CN202404101U

CN202404101U - Reagent kit

Info

Publication number: CN202404101U
Application number: CN2011203474478U
Authority: CN
Inventors: 张玥; 包骏
Original assignee: Shanghai Kexin Biotech Co Ltd
Current assignee: Shanghai Kexin Biotech Co Ltd
Priority date: 2011-09-16
Filing date: 2011-09-16
Publication date: 2012-08-29
Anticipated expiration: 2021-09-16

Abstract

The utility model provides a reagent kit, which comprises at least two independent test strips, wherein each testing strip comprises a sample pad, a combining pad, a detecting membrane and an absorbent pad which are in lap joint sequentially. A detecting area and a quality control area which are separated from each other are arranged on the detecting membrane, the detecting area is enveloped with detecting protein, and the quality control area is enveloped with quality control protein. The reagent kit can be used for detection of existence of AMA-M2 (anti-mitochondrial antibody-M2), SP100 antibodies, GP210 (nuclear pore glycoprotein 210) antibodies, SLA (soluble liver antigen) antibodies or LKM-1 (liver/kidney microsomal-1) antibodies in detecting samples and auxiliary diagnosis of autoimmune liver diseases.

Description

Kit

Technical field

The utility model relates to a kind of diagnostic kit.More specifically; The utility model relates to the diagnostic kit that comprises at least two test strip; Said at least two test strips are coated with antigen respectively; The AMA-M2 antibody, SP100 antibody, GP210 antibody, SLA antibody or the LKM-1 antibody that are used for external qualitative detection human blood, the auxiliary diagnosis of autoimmune hepatopathy.

Background technology

Background

Hepatitis is one of common disease that influences human health, and chronic hepatitis is bigger to patient and social influence.Abroad reported in succession in recent years autoimmune liver disease, particularly oneself immunity hepatitis (autoimmune hepatitis, AIH) and PBC (primary biliary cirrhosis, PBC), its incidence of disease all has the trend of obvious rising.

AIH is the hepar damnification that is caused by autoimmune response, shows as chronic gradual hepatitis, and until cirrhosis, serology shows as high gamma globulin and characteristic autoantibody etc.Because the detection index of AIH is often overlapping with the indexs of other autoimmune liver diseases, so to agnogenic hepatopath's examination AIH the time, be necessary to get rid of the interference of other diseases.For example, find that the overlapping of AIH and PBC accounts for 8%, account for 6% with primary sclerotic cholangitis is overlapping, account for 11% with slow virus type hepatitis is overlapping, account for 10% with autoimmune cholangitis is overlapping, the slow type hepatitis of other unknown causes accounts for 13%.Wherein the incidence of disease of PBC is higher, therefore, is necessary in examination AIH, to get rid of the interference of PBC.

PBC is that little and median size bile duct is master's an organ specificity autoimmune liver disease in the liver to destroy, and shows as hepatic portal arteries and veins district inflammation and causes fiberization, cirrhosis even functional depletion.PBC patient is the women more than 90%, and the age is generally between 30 to 65 years old, and is in rising trend with the age growth phase incidence of disease.Past, clear and definite PBC method of diagnosing was the liver puncture biopsy, but this traumatic detection means has limited its application.

Up to now, the diagnostic means of autoimmune disease mainly is to combine on the clinical basis, relies on the laboratory and detects the autoantibody that one or more height are tired.In some autoimmune disease diagnostic criteria that internal authority mechanism formulates, just comprised autoantibody; Like PBC (Primary biliary cirrhosis; PBC); In the PBC diagnostic routine that U.S. hepatopathy association (AASLD) delivered in 2000, (AMA) classifies one of diagnostic criteria as with AMA.There are 9 kinds of autoantigens (M1～9) on the mitochondrial membrane, the highest to susceptibility and the specificity of the corresponding AMA of M2 hypotype (AMA-M2) diagnosis PBC wherein.And to the diagnosis of oneself immunity hepatitis (AIH), even can be according to the difference of autoantibody among the patients serum, with its somatotype.Certainly, more than various autoantibodies all have its limitation in the medical diagnosis on disease process, cannot treat different things as the same.The PBC patient negative if any some M2 just can't be diagnosed and treated timely.In recent years detect the autoantibody of some specific types in the existing body that much is reported in the negative PBC patient of M2, the diagnosis of PBC had high degree of specificity, tool clinical value be SP100 antibody and GP210 antibody.SP100 antibody can be detected in PBC patient's body of about 31%, owing to can not in other various types of autoimmune liver diseases, detect this antibody, therefore puts the mark that is regarded as PBC.Though GP210 antibody only is detected in PBC patient's body of about 10%; But the PBC negative to M2 has higher diagnostic value; Can in 21% M2 negative patient body, find, therefore unite SP100 and GP210 antibody index, improve individual event M2 and detected diagnosis PBC.SLA antibody has high degree of specificity to III systemic autoimmune hepatitis, and its positive rate accounts for 11%～32.5% of oneself immunity hepatitis, do not find as yet so far in other diseases or normal population, to have SLA antibody, thereby diagnostic value is very high.LKM antibody (anti-liver/kidney microsomal antibody) can react with cytoplasm of liver, near-end renal tubule, through identifying LKM antibody 3 kinds of hypotypes is arranged, and LKM-1 is the serologic marker of II type oneself immunity hepatitis.

Also do not have a kind of diagnostic device at present, can carry out convenient, fast and examination accurately to AIH and PBC simultaneously.And to the examination of these the two kinds of diseases accuracy to clinical diagnosis, and the successive treatment Scheme Selection all has great importance.In clinical diagnosis; The method of the detection autoantibody that present laboratory is commonly used mainly contains enzyme-linked immunosorbent test (enzyme linked immunosorbent assay; ELISA), IIF (indirect immunoflurescence; IFF), Western blot (immunoblotting test, IBT), linear immunoassay (Line immuno assay, LIA) etc.Though Western blot combines the high resolution of SDS-PAGE and the high specific and the susceptibility of ELISA method, the operation relative complex, and reagent has stronger toxicity and contaminative.IIF can be made semiquantitative determination, but the fluorescence microscope result need be arranged, and the objectivity that the result judges is not enough, the standardization difficulty, and the technical program more complicated also is not suitable for the detection of high flux sample.Linear immunoassay is mainly used in the examination of big type of disease, and specific aim is relatively poor relatively, also is not suitable for the detection of high flux sample simultaneously.The ELISA method detects and can be used for high flux sample mensuration, and sensitivity is also higher, at present by extensive approval; But operate more loaded down with trivial detailsly, need repeatedly application of sample and washing, and be subject to the influence of temperature and incubation conditions; In specialized laboratory, operate, bring inconvenience to experiment.Therefore, this area presses for provides a kind of diagnostic device that in clinical, can detect rapidly and accurately with examination AIH and PBC, the demand that to satisfy the needs of clinical diagnosis, realize detecting immediately, bedside detects.

