CN105974111B - A kind of method of rheumatoid factor (IgM type) and other antigen-specific IgM antibody joint-detections - Google Patents
A kind of method of rheumatoid factor (IgM type) and other antigen-specific IgM antibody joint-detections Download PDFInfo
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- CN105974111B CN105974111B CN201610576315.XA CN201610576315A CN105974111B CN 105974111 B CN105974111 B CN 105974111B CN 201610576315 A CN201610576315 A CN 201610576315A CN 105974111 B CN105974111 B CN 105974111B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6854—Immunoglobulins
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/10—Musculoskeletal or connective tissue disorders
- G01N2800/101—Diffuse connective tissue disease, e.g. Sjögren, Wegener's granulomatosis
- G01N2800/102—Arthritis; Rheumatoid arthritis, i.e. inflammation of peripheral joints
Abstract
The present invention provides a kind of rheumatoid factor (IgM type) and other antigen-specific IgM antibody combined detection test papers, the test strips include the sample pad successively attached on polyvinyl chloride plastic sheet, conjugate pad, nitrocellulose filter and water absorption pad, are successively coated with the first detection line, adsorption line, one or more detection line and nature controlling line respectively on the nitrocellulose filter；First detection line is denaturation rabbit/human IgG, the adsorption line is the antibody of anti-human igg, the nature controlling line behaviour IgM, one or more detection line are respectively selected from one or more of specific antigen: mycoplasma pneumoniae specific antigen, influenza Type B specific antigen, toxoplasmosis pathogen-specific antigen, autoimmune antigen and anaphylactogen.The present invention also provides the kit including the test strips, the preparation method of the test strips, the applications of the method detected using the test strips or kit and the test strips or kit.
Description
Technical field
The present invention relates to a kind of test strips.Specifically, the present invention relates to a kind of rheumatoid factor (IgM type) and other are anti-
Former specific IgM antibodies combined detection test paper, and the kit including the test strips, the preparation method of the test strips make
The application of the method and the test strips or kit that are detected with the test strips or kit.
Background technique
Rheumatoid factor (RF) is a kind of autoantibody with denatured immunoglobulin G (IgG) for target antigen, and no kind is special
It is anisotropic.Rheumatoid arthritis (RA) patient and Yue 50% healthy human body in have generate RF B cell clone, in certain pathology
Factor such as denaturation IgG or under Epstein-Barr virus directly acts on, can largely be synthetically produced RF.RF and natural IgG binding ability are poor, but easily
With in immune complex IgG or Poly IgG react.RF has 5 seed type of IgG, IgA, IgM, IgD, IgE, the method for detecting RF
Mainly measure IgM class RF.It is significant to the diagnosis of rheumatoid arthritis (RA), parting and observation of curative effect to detect RF.
Immunoglobulin M (IgM) is very important a kind of antibody in the mankind or animal body.When organism infection pathogen
Or after other antigens, in vivo, pathogen or the IgM antibody of other antigentic specificities are occurred that earliest, occurs disease again later
The IgG antibody of substance or other antigentic specificities.It therefore, can be with by the presence or absence of antigen-specific IgM in detection machine body
Diagnose the immune response state of people or animal recent infection pathogen or other antigens.If it is detected that the disease of specificity in serum
Specific IgM caused by substance IgM or other types antigen, can prove that the body has recent pathogenic infection or other antigens
The generation of caused such as autoimmune disease, can be used as the diagnosis and medication foundation of pathogenic infection and immunological diseases etc.,
Patient can be given as soon as possible preferably to treat.Accurately and rapidly diagnostic method can be to avoid antibiotic or other drugs not
Necessity uses, and can provide accurately therapeutic scheme in time, therefore pathogen or other antigen-specific IgM detection methods
Foundation has very important significance.
From the angle analysis of detection immune response principle, antigen-specific IgM is detected usually using prize law and indirect method.
Prize law can generally improve the specificity of detection, but since anti-IgM does not identify specificity and non-specificity IgM, often
It often will cause detection leakage phenomenon, cause detection sensitivity relatively low；And if detected using indirect method principle, it is considered that detection is special
Property IgM antibody before, if not removing the IgG class antibody in clinical samples, it is easy to cause rheumatoid factor (IgM type) and specificity
IgG and the reaction of specific antigen compound lead to IgM false positive, will also result in specific IgG and specific IgM competes antigen knot
Coincidence point and lead to IgM false negative.
The rheumatoid factor or antigen-specific IgM passed through in enzyme linked immunological kit test sample at present has been a kind of
Mature technology.Testing product is divided into three classes substantially:
One kind is traditional micropore panel products, but the time spent by its detection process longer (generally requires several small
When), the reagent and sample size used is also larger, is not able to satisfy a small amount of sample or the clinical demand detected in time, testing principle are
, there is certain deficiency in simple prize law or indirect method.
