad, described fluorescent marker pad is fixed with the fluorescin of biotin labeled neutrophil leucocyte gelatinase relative carrier lipoprotein monoclonal antibody and marked by streptavidin, nitrocellulose filter in described detection zone is fixed with the nature controlling line of detection line and the sheep anti-mouse igg polyclonal antibody formation identifying that the monoclonal antibody of neutrophil leucocyte gelatinase relative carrier lipoprotein another one epi-position is formed.
2. neutrophil leucocyte gelatinase relative carrier lipoprotein immune chromatography detecting test paper strip according to claim 1, it is characterized in that: described fluorescent marker pad comprises the first stacked fluorescent marker pad and the second fluorescent marker pad, one end pad of described first fluorescent marker pad in the below of sample pad, described second overlapping one end being pressed in nitrocellulose filter of fluorescent marker pad.
3. neutrophil leucocyte gelatinase relative carrier lipoprotein immune chromatography detecting test paper strip according to claim 2, it is characterized in that: in described first fluorescent marker pad, the one end of inserting below sample pad is provided with positioning strip, and the lower surface of described sample pad is provided with the locating slot that can hold positioning strip.
4. the neutrophil leucocyte gelatinase relative carrier lipoprotein immune chromatography detecting test paper strip according to any one of claims 1 to 3, it is characterized in that: also comprise and getting stuck, in described base plate, sample pad, fluorescent marker pad, nitrocellulose filter and adsorptive pads are all placed in and get stuck, the position described shell face of getting stuck corresponding to nitrocellulose membrane is provided with view window, and position shell face of getting stuck corresponding to sample pad is provided with well.
5. neutrophil leucocyte gelatinase relative carrier lipoprotein immune chromatography detecting test paper strip according to claim 4, is characterized in that: described view window is slot.
6. neutrophil leucocyte gelatinase relative carrier lipoprotein immune chromatography detecting test paper strip according to claim 4, is characterized in that: described fluorescin can be the one in green fluorescent protein, phycobniliprotein.
7. neutrophil leucocyte gelatinase relative carrier lipoprotein immune chromatography detecting test paper strip according to claim 4, is characterized in that: described sample pad is glass fibre component dry after surfactant damping fluid immersion treatment.
8. neutrophil leucocyte gelatinase relative carrier lipoprotein immune chromatography detecting test paper strip according to claim 4, is characterized in that: described base plate is polystyrene component or tygon component.
9. neutrophil leucocyte gelatinase relative carrier lipoprotein immune chromatography detecting test paper strip according to claim 4, is characterized in that: described in get stuck and comprise plastics upper casing and plastics lower casing, described plastics upper casing fastens to be formed after on plastics lower casing and gets stuck.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2019169309A1 (en) * | 2018-03-02 | 2019-09-06 | Berger Russell Jay | Methods, apparatuses and kits for rapid testing of traumatic brain injuries |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2019169309A1 (en) * | 2018-03-02 | 2019-09-06 | Berger Russell Jay | Methods, apparatuses and kits for rapid testing of traumatic brain injuries |
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