CN206804667U - A kind of multi objective time-resolved fluoroimmunoassay for renal failure Quantitative detection chromatographs kit - Google Patents

Abstract

It the utility model is related to a kind of multi objective time-resolved fluoroimmunoassay for renal failure Quantitative detection and chromatograph kit, including detection card and selectable fluorescence immunoassay quantitative analysis instrument, detection card includes loading pad, pad, detecting pad with detection line and nature controlling line, sample suction pad and bottom plate, Testing index includes kidney injury molecule-1, the gelatinase-associated lipocalin protein of neutrophil leucocyte and RBP ELISA, pad is included with reference to pad body and coated on the time-resolved fluorescence microballoon labelled antibody coating combined on pad body, antibody in the time-resolved fluorescence microballoon labelled antibody coating is the antibody of the index；Changed by the index for detecting Kim 1, NGAL and RBP simultaneously, in time, it is easy, directly perceived, efficiently the generation to renal failure, development synchronize Quantitative Monitoring, family's detection and Site Detection can be realized, required sample size is few, high sensitivity, and detection background signal is low, high specificity, the range of linearity is wide, easy to operate, reliable results.

Description

A kind of multi objective time-resolved fluoroimmunoassay for renal failure Quantitative detection chromatographs Kit

Technical field

It the utility model is related to biotechnology diagnostic field, and in particular to a kind of more fingers for renal failure Quantitative detection Mark time-resolved fluorescence immune chromatography reagent kit.

Background technology

Renal failure is a continuity process, betides diabetes patient, traffic accident injury, patient with severe symptoms, major operation extensively Patient afterwards, it is divided into risk, damage, exhaustion, renal function forfeiture and ESRD by FIFLE standards.In the prior art, it is clinical The biomarker for detecting renal failure is creatinine, urea nitrogen and urine volume.And these traditional index cause because its influence factor is more Specificity is relatively low, causes serious limitation using creatinine etc. as renal failure diagnostic markers be present.

The early stage of renal failure occurs, evolution includes renal damage, detection of glomeruli filtration function declines, the inspection of renal failure process Survey helps to implement protectiveness treatment to patient, prevents the further deterioration of renal function.In view of existing clinic face these Problem, the biomarker of new early stage acute injury of kidney are progressively developed.Including Kim1 (Kim-1), The gelatinase-associated lipocalin protein of neutrophil leucocyte (NGAL) and RBP ELISA（RBP）.

The I type transmembrane protein that Kim1 (Kim-1) is made up of 334 amino acid residues, in normal kidney group Hardly expressed in knitting, be in high expression status in people's proximal tubular epithelial cells after ischemic and injury of kidney occur for kidney.Kidney At least more than 5 times of KIM-1 rises in 1st day after tubule is impaired, and serum creatinine raises with urea nitrogen evening to impaired rear 3rd talent, Therefore Kim-1 is one of most sensitive mark for monitoring early stage renal damage.The extracellular ectodomain of Kim-1 molecules (metalloprotein enzyme hydrolysis takes off, and can be secreted into urine) is acute renal failure (AKI) quantitative marker, in urine KIM-1 detection is not influenceed by urine physics and chemistry material, after openheart surgery several hours will detect that Kim-1 is big in urine Width increases phenomenon.Kim-1 horizontal rises are urinated during renal ischemic, there is higher specificity, for AKI patient, high level The prognosis of Kim-1 indication patients is poor.Clinical experiments have proved that Kim-1 is one of most sensitive mark of early stage renal damage.

The gelatinase-associated lipocalin protein of neutrophil leucocyte (NGAL) is a kind of small-molecular-weight secreted protein, by Kjeldsedn et al. had found in 1993, was the newcomer of Lipocalin families.Under physiological condition, NCAL synthesizes in bone on a small quantity The epithelial cell of multiple systems such as marrow neutrophil leucocyte and renal tubule, inflammatory reaction and malignant tumour can lure that it is being organized more into Largely generated in (such as uterus, prostate, salivary gland, lung, kidney, air flue and alimentary canal epithelium), can reaches in several hours To peak value, produce speed and be significantly faster than that other relevant biomarkers things.NGAL has just expressed in acute tubular damage early stage, Reflect that renal tubular function is disorderly, renal damage is prompted in the rise of its concentration, and the rise of traditional index serum creatinine then needs 24 More than hour.NGAL is also the preferred index thing of current new A KI diagnosis.