One kind is fluorescent marker detection slide product, such product reagent dosage is reduced, but detection time is still longer, and
The result of most of such product also needs to observe interpretation by expensive fluorescence microscope, and this intuitive judgment needs technology
The rich experiences of personnel and the time is expended, therefore is not suitable for quickly detection when participating in the cintest.
Another kind of is immunochromatography class product, mostly colloidal gold strip, and than faster and easy, but existing procucts are answered
With there is a certain limitation, such as most products are used only prize law principle and detect IgM antibody, cause omission factor higher, and not
It is able to satisfy multi objective, multisample while detecting.In addition, having less prod using indirect method principle joint-detection, but if the production
Product are not handled and are eliminated to the interference of specific IgG and rheumatoid factor, and false positive and false negative are easy to appear, and are caused
As a result inaccurate.
Immunochromatography technique is a kind of rapid field detection technique, easy to operate, 15 as colloidal gold strip product
It can be visually read in minute as a result, its essential structure is that chromatograph test strip one end on polyvinyl chloride (PVC) backboard is suitable
It is pasted with to secondary mutual overlap joint sample pad, compound pad, nitrocellulose membrane, the other end is pasted with absorption pad.The compound
Pad is the glass fibre element film coated with antibody colloid Au composite (or other particle complex).Have more document reports
IgM antibody is detected using colloidal gold strip, some is handled without the interference to IgG and rheumatoid factor, be easy to cause
As a result inaccurate；Some only simply increases the adsorption line that can adsorb human IgG and rheumatoid factor or adsorption treatment step
Suddenly, the detection line for not increasing rheumatoid factor can not achieve the detection of rheumatoid factor.
But with the growth of the market demand, hospital and patient are badly in need of the product that can combine, quickly detect, this, which also becomes, works as
The trend of modern testing product development.Therefore, interference and principle when being detected based on rheumatoid factor to antigen-specific IgM antibody
Upper analysis, joint, the research and development of quick testing product for carrying out the two are very significant.
Summary of the invention
Therefore, the defect based on above-mentioned prior art, the object of the present invention is to provide a kind of rheumatoid factor (IgM type) and
Other antigen-specific IgM antibody combined detection test papers, and the kit including the test strips, the preparation side of the test strips
Method, the application of the method detected using the test strips or kit and the test strips or kit.
For foregoing invention purpose, the present invention is achieved through the following technical solutions:
The present invention provides a kind of rheumatoid factor (IgM type) and other antigen-specific IgM antibody combined detection test papers,
Wherein, the test strips include the sample pad successively attached on polyvinyl chloride plastic sheet, conjugate pad, nitrocellulose filter and suction
Water cushion, the first detection line (hereinafter may be simply referred to as T1 line) is coated on the nitrocellulose filter, adsorption line (hereinafter can letter
Referred to as A line), one or more detection line (can hereinafter be briefly referred to as T2 line, T3 line ...) and nature controlling line (hereinafter may be used
Referred to as C line)；First detection line is denaturation rabbit/human IgG, and the adsorption line is the antibody of anti-human igg, the nature controlling line
For people IgM, the detection line is respectively selected from one or more of specific antigen: mycoplasma pneumoniae specific antigen, influenza B
The pathogen specifics such as type specific antigen, toxoplasma antigen, autoimmune antigen and anaphylactogen etc..Preferably, the spy
Specific Antigen is to cultivate pathogen after cracking and/or inactivation and pass through the multi-epitope antigen obtained after purification.Wherein, it is related to disease
When mycoplasma antigen is coated with the detection line, the pathogen antigen can be the native purified rear multi-epitope pathogen antigen of extraction,
The multi-epitope pathogen antigen has more highly sensitive than recombinant antigen.
Preferably, according to test strips above-mentioned, wherein the sample pad is glass fibre membrane.And/or the water absorption pad is
Blotting paper.
It is highly preferred that according to test strips above-mentioned, wherein the conjugate pad is to be covered with anti-human IgM antibodies-colloidal gold knot
Close the glass fibre membrane of object.The immunogene of the anti-human IgM antibodies is preferably the Fc5 μ segment of people IgM；And/or it is described anti-human
IgM antibody is preferably the IgG antibody of goat-anti people IgM, 2 segment of F (ab') of the IgG antibody of the more preferably described Yang Yuan.Wherein,
The conjugate pad may be to be covered with other coloured or particle-bound glass fibre membranes of anti-human IgM antibodies-.Wherein, institute
Anti-human IgM antibodies-colloidal gold conjugate is stated with high degree of specificity, the IgG antibody that will not be adsorbed with the adsorption line is handed over
Fork is reacted and is developed the color.The high degree of specificity of the anti-human IgM antibodies-colloidal gold conjugate can also show themselves in that the anti-human IgM
Antibody can remove 2 segment of F (ab') after Fc segment for IgG antibody, which is combining serum sample specificity
After IgM forms compound, rheumatoid factor will not be caused to examine by rheumatoid factor (in conjunction with the Fc section of denaturation IgG) association reaction
The false negative of survey line, to improve the specificity of rheumatoid factor detection line in the present invention.