Summary of the invention

The utility model provides a kind of diagnostic kit that is used for the diagnosis of autoimmune hepatopathy, and it comprises at least two test strips, and wherein said test strips comprises sample pad, pad, detection film and the adsorptive pads of overlap joint successively, and supporting baseplate.Wherein, be coated with on the said pad can with the albumen of the metallic colloid mark of people's antibodies; Be coated with a kind of detection Western blot on the detection zone of said detection film, be coated with a special quality control Western blot in the Quality Control district of said detection film; Said detection Western blot and said Quality Control Western blot on the detection film of said test strips are separated from one another, and said detection Western blot more is close to pad than said Quality Control Western blot.

In some embodiments; Said sample pad is to be processed by spun glass, polyester film, cellulose filter paper or nonwoven fabrics; Said pad is to be processed by spun glass, polyester film, cellulose filter paper or nonwoven fabrics; With said detection film be to process by nitrocellulose filter, and said supporting baseplate by PVC plate or other hard ground and the material that does not absorb water process.

In some embodiments, the shape of said detection Western blot and/or said Quality Control Western blot is respectively mottled or ribbon.In some embodiments, said detection Western blot and said Quality Control Western blot all are mottled.In some embodiments, said detection Western blot and said Quality Control Western blot all are ribbon.

In some embodiments; Detection Western blot on each said test strips is a kind of material that is selected from down group: AMA-M2 Western blot, SP100 Western blot, GP210 Western blot, SLA Western blot and LKM-1 Western blot, the detection Western blot that encapsulates on the different test strips is inequality.

In some embodiments; Said diagnostic kit comprises two said test strips; Detection Western blot on each said test strips is a kind of material that is selected from down group: AMA-M2 Western blot, SP100 Western blot, GP210 Western blot, SLA Western blot and LKM-1 Western blot, the detection Western blot that encapsulates on the different test strips is inequality.

In some embodiments; Said diagnostic kit comprises three said test strips; Detection Western blot on each said test strips is a kind of material that is selected from down group: AMA-M2 Western blot, SP100 Western blot, GP210 Western blot, SLA Western blot and LKM-1 Western blot, the detection Western blot that encapsulates on the different test strips is inequality.

In some embodiments; Said diagnostic kit comprises four said test strips; Detection Western blot on each said test strips is a kind of material that is selected from down group: AMA-M2 Western blot, SP100 Western blot, GP210 Western blot, SLA Western blot and LKM-1 Western blot, the detection Western blot that encapsulates on the different test strips is inequality.

In some embodiments; Said diagnostic kit comprises five said test strips; Detection Western blot on each said test strips is a kind of material that is selected from down group: AMA-M2 Western blot, SP100 Western blot, GP210 Western blot, SLA Western blot and LKM-1 Western blot, the detection Western blot that encapsulates on the different test strips is inequality.

In some embodiments, said Quality Control albumen be can with the antibody or the albumen of antibodies in the sample, perhaps can with the protein bound antibody or the albumen of said colloid gold label.In some embodiments, also be coated with the albumen of second kind of colloid gold label on the pad of said test strips, and said Quality Control Western blot can combine the albumen of said second kind of colloid gold label.

In some embodiments, said diagnostic kit further comprises at least one firm banking, is fixed with said at least two test strips on said at least one firm banking.In some embodiments, be fixed with two to five said test strips on said at least one firm banking, and said test strips is separate.

In some embodiments, the plate face of said firm banking is provided with projection, and wherein said test strips is placed in the accommodation section of being defined by said projection, can hold single test strips.In some embodiments, the plate face of said firm banking is provided with projection, and said projection defines at least two accommodation sections that can hold single test strips.

In some embodiments, said diagnostic kit comprises two or more said firm bankings, wherein is fixed with at least one said test strips respectively on each said firm banking.

In some embodiments; Said diagnostic kit also comprises and said firm banking and the matching used loam cake of said test strips; Each said loam cake has application of sample mouth and watch window; Said application of sample mouth is positioned at the said sample pad top of said test strips, and said watch window is positioned at the said detection film top on the said test strips.

In some embodiments, said at least two test strips are parallel to each other.In some embodiments, the detection Western blot that encapsulates on the detection film of said test strips is positioned on the same horizontal line basically, and the Quality Control Western blot that encapsulates on the detection film of said test strips is positioned on the same horizontal line basically.

In some embodiments, said diagnostic kit comprises that also one or more are selected from down the ingredient of group: firm banking, loam cake, drying agent, dropper, instructions, color label, sample collection tube and sample diluting liquid.

Description of drawings

Fig. 1 is the section exemplary plot of single test strips, wherein 1 indicates sample pad, 2 indication pads, and 3 indications detect films, 4 indication adsorptive pads, 5 indicate detection zones, 6 indication Quality Control districts, 7 indication supporting baseplates.

Fig. 2 is the decomposition exemplary plot of single test strips, wherein the represented same Fig. 1 of structure of 1-7.

Fig. 3 is the exemplary plot that the firm banking that is provided with projection of single test strips can be set, and wherein (a) is vertical view, (b) is cut-open view.Wherein 8 indicate firm bankings, 9 indication projections, 20 indication accommodation sections.

Fig. 4 is the example top view of the loam cake that can cooperate with firm banking, 10 indication loam cakes wherein, 11 indication application of sample mouths, 12 indication watch windows.

Fig. 5 is the firm banking and the loam cake that the are provided with test strips structural representation when cooperating, and wherein (a) is vertical view, (b) is cut-open view, wherein the represented same Fig. 1 of structure, Fig. 2 and the Fig. 4 of 1-12.

Fig. 6 is for being provided with the firm banking that is provided with projection of two test strips and the exemplary plot of the loam cake that can cooperate with it, and wherein (a) is the example top view of firm banking, (b) is the example cut-open view of firm banking, (c) is the example top view of loam cake.Wherein 7 ' indicate firm banking, 8 ' indication projection, 9 ' indication loam cake, 10 ' indication application of sample mouth, 11 ' indication watch window, 20 indication accommodation sections.

Fig. 7 is for being provided with the firm banking that is provided with projection of three test strips and the exemplary plot of the loam cake that can cooperate with it, and wherein (a) is the example top view of firm banking, (b) is the example cut-open view of firm banking, (c) is the example top view of loam cake.Wherein 7 " indication firm banking, 8 " the indication projection, 9 " the indication loam cake, 10 " indication application of sample mouth, 11 " the indication watch window, 20 indication accommodation sections.