RBP ELISA（RBP）It is the small molecular weight protein that a kind of molecular weight is about 21KD, mainly by liver cell Synthesis, is distributed widely in each pseudo body fluid such as blood, cerebrospinal fluid, the RBP in blood often with retinol and transthyretin Ternary complex is combined to form, the RBP of bonding state can not be from glomerular filtration.When kidney filtering function reduces, kidney is small Ball filtration rate and renal blood flow reduce and make various forms of RBP ELISAs in blood（RBP）Store up and show serum RBP Concentration raises.In CGN, serum RBP is good with renal function correlation, can be used as detection of glomeruli filtration function by The sensitive indicator of damage.RBP ELISA in normal human urine（RBP）Content is seldom, about below 0.1ug/min, and its level increases It is considered as a kind of diagnosis index of sensitive proximal tubular damage to add.RBP and urinary albumin, NAG, B2M etc. There is significant positive correlation.And it is sensitive compared with NAG as the index of renal damage to urinate RBP.Under morbid state, urine RBP concentration is again It is 10 times of NAG.Therefore, the measure for urinating RBP is more more valuable than NAG and β 2-MG.When kidney disease or infection etc. cause renal tubule During reabsorption dysfunction, RBP concentration raises in urine, therefore RBP measure is to diagnose the sensitivity of early stage renal damage in urine Index.

The content of the invention

When technical problem to be solved in the utility model is to provide a kind of multi objective for renal failure Quantitative detection Between resolved fluorometric immune chromatography reagent kit, its by simultaneously detect Kim-1, NGAL and RBP index change, can detect exactly The damage of early stage renal tubule, there is the characteristics of high specificity, accuracy is high, bled with one, a drop urine realizes family's detection and existing Field detection, avoids the further generation of renal failure, has high social benefit.

To solve above technical problem, the utility model adopts the following technical scheme that：

Kit include detection card and selectable fluorescence immunoassay quantitative analysis instrument, detection card include loading pad, pad, Detecting pad, sample suction pad and bottom plate with detection line and nature controlling line, it is bright that Testing index includes Kim1, neutrophil leucocyte Glue enzyme correlation lipocalin protein and RBP ELISA, pad are included with reference to pad body and coated on reference to pad body On time-resolved fluorescence microballoon labelled antibody coating, the antibody in the time-resolved fluorescence microballoon labelled antibody coating is described The antibody of index；Detection line has three, and this three detections line is separated from each other and respectively can be with time-resolved fluorescence microballoon mark Remember the coating for matching antibody or antigen and being formed that antibody specificity combines.

Monoclonal antibody corresponding with Kim-1, NGAL, RBP respectively, polyclonal antibody, or phase are coated with detecting pad The nature controlling line that the detection line and mouse IgG antibody that the antigen answered is formed are formed.

The utility model also provides a kind of multi objective time-resolved fluoroimmunoassay for renal failure Quantitative detection and chromatographed The scheme of kit, the kit include detection card and selectable fluorescence immunoassay quantitative analysis instrument, detection card include loading pad, Pad, detecting pad, sample suction pad and the bottom plate with detection line and nature controlling line, it is thin that index includes Kim1, neutral grain The gelatinase-associated lipocalin protein of born of the same parents and RBP ELISA, kit also include fluorescent labeled antibody solution, wherein Antibody in time-resolved fluorescence microballoon labelled antibody coating is the antibody of the index；Detection line has three, this three detections The pairing antibody or antigen shape that line is separated from each other and can respectively specifically bound with time-resolved fluorescence microballoon labelled antibody Into coating.

According to a specific aspect of the present utility model, detection blocks by loading pad, pad, detecting pad and sample suction pad successively Interlaced be pasted onto on bottom plate is formed.

According to a preferred aspect of the present utility model, detection card also includes hemofiltration film or urine filtering film, hemofiltration film or Urine filtering film is covered on loading pad, and loading pad, pad, detecting pad and sample suction pad are interlaced successively to be pasted onto bottom plate On.

