A kind of colloidal gold strip that detects anti-cyclic citrulline peptide antibody and rheumatoid factor
technical field
The utility model belongs to Lateral Flow Strip field, is specifically related to a kind of colloidal gold strip that detects anti-cyclic citrulline peptide antibody and rheumatoid factor.
background technology
Rheumatoid arthritis (Rheumatoid arthritis abridge RA.) is rheumatismal a kind of, is mainly to cause because carry out for a long time contact or the wind of cold water.Rheumatoid disease activity generally usually continues 3-6 month, and wandering pain is body arthralgia, and migration is indefinite, time and at shoulder, time and at elbow, time and at upper limbs.It is a kind of general autoimmunity disease that chronic aggressivity arthritis is feature of take.The lesion characteristic of rheumatoid arthritis is synovitis, and the articular cartilage causing thus and destruction of bone, finally causes joint deformity.Synovitis is outbreak repeatedly lastingly, the destruction that can cause cartilage and bone in joint, and joint function disturbance, if without regular treatment, approximately 75% patient there will be maimed person in 3 years.
Rheumatoid arthritis is distributed in all over the world, and the morbidity rate in different crowd is 0.18% ~ 1.07%, and its morbidity has certain racial difference, and Indians are higher than white people, and white people are higher than Asia yellow, at total number of patients of China, surpasses 5,000,000.Rheumatoid arthritis all can be fallen ill in each age, and the peak age, general women's morbidity was more than the male sex about 30 ~ 50 years old.
Up to now, main still according to clinical manifestation for the early diagnosis of RA; And serology means are mainly to detect rheumatoid factor (RF).RF can be used as the sensitive serologic marker thing of RA, and specificity is approximately 70%.1998, external, the anti-cyclic citrulline peptide antibody of reported first (anti-cyclic cirullinated peptide antibodies, anti-CCP antibody) detection, RA being had to higher specificity, can make up the defect of the poor specificity of RF, is the blood serum designated object that RA is new.Domestic The People's Hospital of Peking University in 2012 take the lead criteria for classification that national 12Jia hospital participates in having proposed early stage rheumatoid arthritis (ERA) jointly wherein Serological testing have one: anti-citrulline protein antibodies (comprising anti-CCP antibody and anti-MCV antibody), has at least a high titre positive.
Therefore, anti-cyclic citrullinated peptide (CCP) antibody and rheumatoid factor (RF) joint-detection diagnostics classes rheumatic arthritis promptly and accurately.Therefore, diagnosis morning for rheumatoid arthritis has very important significance.
Anti-cyclic citrullinated peptide (CCP) antibody and rheumatoid factor (RF) detection kit (colloidal gold method) adopt antigen-antibody reaction and the immunochromatographiassays assays technology of high degree of specificity.
Anti-cyclic citrullinated peptide (CCP) TPPA adopts dual-antigen sandwich method immunoassays principle: in test strips, contain and be coated in advance anti-cyclic citrullinated peptide (CCP) antigen of colloid gold label on glass fibre membrane and be coated on anti-cyclic citrullinated peptide (CCP) antigen on nitrocellulose filter.During detection, if sample contains anti-cyclic citrullinated peptide (CCP) antibody, can in anti-cyclic citrullinated peptide (CCP) antibody test district, form double antigens sandwich compound, have mauve band; If negative sample, does not have aubergine band in anti-cyclic citrullinated peptide (CCP) antibody test district.
Rheumatoid factor (RF) is measured and is adopted dual-antigen sandwich method immunoassays principle: in test strips, contain the coated in advance hot polymerization human IgG (as antigen) on glass fibre membrane and be coated on the hot polymerization human IgG (as antigen) on nitrocellulose filter.During detection, if sample is positive, can on rheumatoid factor (RF) test section, form double antigens sandwich compound, have mauve band; If negative sample, does not have aubergine band on rheumatoid factor (RF) test section.
summary of the invention
The purpose of this utility model is to provide a kind of colloidal gold strip that simultaneously detects anti-cyclic citrulline peptide antibody and rheumatoid factor.
The utility model is by the following technical solutions:
A kind of colloidal gold strip that detects anti-cyclic citrulline peptide antibody and rheumatoid factor, comprise base plate, on base plate, be attached with tight connected sample pad from left to right, glass fibre membrane, nitrocellulose filter and absorbing membrane and absorbent filter, when print pad is connected with glass fibre film close, sample pad is being pressed glass fibre membrane 1-3mm, when glass fibre membrane is connected with cellulose nitrate film close, glass fibre mould nitrocellulose filter 1-3mm, the nitrocellulose filter mould that absorbs water while being closely connected with absorbing membrane nitrocellulose filter 1-3mm, when absorbing membrane is closely connected with absorbent filter, absorbent filter is being pressed absorbing membrane 1-3mm.
On described glass fibre element film, absorption has cyclic citrullinated peptide antigen and the hot polymerization human IgG of colloid gold label.
On described nitrocellulose filter, be coated with cyclic citrullinated peptide antigen, hot polymerization human IgG (as antigen) and mouse-anti human IgG.
Cyclic citrullinated peptide antigen, hot polymerization human IgG and mouse-anti human IgG are all ribbon and are coated on nitrocellulose filter successively from left to right.
Absorbent filter can absorb the liquid that is not less than 500 microlitres.