Fig. 8 wherein 13 indicates the dark color band that occurs in the detection zones for to detect the testing result synoptic diagram that obtains with test strips, the dark color band that occurs in the 14 indication Quality Control districts.

Embodiment

The utility model provides the diagnostic kit that contains test strips that is used for the diagnosis of autoimmune hepatopathy, and said test strips can be used in the chromatography that carries out sample, and the analyte that exists in can test sample.In some embodiments, said sample can be people's body fluid sample, blood for example, blood plasma etc.In some embodiments, the analyte that exists in the said sample is one or more people's antibody, for example, and AMA-M2 antibody, SP100 antibody, GP210 antibody, SLA antibody or LKM-1 antibody.Above-mentioned these people's antibody are proved to be relevant with autoimmune liver disease, through detecting in blood or the blood plasma whether have one or more these antibody, and can the auxiliary diagnosis of autoimmune hepatopathy.These seized antibody can specificity combine its corresponding antigen, therefore, with the antigen coated detection zone in said test strips of these antibody, can detect corresponding with it seized antibody.Listed the example of the antigen that can combine in the table 1 with AMA-M2 antibody, SP100 antibody, GP210 antibody, SLA antibody or LKM-1 antibody specificity, and the disease that is associated.

Table 1

The diagnostic kit that the application provides comprises at least two test strips 100; Wherein each test strips 100 all comprises sample pad 1, pad 2, detection film 3 and the adsorptive pads 4 of overlap joint successively; And supporting baseplate 7; Wherein, be coated with on the pad of said test strips can with the albumen of the metallic colloid mark of people's antibodies; Be coated with a kind of detection Western blot on the detection zone of the detection film of said test strips, be coated with a special quality control Western blot in the Quality Control district of said detection film; Said detection Western blot and said Quality Control Western blot on the detection film of said test strips are separated from one another, and said detection Western blot more is close to pad than said Quality Control Western blot.In some embodiments, each test strips 100 in the diagnostic kit that the application provides is separate, each other mutually not the chromatographic flow and result's demonstration of sample during Interference Detection.Sample pad 1 is the zone that sample adds.Be coated with the metallic colloid label on the pad 2, the zone that to be sample combine or mix with the metallic colloid label.Detect film 3 and be provided with detection zone separated from one another 5 and Quality Control district 6; And said detection zone 5 than said Quality Control district 6 more near pad 2; Wherein said detection zone 5 is coated with the detection Western blot; Be used to combine antibody to be measured, said Quality Control district 6 is coated with the Quality Control Western blot and is used to combine the Quality Control tester.Adsorptive pads 4 is used to absorb moisture content so that sample moves towards adsorptive pads 4 directions.Supporting baseplate 7 is used to support sample pad 1, pad 2, detection film 3 and the adsorptive pads 4 of overlap joint successively.The concrete structure synoptic diagram of single test strips 100 is seen Fig. 1 and Fig. 2.

As shown in Figure 1, in the single test strips 100, sample pad 1, pad 2, detection film 3 and adsorptive pads 4 from left to right overlap according to this order successively from beginning to end." overlap joint " is meant, the head and the tail of two adjacent assemblies are overlapped, and form the syndeton that can allow fluid sample to move at overlapping.For example, can partly form overlay structure comparatively closely, make sample to move on the downstream components through this overlay structure from the afterbody of upstream component at the head and the tail of two adjacent assemblies through pressing operation.Upstream component can overlap the top of downstream components, also can overlap the below.When detecting; Can be with the sample of liquid condition (like blood; Blood plasma etc.) application of sample is on sample pad 1, and the sample of this liquid state state can pass through water sorption, moves to the pad 2 that overlaps with it along sample pad 1; Move to the detection film 3 that overlaps with it along pad 2 again after touching pad 2, touch detection film 3 and move to the adsorptive pads 4 that overlaps with it along detecting film 3 more afterwards.Like this mobile is commonly referred to the chromatography effect.After sample was added on the sample pad 1, material wherein moved to pad 2 successively and detects on the film 3 through the chromatography effect.When sample moves to pad 2; Detection antibody wherein; Like people's antibody, can combine with the albumen of the metallic colloid mark that encapsulates on the pad 2, contain then from pad 2 and combine or the sample of the material that mixes continues to move to and detects on the film 3; And with detect film 3 on the detection Western blot that encapsulates combine and be trapped, thereby make the analyte in the sample be able to be detected.

In some embodiments, be coated with on the pad 2 of test strips 100 can with first kind of albumen of the metallic colloid substance markers of people's antibodies.In some embodiments, said first kind of albumen can be anti-human IgG antibody, for example, the goat anti-human igg antibody, rabbit anti-human igg's antibody, the anti-human IgG antibody of pig, or mouse-anti is gone into IgG antibody etc.In some embodiments, said first kind of albumen can be staphylococcal protein A or streptococcal protein G.

First kind of albumen of said metallic colloid mark can be through hybrid metal colloid thing and albumen make under proper condition.In some embodiments, can use suitable damping fluid to add in the metallic colloid thing, be adjusted to suitable pH, add albumen more by a certain percentage, mixing makes the albumen of metallic colloid substance markers.Suitable damping fluid comprises, for example, sal tartari damping fluid, borate buffer, phosphate buffer, carbonic acid buffer etc., suitable buffer concentration can be 0.1～0.5M.Suitable pH can be 5-10,6-10,7-10, or 8-9.The metallic colloid thing can make by means commonly known in the art, for example, makes with containing the reaction of metal ion solution and reductive agent.For example; To contain metal ion solution under effects such as reductive agent such as white phosphorus, ascorbic acid, sodium citrate, tannic acid; Aggregate into a certain size metallic particles, and, form electronegative hydrophobic colloid solution because electrostatic interaction becomes a kind of stable colloidal state.The example of metallic colloid thing has collaurum, collargol etc.

In some embodiments, be coated with on the pad of said test strips can with the albumen of the colloid gold label of people's antibodies.In some embodiments, be coated with the anti-human IgG antibody of colloid gold label on the pad of said test strips, the staphylococcal protein A of colloid gold label, or the streptococcal protein G of colloid gold label.

In some embodiments, also be coated with second kind of albumen of metallic colloid substance markers on the pad 2.Said second kind of albumen is different from first kind of albumen of said metallic colloid substance markers.In some embodiments, said second kind of albumen can specificity combine Quality Control albumen, but does not combine the people's antibody in the sample.Be described below, this second kind of albumen is the Quality Control tester, is used to show whether the test paper function is normal.Second kind of albumen can be selected according to actual conditions, for example, can be rabbit antibody, murine antibody, goat-anti body, pig antibody or other albumen that is fit to that does not combine people's antibody.Said second kind of albumen can form second kind of albumen of metallic colloid mark through above-mentioned method.In some embodiments, said second kind of albumen and said first kind of albumen are coated on the pad 2 with the form of potpourri.