According to another preferred aspect of the present utility model, detection card also includes boosting pad, and detects card by loading pad, combination Interlaced be pasted onto on bottom plate is formed successively for pad, boosting pad, detecting pad and sample suction pad.

According to an also preferred aspect of the present utility model, detection card also includes hemofiltration film or urine filtering film and boosting pad, Hemofiltration film or urine filtering film are covered on loading pad, and loading pad, pad, boosting pad, detecting pad and sample suction pad are mutual successively Staggeredly it is pasted onto on bottom plate.

Preferably, the material of boosting pad is glass fibre element film or non-woven fabrics.

According to a preferred aspect of the present utility model, the detecting pad only has one, the three detections line, nature controlling line It is parallel to each other and is successively set on along the length direction of the detecting pad on the detecting pad.

Further, the detection line of close pad is the pairing antibody or antigen of Kim1 in three detections line Coating, the detection line close to sample suction pad is the pairing antibody or antigen coating of RBP ELISA, during middle detection line is The pairing antibody or antigen coating of the property gelatinase-associated lipocalin protein of granulocyte, nature controlling line are located at the close suction of detecting pad The end of sample pad.

According to another preferred aspect of the present utility model, the detecting pad includes three detection point pads, three detections Line is respectively distributed on three detection point pads.Nature controlling line is respectively arranged with each detection point pad.Kim-1, NGAL, RBP are relative The monoclonal antibody answered, polyclonal antibody, or corresponding antigen are coated on three detecting pads respectively, parallel on every detecting pad The antibody such as nature controlling line, coating sheep anti-mouse igg, goat-anti chicken IgY or goat anti-rabbit igg is set, or only pad handled by glass Liquid processing, prepare the Kim-1 monoclonal antibodies comprising fluorescent microsphere mark, the NGAL monoclonal antibodies and glimmering of fluorescent microsphere mark The fluorescent liquid of the RBP monoclonal antibodies of light microballoon mark.

According to another preferred aspect of the present utility model, detection card includes three independent detections point card, loading pad, combines Pad, sample suction pad and bottom plate have three respectively, and each loading pad, pad, detection point pad, sample suction pad and bottom plate are assembled into one Detection point card.

Preferably, nature controlling line includes but is not limited to the antibody such as sheep anti-mouse igg, goat-anti chicken IgY or goat anti-rabbit igg by coating Formed.

Preferably, detecting pad is nitrocellulose filter and is 5~12um of aperture porous spline structure film；Loading pad, combine The material of pad is glass fibre element film or non-woven fabrics, and the material of sample suction pad is absorbent filter.

Fluorescent microsphere TIME RESOLVED TECHNIQUE and immuno-chromatographic assay technology are known, the time described in the utility model point Distinguish that fluorescent microsphere labelled antibody can use method well known in the art to prepare.

Due to the implementation of above technical scheme, the utility model has the following advantages that compared with prior art：

Innovative point of the present utility model is by while detects Kim-1, NGAL and RBP index change, timely, letter Just, intuitively, efficiently the generation to renal failure, development synchronize Quantitative Monitoring, can realize family detection and Site Detection, it is required Sample size is few, high sensitivity, and detection background signal is low, and high specificity, the range of linearity is wide, easy to operate, reliable results.

The utility model detection card will can be only applied to the kit of hospital laboratory originally, by technological innovation, pass through Increase hemofiltration film or urine filtering film, its generation, development for conveniently, simply, being accurately applied to renal failure is carried out together Quantitative Monitoring is walked, suitable for the simple and crude community sanitary institute of family, home for destitute, condition, its application method is also only by patient or family Category acupuncture one is bled（Or directly use urine）To realize self synchronous detection of inflammation, renal failure and heart failure, mitigation goes to hospital to register, It is lined up, chemically examines, taking the complicated processes such as report, mitigates the pressure of public medical mechanism, be relatively beneficial to the daily treatment of patient.

Further, the utility model adds boosting pad in structure, can effectively remove ambient interferences, so as to enter One step improves diagnostic sensitivity height and the degree of accuracy, and boosting pad additionally aids quickening detection sample flow in addition, improves detection efficiency, Shorten detection time.