The utility model adds to tracer liquid in sample pad while using, sample pad can fully absorb tracer liquid, the first chromatography of tracer liquid contacts abundant reaction completely with the gold mark antigen of spraying to glass fibre membrane, chromatography reacts to nitrocellulose filter again, when anti-cyclic citrullinated peptide (CCP) antibody and rheumatoid factor (RF), the content in sample, has mauve band in corresponding test section higher than test strips to its lowest detection in limited time.Anti-cyclic citrullinated peptide (CCP) antibody and the content of rheumatoid factor (RF) in sample, do not have mauve band in corresponding test section lower than test strips to its lowest detection in limited time.
Negative findings: only have purple line to occur at the coated place of mouse-anti human IgG of nitrocellulose filter, as shown in Figure 2 b.
Positive findings: there is purple line at coated place except the mouse-anti human IgG at nitrocellulose filter, at the coated place of cyclic citrullinated peptide antigen, hot polymerization human IgG of nitrocellulose filter, also there are one or two purple lines appearance, specifically: at the coated place of mouse-anti human IgG of nitrocellulose filter, the antigen coated place of cyclic citrullinated peptide of nitrocellulose filter, all have purple line to occur (seeing Fig. 2 d), show cyclic citrullinated peptide antibody positive; At the coated place of mouse-anti human IgG of nitrocellulose filter,, the coated place of hot polymerization human IgG of nitrocellulose filter all has purple line to occur (seeing Fig. 2 c), show that rheumatoid factor is positive; The coated place of mouse-anti human IgG, the cyclic citrullinated peptide antigen of nitrocellulose filter, the coated place of hot polymerization human IgG at nitrocellulose filter all have purple line to occur (seeing Fig. 2 e), illustrate that cyclic citrulline peptide antibody, rheumatoid factor are all positive.
Invalid: when nature controlling line (the coated place of mouse-anti human IgG of nitrocellulose filter) does not demonstrate red stripes, no matter whether detection line demonstrates red stripes, and it is invalid that this test card is judged to, as shown in Figure 2 a.
Simplicity of design of the present utility model, can reduce error effectively, and easy and simple to handle fast result shows image, directly perceived accurately, cost is low, highly sensitive.
accompanying drawing explanation
Fig. 1 is structural representation of the present utility model;
Fig. 2 is colour developing schematic diagram of the present utility model, 2a is that test card is judged to schematic diagram when invalid, the negative result schematic diagram of 2b, 2c is that rheumatoid factor positive findings schematic diagram, 2d are that cyclic citrullinated peptide antibody positive result schematic diagram, 2e are that cyclic citrulline peptide antibody, rheumatoid factor are positive findings schematic diagram.
embodiment
The anti-cyclic citrulline peptide antibody of detection as shown in Figure 1 and the colloidal gold strip of rheumatoid factor, comprise base plate 8, on base plate 8, be attached with tight connected sample pad 1 from left to right, glass fibre membrane 2, nitrocellulose filter 3, absorbing membrane 7 and absorbent filter 9, when print pad 1 is closely connected with glass fibre membrane 2, sample pad 1 is being pressed glass fibre membrane 2 1-3mm, when glass fibre membrane 2 is closely connected with nitrocellulose filter 3, glass fibre membrane 2 is being pressed nitrocellulose filter 3 1-3mm, when nitrocellulose filter 3 is closely connected with absorbing membrane 7, absorbing membrane 7 is being pressed nitrocellulose filter 3 1-3mm, when absorbing membrane 7 is closely connected with absorbent filter 9, absorbent filter 9 is being pressed absorbing membrane 7 1-3mm, on glass fibre element film 2, absorption has cyclic citrullinated peptide antigen and the hot polymerization human IgG of colloid gold label.; On nitrocellulose filter 3, be coated with and be all from left to right banded cyclic citrullinated peptide antigen 4, hot polymerization human IgG (as antigen) 5 and mouse-anti human IgG 6.
During use, tracer liquid is added in sample pad 1, sample pad 1 can fully absorb tracer liquid, and the first chromatography of tracer liquid contacts abundant reaction completely with the gold mark antigen of spraying to glass fibre membrane 2, then chromatography reacts to nitrocellulose filter 3.Concrete result judgement is as follows:
Negative findings: only the mouse-anti human IgG at control zone C(nitrocellulose filter 3 is coated with place) there is purple line to occur, as shown in Figure 2 b.
Positive findings: except (the coated place of mouse-anti human IgG 6 of nitrocellulose filter 3) has purple line in control C district, at surveyed area (the cyclic citrullinated peptide antigen 4 of nitrocellulose filter 3, the coated place of hot polymerization human IgG 5), also there are one or two purple lines appearance, specifically: at the coated place of mouse-anti human IgG 6 of nitrocellulose filter 3,, the coated place of cyclic citrullinated peptide antigen 4 of nitrocellulose filter 3 all has purple line to occur (seeing Fig. 2 d), show cyclic citrullinated peptide antibody positive; At the coated place of mouse-anti human IgG 6 of nitrocellulose filter 3,, the coated place of hot polymerization human IgG 5 of nitrocellulose filter 3 all has purple line to occur (seeing Fig. 2 c), show that rheumatoid factor is positive; At the coated place of mouse-anti human IgG 6 of nitrocellulose filter 3,, the cyclic citrullinated peptide antigen 4 of nitrocellulose filter 3, the coated place of hot polymerization human IgG 5 all have purple line to occur (seeing Fig. 2 e), illustrate that cyclic citrulline peptide antibody, rheumatoid factor are all positive.
Invalid: when nature controlling line (the coated place of mouse-anti human IgG 6 of nitrocellulose filter 3) does not demonstrate red stripes, no matter whether detection line demonstrates red stripes, and it is invalid that this test card is judged to, as shown in Figure 2 a.