In some embodiments; The detection film 3 of test strips 100 is provided with detection zone separated from one another 5 and Quality Control district 6; And said detection zone 5 more is close to pad 2 than said Quality Control district 6; Wherein said detection zone 5 is coated with the detection Western blot, and said Quality Control district 6 is coated with the Quality Control Western blot.The enough distances in interval make the sample of chromatography to contact with the detection Western blot of detection zone 5 and the Quality Control Western blot in Quality Control district 6 respectively, and can not disturb mutually between said detection zone 5 and the said Quality Control district 6.Detection zone 5 more is close to pad 2, makes the sample of chromatography in the chromatography process, contacts with the detection Western blot earlier, and then contacts with the Quality Control Western blot.

In some embodiments, the said detection Western blot on each said test strips is a kind of material that is selected from following group: AMA-M2 Western blot, SP100 Western blot, GP210 Western blot, SLA Western blot and LKM-1 Western blot.In some embodiments, the detection Western blot that encapsulates on the different test strips is inequality.These detect Western blot can specificity combine its corresponding antibody; Can specificity combine AMA-M2 antibody like the AMA-M2 Western blot; The SP100 Western blot can specificity combine SP100 antibody; The GP210 Western blot can specificity combine GP210 antibody, and the SLA Western blot can specificity combine SLA antibody, and the LKM-1 Western blot can specificity combine LKM-1 antibody.These albumen can carry out the biology expression and the separation and purification of albumen through the technology of routine, thereby obtain these albumen known in this field.The example of said Western blot is seen table 1.

AMA-M2 antigen, i.e. 2-oxygen acidohydrogenase complex (2-OADC) on the mitochondria, it comprises various ingredients, for example pyruvic dehydrogenase, 2-oxygen acidohydrogenase, 2-oxygen glutatate dehydrogenase, X protein etc.In some embodiments, the target antigen of AMA-M2 antibody is a kind of pyruvic dehydrogenase that connected, and the antigen protein triplet of 2-oxygen acidohydrogenase and 2-oxygen glutatate dehydrogenase (is seen; For example; Jiang Xiaohua, Zhong Renqian etc., the clonal expression of M2 autoantigen and triplet thereof and Preliminary Identification; China's digestion magazine, 2001 (21): 530-533).AMA-M2 susceptibility in PBC can reach more than 90%.

SP100 albumen is the nucleus albumen of 1 phosphorylation, and molecular mass corresponding during electrophoresis is 100,000, is a transcriptional activators.

The principal ingredient of GP210 is a nuclear membrane glycoprotein; Be that a kind of relative molecular mass is 210 in the nucleopore compound; 000 complete transmembrane protein (is seen, for example: Nakamura M etc., Increased exp ression of nuclear envelope gp210antigen in small bile ducts in primary biliary cirrhosis [J] .JAutoimmun; 2006,26 (2): 138-145).

SLA is a kind of soluble protein antigen in the cytoplasm of liver, and organizing at pancreas etc. also has expression.The molecular weight of SLA albumen is that 50KD (sees Wies I etc., Identigication of target antigen for SLA/LP autoantibodies in autoimmune hepatitis.The Lancet, 2000; 355:1510-1515).

LKM-1 (liver/kidney microsomal; LKM) be liver/kidney microsome; The target antigen composition that causes autoimmune disease is human-cytochrome II D6, and molecular weight is that 50KD (sees Duclos-Vallee J C etc.; Conformational epitopes on CYP2D6are recognized by liver/kidney microsomal antibodies.G astroenterology, 1995; 108:470-476).

In some embodiments; Said diagnostic kit comprises two said test strips; Detection Western blot on each said test strips is a kind of material that is selected from down group: AMA-M2 Western blot, SP100 Western blot, GP210 Western blot, SLA Western blot and LKM-1 Western blot, the detection Western blot that encapsulates on the different test strips is inequality.For example; Detection Western blot on said two test strips can be respectively SLA Western blot or LKM-1 Western blot, and the detection Western blot on perhaps said two test strips also can be selected from AMA-M2 Western blot, SP100 Western blot and GP210 Western blot respectively.

In some embodiments; Said diagnostic kit comprises three said test strips; Detection Western blot on each said test strips is a kind of material that is selected from down group: AMA-M2 Western blot, SP100 Western blot, GP210 Western blot, SLA Western blot and LKM-1 Western blot, the detection Western blot that encapsulates on the different test strips is inequality.For example, the detection Western blot on said three test strips can be respectively AMA-M2 Western blot, SP100 Western blot or GP210 Western blot.

In some embodiments; Said diagnostic kit comprises four said test strips; Detection Western blot on each said test strips is a kind of material that is selected from down group: AMA-M2 Western blot, SP100 Western blot, GP210 Western blot, SLA Western blot and LKM-1 Western blot, the detection Western blot that encapsulates on the different test strips is inequality.For example, the detection Western blot on said four test strips can be respectively AMA-M2 Western blot, SP100 Western blot, GP210 Western blot or SLA Western blot.For example, the detection Western blot on said four test strips also can be respectively AMA-M2 Western blot, SP100 Western blot, GP210 Western blot or LKM-1 Western blot.

Detect Western blot and can be coated on the detection zone 5 that detects film 3 with appropriate method.In some embodiments, can be dissolved in the suitable damping fluid detecting albumen, again through fix, encapsulate, adsorb, modes such as spraying, printing are coated on detection zone.Suitable damping fluid comprises, borate buffer solution, carbonate buffer solution, phosphate buffer, Tris-HCl damping fluid, Tris-phosphate buffer, acetate buffer or barbitol buffer solution etc., and its pH value can be 6.5-7.5, or 7.0-7.4.Can contact a period of time with detection zone with containing the damping fluid that detects albumen, remove the damping fluid after the contact then.Again for example, can be sprayed at or be coated on detection zone, remove solvent composition through modes such as dryings again containing the damping fluid that detects albumen.Again for example, can be imprinted on or be compressed on detection zone with detecting albumen.

In some embodiments, said Quality Control Western blot be can with the antibody or the albumen of antibodies in the sample, perhaps can with the protein bound antibody or the albumen of said colloid gold label.In some embodiments, also be coated with the albumen of second kind of colloid gold label on the pad of said test strips, and said Quality Control Western blot can combine the albumen of said second kind of colloid gold label.Said Quality Control albumen can be antibody.When said second kind of albumen was antibody, said Quality Control Western blot can also be staphylococcal protein A or streptococcal protein G.