Brief description of the drawings

Fig. 1 is the main structure diagram blocked according to the detection of embodiment 1 of the present utility model；

Fig. 2 is the main structure diagram blocked according to the detection of embodiment 2 of the present utility model；

Fig. 3 is the dimensional structure diagram blocked according to the detection of embodiment 3 of the present utility model；

Fig. 4 is the main structure diagram according to the detection point card of embodiment 4 of the present utility model（Detection line containing T1）；

Fig. 5 is the main structure diagram according to the detection point card of embodiment 4 of the present utility model（Detection line containing T2）；

Fig. 6 is the main structure diagram according to the detection point card of embodiment 4 of the present utility model（Detection line containing T3）；

Wherein：1,1 ', 1 ", loading pad；2,2 ', 2 ", gold conjugation pad；20,20 ', 20 ", with reference to pad body；21, 21 ', 21 ", colloidal gold labeled monoclonal antibody coating；3rd, detecting pad；30,30 ', 30 ", detection point pad；4,4 ', 4 ", sample suction pad；5,5’, 5 ", bottom plate；6,6 ', 6 ", hemofiltration film or urine filtering film；7,7 ', 7 ", boosting pad；T1, T2, T3, detection line；C, C ', C ", matter Control line.

Embodiment

Embodiment 1

As shown in figure 1, this example provides a kind of multi objective time-resolved fluoroimmunoassay layer for renal failure Quantitative detection Kit is analysed, it includes detection card, fluorescence immunoassay quantitative analysis instrument, and detection card includes loading pad 1, pad 2, has detection line Detecting pad 3, sample suction pad 4 and bottom plate 5, hemofiltration film or urine filtering film 6 and boosting pad 7, hemofiltration film or urine mistake with nature controlling line C Filter membrane 6 is covered on loading pad 1, and loading pad 1, pad 2, detecting pad 3 and sample suction pad 4 are interlaced successively to be pasted onto bottom plate 5 On.Wherein, the material of boosting pad 7 is glass fibre element film or non-woven fabrics.Detecting pad 3 be nitrocellulose filter and be aperture 5~ 12um porous spline structure film；Loading pad 1, the material of pad 2 are glass fibre element film or non-woven fabrics, the material of sample suction pad 4 For absorbent filter.

Further, detection card can detect the gelatinase-associated lipid fortune of Kim1, neutrophil leucocyte simultaneously Three kinds of indexs of albumen and RBP ELISA are carried, pad 2 is included with reference to pad body 20 and coated on combination pad body 20 Time-resolved fluorescence microballoon labelled antibody coating 21, the antibody in the time-resolved fluorescence microballoon labelled antibody coating 21 is institute State the antibody of index.The detecting pad 3 is one, has three detections line T1, T2, T3 thereon, this three detections line T1, T2, T3 phase The mutually coating for matching antibody or antigen and being formed of separation and respectively three kinds marks, the pairing antibody or antigen of three kinds of marks It is able to specifically bind with time-resolved fluorescence microballoon labelled antibody coating.Three detections line T1, T2, T3, nature controlling line C phases It is mutually parallel and set gradually along the length direction of detecting pad 3.Close to the detection line of pad 2 in three detections line T1, T2, T3 T1 is the pairing antibody or antigen coating of Kim1, and the detection line T3 close to sample suction pad 4 is RBP ELISA Antibody or antigen coating are matched, middle detection line T2 is the gelatinase-associated lipocalin protein of neutrophil leucocyte with confrontation Body or antigen coating, nature controlling line C are located at the end of the close sample suction pad 4 of detecting pad 3.Nature controlling line C is by including but unlimited In the formation such as sheep anti-mouse igg, goat-anti chicken IGY or goat anti-rabbit igg.

Each part in this example is so assemblied together, and in hothouse, 20~25 DEG C of temperature, humidity is less than 40%, bottom plate 5 is taken, the middle part that coated detecting pad 3 is placed on to bottom plate 5 is pasted, and pad is cut into suitable width, Boosting pad 7 is overlapped close to detection line side in detecting pad 3, pad 2 is cut into suitable width, is pasted onto on boosting pad 7, Overlapped in the opposite side of pad 2 and paste loading pad 1；On the loading pad 1 that hemofiltration film or urine special filtering film 6 are covered in；Examining Survey the close nature controlling line side overlap joint sample suction pad 4 of pad 3.The bottom plate posted is finally cut into the wide test paper of 3~5mm with cutter Bar, load and detection card is formed in plastic clip.

The detection method of this example kit is as follows（Dry method loading）：

(1) detection reagent and sample are balanced to room temperature, takes out detection card, keep flat；

(2) sample to be tested is drawn, adds in the centrifuge tube of cleaning, is diluted with Sample dilution, is fully mixed；

(3) sample after drawing dilution with liquid-transfering gun is added in the sample aperture of loading pad 1, after 15 minutes, according to pre- The standard curve for being first arranged on time-resolved fluoroimmunoassay quantitative readout instrument is calculated simultaneously Kim-1, NGAL and RBP concentration Show result.

The Cleaning Principle of this example detection card is as follows：

After sample is added to sample well, Kim-1, NGAL or RBP antigen and the fluorescent microsphere mark that contain in sample Kim-1, NGAL or RBP antibody binding, by capillarity, the antibody-antigen immune compound formed is along cellulose nitrate Plain film layer is analysed to detection line, and high specific and height parent then occur with Kim-1, NGAL and RBP antibody being solidificated in detection line With the immune response of property, thus immune complex is enriched with or is trapped in the detection band of chromatographic material, and uncombined exempts from Epidemic disease compound then continues chromatography and captured to nature controlling line by sheep anti-mouse igg antibody.After the completion of reaction, Eu in detection band ^{3 +}Chelating The content of thing has certain corresponding relation with the concentration of target checking matter.If being free of checking matter in testing sample solution, Just there is no Eu in detection band^{3 +}Chelate.It is conventional fluorescent because the fluorescence decay time of lanthanide ion chelate is grown 10³—10⁶Times.When measuring its fluorescence with time-resolved fluorescence instrument, after light-pulse generator excites, a period of time can be postponed again Measure, now the short-half-life fluorescence of other compositions has decayed, and only exists Eu³⁺The specificity fluorescent of label, passes through Eu on ELISA test strip band after measurement immune response ^{3 +}The content of chelate, reference standard concentration curve can quantify The concentration of antigen is corresponded into testing sample.

After reaction terminates, analyzed, according to being set in advance in time-resolved fluoroimmunoassay quantitative readout instrument Standard curve to Kim-1, NGAL and RBP concentration carries out that result is calculated and be shown.

Embodiment 2

As shown in Fig. 2 this example provides a kind of multi objective time-resolved fluoroimmunoassay layer for renal failure Quantitative detection Analyse kit, its substantially with embodiment 1, unlike, its pad does not have time-resolved fluorescence microballoon labelled antibody coating, Kit still further comprises time-resolved fluorescence microballoon labelled antibody solution.

The detection method of this example kit is as follows（Wet method loading）：

(1) detection reagent and sample are balanced to room temperature, takes out detection card, keep flat；

(2) sample to be tested is drawn, is sufficiently mixed with time-resolved fluorescence microballoon labelled antibody solution；

(3) draw mixed liquor with liquid-transfering gun to be added in the sample aperture of loading pad 1, after 15 minutes, according to having pre-set Kim-1, NGAL and RBP concentration are carried out that knot is calculated and be shown in the standard curve of time-resolved fluoroimmunoassay quantitative readout instrument Fruit.

Embodiment 3

As shown in figure 3, this example provides a kind of multi objective time-resolved fluoroimmunoassay layer for renal failure Quantitative detection Analyse kit, its substantially with embodiment 1, unlike, its detecting pad 3 divides pad 30,30 ' including three detections being arranged side by side, 30 ", each detection divides on pad 30,30 ', 30 " forms a detection line and nature controlling line respectively.

Preparation, detection and the Cleaning Principle of this example kit are the same as embodiment 1.