Use and detect the similar method for coating of Western blot, can the Quality Control Western blot be coated in the Quality Control district 6 of detection film 3.Whether said Quality Control Western blot can be used for monitoring the chromatography effect normally takes place.If the Quality Control Western blot can detect the albumen of metal marker substance markers, then show sample from sample pad 1 process pad 2, and taken away the albumen of the metallic colloid substance markers that encapsulates on the pad 2, continue to move to detection film 3 then.This this testing result of explanation is effective.If the Quality Control Western blot does not detect any signal, then point out test strips possibly not have operate as normal, testing result is invalid.

The Quality Control Western blot that encapsulates in detection Western blot that encapsulates on the detection zone 5 and the Quality Control district 6 can be any suitable shapes.In some embodiments, detection Western blot and/or Quality Control Western blot is shaped as mottled or ribbon.In some embodiments, detecting Western blot and Quality Control Western blot all is mottled or all is ribbon.Here said mottled can be regular shapes or irregularly shaped such as circle, ellipse.

In some embodiments; Said sample pad is to be processed by spun glass, polyester film, cellulose filter paper or nonwoven fabrics; Said pad is to be processed by spun glass, polyester film, cellulose filter paper or nonwoven fabrics; With said detection film be to process by nitrocellulose filter, and said supporting baseplate by PVC plate or other hard ground and the material that does not absorb water process.In some embodiments, the adsorptive pads 4 of test strips 100 can be the tablet of being processed by absorption paper.Spun glass, polyester film, cellulose filter paper, nonwoven fabrics, absorption paper and nitrocellulose filter all are known materials, and those skilled in the art can obtain through commercial sources at an easy rate.The model of these materials and specification can be selected according to actual needs.The model of the spun glass that for example, is suitable for has Ahlstrom8964, Ahlstrom8975, GF-06 and GF-08.The model of the nitrocellulose filter that is suitable for can comprise Millipore135, Sartorius CN140 and Vivid 170 etc.

In some embodiments, each test strips also comprises a supporting baseplate 7.In some embodiments, supporting baseplate 7 can be dull and stereotyped, as shown in Figure 2.Other parts of test strips 100 can be fixed on the flat supporting baseplate 7 through the mode of pasting, and for example use adhesive tape to stick on the supporting baseplate.Said adhesive tape can be adhesive tape well known in the art or translucent adhesive tape.Again for example, also can on supporting baseplate, apply sticking material, other parts with test strips 100 stick on the supporting baseplate again.

In some embodiments, kit further comprises at least one firm banking, is fixed with at least one said test strips on said at least one firm banking.Base can be processed by suitable materials such as plastics.In some embodiments, shown in Fig. 3 (a) and 3 (b), the plate face edge of base 8 can be provided with continuous projection 9, and projection 9 defines the accommodation section 20 that can hold single test strips on base 8.Although the illustrated accommodation section of being defined by continuous projection 9 20 is the for example closed figures of rectangle; But it will be appreciated by those skilled in the art that; Can also on base 8, form the projection of suitable shapes such as discontinuous point-like, strip, and. by the accommodation section that projection defined also can be the non-closed figures such as "=", " C " shape.For example, can two row spot-like projections be set respectively in two opposite edges of base 8, this two row spot-like projections defines the groove shape accommodation section that can hold single test strips on base.Further, the shape of accommodation section and/or size can be suitable with test strips.

In some embodiments, kit also can comprise can be 8 that cooperate with firm banking, with firm banking 8 the same number of loam cakes 10.As shown in Figure 4, each loam cake 10 is equipped with application of sample mouth separated from one another 11 and watch window 12.Loam cake 10 can match with the firm banking that is provided with test strips 8 through stickup or manner.When loam cake 10 cooperated with firm banking 8, the application of sample mouth 11 of loam cake 10 was positioned at sample pad 1 top of test strips, and watch window 12 is positioned at detection film 3 tops on the test strips, preferably exposed the detection zone 5 and Quality Control district 6 that detect film 3.Fig. 5 (a) and 5 (b) have schematically shown loam cake 10 that is fitted to each other and the structure that is provided with the firm banking 8 of single test strips.

In some embodiments, kit further comprises at least one firm banking, is fixed with said at least two test strips on said at least one firm banking.For example, said kit comprises a firm banking 8 ', and it is provided with at least two test strips 100.In some embodiments, be fixed with two or five said test strips on said at least one firm banking, and said test strips is separate.In some embodiments, said test strips can be separated from each other.

In some embodiments, the plate face of said firm banking is provided with projection, and wherein said test strips is placed in the accommodation section of being defined by said projection, can hold single test strips.For example, on the plate face of firm banking 8 ' continuous projection 9 ' can be set, projection 9 ' defines at least two accommodation sections 20 that can hold single test strips 100 respectively, makes that said at least two test strips 100 are separated from one another.Again for example, on the plate face of firm banking 8 ' continuous projection 9 ' can be set, projection 9 ' defines two to five accommodation sections 20 that can hold single test strips 100 respectively, makes that said two to five test strips 100 are separated from one another.Can test strips 100 be fixed in the accommodation section separately with the mode of pasting.Fig. 6 (a) and Fig. 6 (b) show the vertical view and the cut-open view of the firm banking 8 ' that is provided with continuous projection 9 ' that has two accommodation sections respectively, and Fig. 7 (a) and Fig. 7 (b) show the vertical view and the cut-open view of the firm banking 8 ' that is provided with continuous projection 9 ' that has three accommodation sections respectively.Likewise, those skilled in the art can know the structure of the firm banking that has four, five or more accommodation sections at an easy rate according to the exemplary plot of two and three accommodation sections.Certainly, the projection 9 ' on the firm banking 8 ' also can be configured to the projection of a plurality of point-like, strip as top introduction, and the shape of each accommodation section that projection 9 ' defined also can be non-closed figure (a for example groove shape).The shape and/or the size of preferred each accommodation section are suitable with test strips 100.In some embodiments, the plate face of said firm banking is provided with projection, and said projection defines at least two accommodation sections that can hold single test strips.