Embodiment 4

As shown in Fig. 4, Fig. 5, Fig. 6, it is glimmering that this example provides a kind of multi objective time resolution for renal failure Quantitative detection Light immune chromatography reagent kit, its substantially with embodiment 1, unlike, detection card includes three independent detections point and blocked, hemofiltration film Or urine filtering film 6,6 ', 6 ", loading pad 1,1 ', 1 ", pad 2,2 ', 2 ", boosting pad 7,7 ', 7 ", detection divide pad 30,30 ', 30 ", sample suction pad 4,4 ', 4 " and bottom plate 5,5 ', 5 " have three respectively, each hemofiltration film or urine filtering film 6,6 ', 6 ", loading Pad 1,1 ', 1 ", pad 2,2 ', 2 ", boosting pad 7,7 ', 7 ", detection divide pad 30,30 ', 30 ", sample suction pad 4,4 ', 4 " and bottom plate 5,5 ', 5 " are assembled into a detection point card, and each detection detected on point card divides on pad 30,30 ', 30 " forms an inspection respectively Survey line and nature controlling line.

Preparation, detection and the Cleaning Principle of this example kit are the same as embodiment 1.

To sum up, innovative point of the present utility model is by while detects Kim-1, NGAL and RBP index change, and When, simplicity, directly perceived, efficiently the generation to renal failure, development synchronize Quantitative Monitoring, family's detection and scene inspection can be realized Survey, required sample size is few, high sensitivity, and detection background signal is low, and high specificity, the range of linearity is wide, easy to operate, as a result may be used Lean on, the accuracy rate of clinical monitoring is up to more than 95%.

The utility model detection card will can be only applied to the kit of hospital laboratory originally, by technological innovation, pass through Increase hemofiltration film or urine filtering film, its generation, development for conveniently, simply, being accurately applied to renal failure is carried out together Quantitative Monitoring is walked, suitable for the simple and crude community sanitary institute of family, home for destitute, condition, its application method is also only by patient or family Category acupuncture one is bled or directly uses urine）To realize that generation, development to renal failure synchronize Quantitative Monitoring, mitigation goes to hospital to hang Number, be lined up, chemically examine, taking the complicated processes such as report, mitigate the pressure of public medical mechanism, be relatively beneficial to the daily treatment of patient.

Further, the utility model adds boosting pad in structure, can effectively remove ambient interferences, so as to enter One step improves diagnostic sensitivity and the degree of accuracy, and boosting pad additionally aids quickening detection sample flow in addition, improves detection efficiency, contracting Short detection time.

The utility model is described in detail above, its object is to allow the personage for being familiar with this art can be much of that Solve content of the present utility model and be carried out, the scope of protection of the utility model can not be limited with this, it is all according to this practicality The equivalent change or modification that new Spirit Essence is made, it should all cover in the scope of protection of the utility model.

Claims (10)

1. a kind of multi objective time-resolved fluoroimmunoassay for renal failure Quantitative detection chromatographs kit, it includes detection card With selectable fluorescence immunoassay quantitative analysis instrument, the detection card includes loading pad（1）, pad（2）, there is detection line and matter Control line（C）Detecting pad（3）, sample suction pad（4）And bottom plate（5）, it is characterised in that：The index include Kim1, in The property gelatinase-associated lipocalin protein of granulocyte and RBP ELISA, the pad（2）Including with reference to pad body （20）With coated on the combination pad body（20）On time-resolved fluorescence microballoon labelled antibody coating（21）, wherein when described Between resolved fluorometric microballoon labelled antibody coating（21）In antibody be the index antibody；The detection line has three, and this three Bar detection line（T1、T2、T3）Be separated from each other and be respectively can be with the time-resolved fluorescence microballoon labelled antibody coating（21） Antibody specificity combine pairing antibody or antigen formed coating.

2. a kind of multi objective time-resolved fluoroimmunoassay for renal failure Quantitative detection chromatographs kit, it includes detection card With selectable fluorescence immunoassay quantitative analysis instrument, the detection card includes loading pad（1）, pad（2）, there is detection line and matter Control line（C）Detecting pad（3）, sample suction pad（4）And bottom plate（5）, it is characterised in that：The index include Kim1, in The property gelatinase-associated lipocalin protein of granulocyte and RBP ELISA, the kit also include fluorescent labeled antibody Solution, wherein the antibody in the time-resolved fluorescence microballoon labelled antibody coating is the antibody of the index；The detection line There are three, this three detections line（T1、T2、T3）It is separated from each other and can be respectively marked with the time-resolved fluorescence microballoon anti- The coating that the pairing antibody or antigen of body specific binding are formed.

3. multi objective time-resolved fluoroimmunoassay according to claim 1 or 2 chromatographs kit, it is characterised in that：It is described Detection card also includes hemofiltration film or urine filtering film（6）, the hemofiltration film or urine filtering film（6）It is covered in the loading pad （1）On, the loading pad（1）, pad（2）, detecting pad（3）And sample suction pad（4）It is interlaced successively to be pasted onto bottom plate（5） On.

4. multi objective time-resolved fluoroimmunoassay according to claim 1 or 2 chromatographs kit, it is characterised in that：It is described Detection card also includes boosting pad（7）, and the detection card is by the loading pad（1）, pad（2）, boosting pad（7）, detecting pad （3）And sample suction pad（4）It is interlaced successively to be pasted onto the bottom plate（5）Upper composition, the boosting pad（7）Material be glass fibers Tie up plain film or non-woven fabrics.

5. multi objective time-resolved fluoroimmunoassay according to claim 1 or 2 chromatographs kit, it is characterised in that：It is described Detection card also includes hemofiltration film or urine filtering film（6）With boosting pad（7）, the hemofiltration film or urine filtering film（6）It is covered in The loading pad（1）On, the loading pad（1）, pad（2）, detecting pad（3）And sample suction pad（4）Interlaced stickup successively In bottom plate（5）On, the boosting pad（7）Material be glass fibre element film or non-woven fabrics.

6. multi objective time-resolved fluoroimmunoassay according to claim 1 or 2 chromatographs kit, it is characterised in that：It is described Detecting pad only has one, the three detections line（T1、T2、T3）, nature controlling line（C）It is parallel to each other and along the detecting pad（3） Length direction be successively set on the detecting pad.

7. multi objective time-resolved fluoroimmunoassay according to claim 6 chromatographs kit, it is characterised in that：Described three Detection line（T1、T2、T3）In close to pad（2）Detection line（T1）Applied for the pairing antibody or antigen of Kim1 Layer, close to sample suction pad（4）Detection line（T3）For the pairing antibody or antigen coating of RBP ELISA, middle detection line （T2）For the pairing antibody or antigen coating of the gelatinase-associated lipocalin protein of neutrophil leucocyte, the nature controlling line（C）Position In the detecting pad（3）The close sample suction pad（4）End.

8. multi objective time-resolved fluoroimmunoassay according to claim 1 or 2 chromatographs kit, it is characterised in that：It is described Detecting pad（3）Including three detection point pads（30,30 ', 30 "）, the three detections line be respectively distributed to it is described three detection point Pad（30,30 ', 30 "）On.

9. multi objective time-resolved fluoroimmunoassay according to claim 8 chromatographs kit, it is characterised in that：The detection Card includes three independent detections point card, the loading pad（1,1 ', 1 "）, pad（2,2 ', 2 "）, sample suction pad（4,4 ', 4 "） And bottom plate（5,5 ', 5 "）There are three respectively, each loading pad（1,1 ', 1 "）, pad（2,2 ', 2 "）, detection point pad（30, 30 ', 30 "）, sample suction pad（4,4 ', 4 "）And bottom plate（5,5 ', 5 "）It is assembled into the detection point card described in one.

10. multi objective time-resolved fluoroimmunoassay according to claim 1 or 2 chromatographs kit, it is characterised in that：It is described Nature controlling line（C）Formed by being coated with sheep anti-mouse igg, goat-anti chicken IgY or goat anti-rabbit igg antibody, the detecting pad（3）It is fine for nitric acid Tie up plain film and for 5~12um of aperture porous spline structure film；The loading pad（1）, pad（2）Material for glass fibre element Film or non-woven fabrics, the sample suction pad（4）Material be absorbent filter.