In some embodiments; Kit also comprise at least one can with the matching used loam cake of said firm banking; Each said loam cake has application of sample mouth and watch window; Said application of sample mouth is positioned at the said sample pad top of said test strips, and said watch window is positioned at the said detection film top on the said test strips.For example, kit also can comprise a loam cake 10 ' that can cooperate with firm banking 8 ', and said loam cake 10 ' is provided with at least two application of sample mouths 11 ' separated from one another, and the mode that said loam cake 10 ' can be pasted or fasten cooperates with firm banking 8 '.When loam cake 10 ' covered with the firm banking 8 ' that is provided with two test strips 100 at least, each application of sample mouth 11 ' laid respectively at sample pad 1 top of corresponding test strips 100.In some embodiments, said loam cake 10 ' also can comprise at least two watch windows 12 ' separated from one another.When loam cake 10 ' covered with the firm banking 8 ' that is provided with two test strips 100 at least, each watch window 12 ' laid respectively at detection film 3 tops of corresponding test strips, preferably exposed the detection zone 5 and Quality Control district 6 of this detection film 3.Fig. 6 (c) shows the vertical view of the loam cake that has two application of sample mouths and two watch windows; Fig. 7 (c) shows the vertical view of the loam cake that has three application of sample mouths and three watch windows; Likewise, those skilled in the art can know the superstructure that has four, five or more add appearance mouth and watch window at an easy rate according to Fig. 6 (c) and Fig. 7 (c).It will be appreciated by those skilled in the art that; Said at least two watch windows 12 ' separated from one another and 12 " also can substitute with a coherent watch window; at loam cake 10 ' and the firm banking 8 ' and 8 that is provided with at least two or three test strips 100 " when covering; This watch window is positioned at detection film 3 tops of all test strips, preferably exposes all detection zones that detect film 35 and Quality Control district 6.

In some embodiments, said diagnostic kit comprises two or more said firm bankings, wherein is fixed with at least one said test strips respectively on each said firm banking.In some embodiments, said two or more said firm banking is separate.For example, said diagnostic kit can comprise two separate firm bankings, and wherein each all is fixed with at least one test strips.Again for example, said diagnostic kit can comprise two separate firm bankings, and one of them is fixed with at least two test strips, and another is fixed with at least one test strips.Again for example, said diagnostic kit can comprise three separate firm bankings, and wherein two bases are fixed with two test strips respectively, and another is fixed with a test strips.For example, said diagnostic kit can comprise five separate firm bankings, and wherein each base is fixed with a test strips respectively.In some embodiments; Said diagnostic kit also include can with the matching used loam cake of said firm banking; Loam cake has application of sample mouth and watch window; Said application of sample mouth is positioned at the said sample pad top of said test strips, and said watch window is positioned at the said detection film top on the said test strips.

In some embodiments, said at least two test strips in the said diagnostic kit are parallel to each other.In some embodiments; The detection zone 5 and/or the Quality Control district 6 that are arranged at least two test strips 100 on the firm banking 8 ' are arranged on the same straight line basically; Promptly the detection zone 5 of at least two test strips 100 is arranged point-blank basically, and/or Quality Control district 6 is arranged on same the straight line basically.In some embodiments, at least two test strips 100 are arranged in parallel on firm banking 8 '.

In some embodiments, diagnostic kit comprises that further one or more are selected from down the ingredient of group: firm banking, loam cake, drying agent, dropper, instructions, color label, sample collection tube and sample diluting liquid.Said dropper can be used for the imbitition sample, and can be used for fluid sample is added drop-wise to sample pad 1.Said drying agent can keep the humidity of kit in suitable scope.On the said instructions with information such as method of application, store method, product batch numbers.

In some embodiments, said kit can also comprise the polybag of at least one lucifuge, sealing, and said kit wherein is housed.

In some embodiments, said kit can be used in and detects AMA-M2 antibody, SP100 antibody, GP210 antibody, SLA antibody or LKM-1 antibody.For example, said kit can be used for whether existing in the test sample one or more above-mentioned antibody.If there are one or more above-mentioned antibody in the sample, it can be caught by the detection albumen of the correspondence on the film to be detected in sample chromatography process, thereby is detected at detection zone.

For example, add sample after, react on the relevant position that can see detection zone 5 and Quality Control district 6 in 3～5 minutes and dark band 13 all occurs; 14; Shown in Fig. 8 (a) and 8 (b), this expression testing result is positive, contains the antibody that can combine with the detection protein-specific in the interpret sample.For another example, behind the application of sample, only 6 a dark band 14 occurs in the Quality Control district, dark band 13 (shown in Fig. 8 (c)) does not appear in detection zone 5, and this expression testing result is negative, do not contain in the interpret sample can with detect the antibody that protein-specific combine.For another example, behind the application of sample, dark band 14 does not appear in Quality Control district 6, and this moment, no matter whether detection zone 6 had band (shown in Fig. 8 (d), 8 (e)) to occur, explained that all test paper lost efficacy, and testing result is invalid.

In some embodiments, the diagnostic kit that provides of the application can be used for the assisted diagnosis disease.In some embodiments, said diagnostic kit can be used for the assisted diagnosis autoimmune liver disease, for example, and oneself immunity hepatitis (AIH) and/or PBC (PBC).

LKM-1 antibody and SLA antibody are respectively the significant antibody of II type and III type AIH, and its detection specificity is high.Has the meaning of making a definite diagnosis.Through whether having LKM-1-1 antibody and/or SLA antibody in the test sample, can auxiliary diagnosis sample source person whether suffer from AIH.

Three kinds of serological index of PBC diagnosis are AMA-M2 antibody, SP100 antibody and GP210 antibody.Wherein AMA-M2 antibody is main serotype index, in most of PBC patients, can detect.SP100 antibody can detect in 31% patient PBC, but in other autoimmune liver diseases, detect less than.In addition, SP100 antibody is also found in patient PBC of 48%AMA-M2 negative antibody.Anti-nuclear membrane glycoprotein GP210 antibody can detect in 10% patient PBC; Be the high degree of specificity antibody of PBC; Its specificity seldom comes across among oneself immunity hepatitis, rheumatoid arthritis, Sjogren syndrome and the polymyositis patient up to more than 96%.Detecting GP210 antibody has great importance for AMA-M2 negative antibody or PBC patient and its laboratory, pathological examination, unusual the making a definite diagnosis of PBC patient that lack general clinical features.Therefore, M2, three kinds of antibody of SP100 and GP210 are measured simultaneously, and the diagnosis that improves PBC is had very big meaning.

The kit that the application provides can carry out joint inspection simultaneously to the multinomial serological index of AIH and/or PBC, helps simultaneously examination AIH and/or PBC, and the making a definite diagnosis of auxiliary AIH or PBC, to avoid mistaken diagnosis and to fail to pinpoint a disease in diagnosis.

In some embodiments, have in LKM-1 antibody and the SLA antibody one or both in the sample if detect, the person possibly suffer from AIH then to point out the sample source.

In some embodiments, have in AMA-M2 antibody, SP100 antibody and the GP210 antibody one or more in the sample if detect, the person possibly suffer from PBC then to point out the sample source.

In some embodiments, do not have in LKM-1 antibody, SLA antibody, AMA-M2 antibody, SP100 antibody and the GP210 antibody any in the sample if detect, the person possibly neither suffer from AIH then to point out the sample source, does not also suffer from PBC.

The diagnostic kit that please provide in this uses colloidal gold immunity chromatography.This method is to use colloidal gold-labeled method; With collaurum as tracer; With the fibre strip chromatographic material is solid phase; Make sample solution swimming on chromatography strip through capillary effect; Make that immune response takes place the acceptor (like antigen or antibody) to determinand on determinand and the pad in the sample, and immune response takes place and be trapped, and then form macroscopic aubergine band with antigen (or antibody) on the fibre strip chromatographic material; Obtain experimental result intuitively, reach the purpose (Sikowicz G et al.One-step chromatographic immunoassay for qualitative determination of choricogonadotropin in urine.Clin Chem.1990 (36) 1579-1586.) of fast detecting.Only need during use sample pipetting volume on sample pad, just situation to occur and judge the yin and yang attribute result in several minutes according to aubergine band on the detection line.Advantages such as compare with other detection methods, detection time is short, only needs 5～10 minutes, and operating personnel need not training, do not receive place personnel's limit, and are easy and simple to handle, quick, need not cryopreservation, and accumulating is convenient.

This test paper has high specific, high sensitivity, pin-point accuracy in addition, can satisfy clinical rapid screening PBC and AIH, for patient's diagnoses and treatment early provides condition, simultaneously, also can satisfy the demand of basic unit laboratory, instant detection, bedside detection.Therefore, the diagnostic kit that provides of the application is compared with existing product and is had remarkable advantages.

Example 1:

The structure of kit

A diagnostic kit is provided in this example, and it comprises two test strips as shown in Figure 1, and wherein each test strips 100 all comprises sample pad 1, pad 2, detection film 3 and the adsorptive pads 4 of overlap joint successively, and supporting baseplate 7.All be coated with anti-people's antibody of colloid gold label on the pad 2 of each test strips 100, and the rabbit igg of colloid gold label.All be provided with detection zone separated from one another 5 and Quality Control district 6 on the detection film 3 of each test strips 100, and said detection zone 5 more is close to pad 2 than said Quality Control district 6.Detection zone 5 is coated with the detection Western blot, and Quality Control district 6 is coated with the Quality Control Western blot.Detection Western blot on two test strips 100 is different, is respectively: soluble liver antigen SLA and LKM-1.The Quality Control Western blot of each test strips 100 is identical, all is goat anti-rabbit igg.Said diagnostic kit also comprises a firm banking as shown in Figure 68 ' and loam cake 10 '.Projection 9 ' on the firm banking 8 ' defines two accommodation sections 20 that can hold and separate test strips 100; Two test strips 100 are placed on respectively in the accommodation section 20 with being arranged in parallel; And on same straight line, Quality Control district 6 is also basically on same straight line basically for the detection zone 5 on the detection film 3.Loam cake 10 ' is fastened on after firm banking 8 ' goes up, and the application of sample mouth 11 ' on the loam cake 10 ' is respectively over against the sample pad 1 of each test strips 100, and detection window 12 ' is respectively over against the detection film 3 of each test strips 100.Said diagnostic kit also can contain not shown dropper, drying agent, instructions, color label, sample collection tube and sample diluting liquid.

The method that detects with kit

Carry out human sample's detection with the kit in the above-mentioned example.Collect to come from 100 parts of clinical serum that are diagnosed as oneself immunity hepatitis (AIH) patient.For each patient's sample, draw an amount of blood sample with dropper, be added drop-wise to two application of sample mouths respectively.After leaving standstill 5 minutes under the room temperature condition, observe and the record result.

Testing result to 100 parts of AIH patient diagnoseds' serum is carried out statistics and analysis, to verify sensitivity and the specificity of this kit to the AIH diagnosis.Two joint inspection indexs of statistics AIH: SLA antibody and LKM-1 detection of antibodies result, see table 2.Calculating is for the diagnosis sensitivity of each joint inspection index: diagnosis sensitivity (%)=positive findings sample number/specimen sum * 100%.The result shows that this kit is respectively 31% and 39% to the diagnostic sensitivity of SLA antibody and LKM-1 antibody.In 100 seized duplicate samples, as long as arbitrary test positive in two joint inspection indexs of AIH, then this sample is judged as AIH patient's sample.The result shows there are 62 parts in 100 duplicate samples and are detected the AIH positive findings, shows that this kit can reach 62% to AIH patient's diagnosis.In addition, use these AIH patients' serum to detect PBC joint inspection index (being AMA-M2 antibody, SP100 antibody and GP210 antibody) separately, the result that nearly all is negative explains that this kit has higher specificity to the diagnosis of AIH.

Table 2

Example 2:

The structure of kit

A diagnostic kit is provided in this example, and it comprises three test strips as shown in Figure 1, and wherein each test strips 100 all comprises sample pad 1, pad 2, detection film 3 and the adsorptive pads 4 of overlap joint successively.All be coated with anti-people's antibody of colloid gold label on the pad 2 of each test strips 100, and the rabbit igg of colloid gold label.All be provided with detection zone separated from one another 5 and Quality Control district 6 on the detection film 3 of each test strips 100, and said detection zone 5 more is close to pad 2 than said Quality Control district 6.Detection zone 5 is coated with the detection Western blot, and Quality Control district 6 is coated with the Quality Control Western blot.Detection Western blot on three test strips 100 is different, is respectively: AMA-M2, SP100 and GP210 albumen.The Quality Control Western blot of each test strips 100 is identical, all is goat anti-rabbit igg.Said diagnostic kit also comprises a firm banking as shown in Figure 78 " and loam cake 10 ".Firm banking 8 " on projection 9 " define three accommodation sections 20 that can hold and separate test strips 100; Three test strips 100 are placed on respectively in the accommodation section 20 with being arranged in parallel; And on same straight line, Quality Control district 6 is also basically on same straight line basically for the detection zone 5 on the detection film 3.Loam cake 10 " be fastened on firm banking 8 " go up after, loam cake 10 " on application of sample mouth 11 " respectively over against the sample pad 1 of each test strips 100, detection window 12 " respectively over against the detection film 3 of each test strips 100.Said diagnostic kit also can contain not shown dropper, drying agent, instructions, color label, sample collection tube and sample diluting liquid.

The method that detects with kit

Carry out human sample's detection with the kit in the above-mentioned example.Collect to come from 100 parts of clinical PBC patients diagnosed serum.For each patient's sample, draw an amount of blood sample with dropper, be added drop-wise to three application of sample mouths respectively.After leaving standstill 5 minutes under the room temperature condition, observe and the record result.

Testing result to 100 parts of PBC patient diagnoseds' serum is carried out statistics and analysis, to verify sensitivity and the specificity of this kit to the PBC diagnosis.Three joint inspection indexs of statistics PBC: AMA-M2 antibody, SP100 antibody and GP210 detection of antibodies result, see table 3.Calculating is for the diagnosis sensitivity of each joint inspection index: diagnosis sensitivity (%)=positive findings sample number/specimen sum * 100%.The result shows that this kit is respectively 95%, 32% and 35% to the diagnostic sensitivity of AMA-M2 antibody, SP100 antibody, GP210 antibody.In 100 seized duplicate samples, as long as arbitrary test positive in three joint inspection indexs of PBC, then this sample is judged as PBC patient's sample.The result shows there are 99 parts in 100 duplicate samples and are detected the PBC positive findings, shows that this kit can reach 99% to PBC patient's diagnosis.In addition, use these PBC patients' serum to detect AIH joint inspection index (being SLA antibody and LKM-1 antibody) separately, the result that nearly all is negative explains that this kit has higher specificity to the diagnosis of PBC.

Table 3

Example 3:

Specificity is analyzed:

From clinical, collect healthy blood donor's serum and each 300 parts of contrast disease patient (non-PBC, patient AIH comprise other autoimmune diseases such as rheumatoid arthritis) serum, above-mentioned serum is detected with the kit in example 1 and 2.

Statistics detects feminine gender and the positive findings that obtains with this kit, and calculates the specificity for each seized antibody according to formula: specificity (%)=negative findings sample number/specimen sum * 100%.The result shows; The kit of example 1 is respectively the specificity of SLA antibody and LKM-1 antibody: 98.8% and 99.0%, and the kit in the example 2 is respectively 99.5%, 99.2% and 99.0% to the specificity of AMA-M2 antibody, SP100 antibody and GP210 antibody.The result shows, example 1 is all very high to the specificities of two and three detection indexs of correspondence with 2 kit.

Claims

1. a diagnostic kit that is used for the diagnosis of autoimmune hepatopathy comprises at least two test strips, and wherein said test strips comprises sample pad (1), pad (2), detection film (3) and the adsorptive pads (4) of overlap joint successively, and supporting baseplate (7), wherein:

Be coated with on the pad of said test strips (2) can with the albumen of the metallic colloid mark of people's antibodies;

Be coated with a kind of detection Western blot on the detection zone (5) of the detection film (3) of said test strips;

Also be coated with a special quality control Western blot in the Quality Control district (6) of the detection film (3) of said test strips; And

Said detection Western blot and said Quality Control Western blot on the detection film (3) of said test strips are separated from one another, and said detection Western blot more is close to pad (2) than said Quality Control Western blot.

2. diagnostic kit as claimed in claim 1, wherein said sample pad (1) are to be processed by spun glass, polyester film, cellulose filter paper or nonwoven fabrics; Said pad (2) is to be processed by spun glass, polyester film, cellulose filter paper or nonwoven fabrics;

Said detection film (3) is to be processed by nitrocellulose filter; And said supporting baseplate (7) is processed by the PVC plate.

3. diagnostic kit as claimed in claim 1 is characterized in that the shape of said detection Western blot and Quality Control Western blot is respectively mottled or ribbon.

4. diagnostic kit as claimed in claim 1; Wherein the detection Western blot on each said test strips is a kind of material that is selected from down group: AMA-M2 Western blot, SP100 Western blot, GP210 Western blot, SLA Western blot and LKM-1 Western blot, and the detection Western blot that encapsulates on the different test strips is inequality.

5. diagnostic kit as claimed in claim 1; It is characterized in that comprising two said test strips; Detection Western blot on each said test strips is a kind of material that is selected from down group: AMA-M2 Western blot, SP100 Western blot, GP210 Western blot, SLA Western blot and LKM-1 Western blot, the detection Western blot that encapsulates on the different test strips is inequality.

6. diagnostic kit as claimed in claim 1; It is characterized in that comprising three said test strips; Detection Western blot on each said test strips is a kind of material that is selected from down group: AMA-M2 Western blot, SP100 Western blot, GP210 Western blot, SLA Western blot and LKM-1 Western blot, the detection Western blot that encapsulates on the different test strips is inequality.

7. diagnostic kit as claimed in claim 1; It is characterized in that comprising four said test strips; Detection Western blot on each said test strips is a kind of material that is selected from down group: AMA-M2 Western blot, SP100 Western blot, GP210 Western blot, SLA Western blot and LKM-1 Western blot, the detection Western blot that encapsulates on the different test strips is inequality.

8. diagnostic kit as claimed in claim 1; It is characterized in that comprising five said test strips; Detection Western blot on each said test strips is a kind of material that is selected from down group: AMA-M2 Western blot, SP100 Western blot, GP210 Western blot, SLA Western blot and LKM-1 Western blot, the detection Western blot that encapsulates on the different test strips is inequality.

9. diagnostic kit as claimed in claim 1, wherein the Quality Control Western blot be with sample in the antibody or the albumen of antibodies, perhaps with the protein bound antibody or the albumen of said colloid gold label.

10. diagnostic kit as claimed in claim 1 also be coated with the albumen of second kind of colloid gold label on the pad of wherein said test strips (2), and said Quality Control Western blot can combine the albumen of said second kind of colloid gold label.

11. diagnostic kit as claimed in claim 1 is characterized in that further comprising at least one firm banking, is fixed with said at least two test strips on said at least one firm banking.

12. diagnostic kit as claimed in claim 11 be fixed with two to five said test strips on wherein said at least one firm banking, and said test strips is separate.

13. like claim 11 or 12 described diagnostic kits, it is characterized in that the plate face of said firm banking is provided with projection, wherein said test strips is placed in the accommodation section of being defined by said projection, can hold single test strips.

14. like claim 11 or 12 described diagnostic kits, it is characterized in that the plate face of said firm banking is provided with projection, said projection defines at least two accommodation sections that can hold single test strips.

15. like claim 11 or 12 described diagnostic kits, it is characterized in that comprising two or more said firm bankings, wherein be fixed with at least one said test strips respectively on each said firm banking.

16. like claim 11 or 12 described diagnostic kits; It is characterized in that also comprising and said firm banking and the matching used loam cake of said test strips; Each said loam cake has application of sample mouth and watch window; Said application of sample mouth is positioned at the said sample pad top of said test strips, and said watch window is positioned at the said detection film top on the said test strips.

17. diagnostic kit as claimed in claim 1, wherein said at least two test strips are parallel to each other.

18. diagnostic kit as claimed in claim 17; The detection Western blot that encapsulates on the detection film (3) of wherein said test strips is positioned on the same horizontal line basically, and the Quality Control Western blot that encapsulates on the detection film (3) of said test strips is positioned on the same horizontal line basically.

19. diagnostic kit as claimed in claim 1 is characterized in that further comprising that one or more are selected from down the ingredient of group: the firm banking, loam cake, drying agent, dropper, instructions, color label, sample collection tube and the sample diluting liquid that are used for fixing said test strips.