More preferably, according to test strips above-mentioned, wherein the width of the test strips is convenient width.Preferably 3~
5mm.More preferably 4mm.
It is further preferred that according to test strips above-mentioned, wherein first detection line, adsorption line, detection line and nature controlling line
Convenient width is divided between each adjacent lines.Preferably 3~8mm.More preferably 3mm.Weight between the sample pad and conjugate pad
Folded 1~2mm, preferably 1mm.Chong Die 1~2mm, preferably 1mm between the conjugate pad and nitrocellulose filter.And/or institute
State Chong Die 1~2mm, preferably 1mm between nitrocellulose filter and water absorption pad.
The present invention also provides the preparation methods of above-mentioned test strips, wherein the preparation method includes:
(1) colloidal gold solution is prepared using reduction of sodium citrate gold chloride method: 1000mL tri-distilled water is added into beaker, so
The sodium citrate solution of the 1 mass % of 10mL is added afterwards, after being heated to boiling, the gold chloride for adding the 1 mass % of 10mL is molten
Liquid boils extremely cooling after 15min, obtains the colloidal gold solution；
(2) prepare bonding pad: the colloidal gold solution for taking 20mL step (1) to prepare is placed in a beaker, and 150 μ L 0.1M are added
K2CO3Solution adjusts the pH to 7.6 of solution, stirs evenly, and the anti-human IgM antibodies being centrifuged then are added, and stirs 20min, so
The bovine serum albumen solution of 10 mass % of 2mL is added afterwards, is centrifuged, centrifugation time 40Min, revolving speed 10000rpm, so
After discard supernatant liquid, the bovine serum albumen solution for adding 1 mass % continues to be centrifuged, and liquid is discarded supernatant, using restoring liquid to heavy
Starch is restored, and is coated above the glass fibre membrane of 200 square centimeters of sizes, after -45 DEG C of freezings, as conjugate
Pad, wherein the NaCl that the ingredient for restoring liquid is 150mM, the bovine serum albumen solution of 1 mass %, the sucrose of 0.5 mass %
With the casein-sodium of 0.5 mass %, can be kept in dark place after prepared conjugate pad freeze-drying with room temperature spare；
(3) assemble test strips: conjugate pad, nitrocellulose filter and water absorption pad prepared by sample pad, step (2) is sequentially
It is mutually attached on polyvinyl chloride bottom plate to overlap joint, obtains assembling test strips；
(4) it is coated with nitrocellulose filter: rabbit/human IgG, the antibody of anti-human igg, antigentic specificity will be denaturalized by drawing film instrument
The antigen and people IgM of IgM antibody are successively coated on respectively on the nitrocellulose filter of the assembling test strips of step (3), as institute
State test strips.The test strips can also further progress the operation such as shear, pack, being sealed to prepare corresponding product, example on demand
The PVC offset plate and patch material of test strips are cut into the test strips that width is 4mm as will be described；
Preferably, anti-human IgM antibodies described in step (2) are handled by the following method in advance: using pepsin digestion
The IgG antibody of anti-human IgM, affinity purification processing obtain 2 segment of anti-human IgM antibodies F (ab').
Wherein, glass apparatus used can be impregnated with washing lotion overnight in advance, then be rinsed well.
Preferably, according to the preparation method of test strips above-mentioned, wherein denaturation rabbit/human IgG, anti-human described in step (4)
The concentration of the antibody of IgG, the antigen of antigen-specific IgM antibody and people IgM is each independently 0.3mg/mL.
The present invention also provides a kind of rheumatoid factor (IgM type) and other antigen-specific IgM antibody combined detection reagents
Box, wherein the kit includes above-mentioned test strips or adopts the test strips prepared with the aforedescribed process.Preferably, the examination
Agent box further comprises aluminium foil bag and is sealed in desiccant in the aluminium foil bag.And/or dropper.
It is described the present invention also provides a kind of detection method of rheumatoid factor (IgM type) and other antigen-specific IgMs
Detection method based on above-mentioned test strips, adopt the test strips prepared with the aforedescribed process or above-mentioned kit, wherein the inspection
Survey method includes:
(1) test strips are lain on detection platform, 100 μ l samples is added dropwise to sample pad, the sample is whole blood
Or serum, the colour developing of the first detection line, detection line and nature controlling line is observed after 10~15 minutes as a result, colour developing result after 30 minutes
It is invalid to be considered as；
(2) feelings of (the IgM type) and other antigen-specific IgM antibodies of rheumatoid factor in the sample are judged according to standard
Condition:
When the first detection line, detection line and nature controlling line are displayed in red band, result judgement is；Rheumatoid factor (IgM
Type) positive and other antigen-specific IgM antibodies positive,
When the first detection line is not displayed in red band, detection line and nature controlling line are displayed in red band, and result judgement is；Class
The rheumatism factor (IgM type) is negative and other antigen-specific IgM antibodies are positive,
When the first detection line and nature controlling line are displayed in red band, detection line is not displayed in red band, and result judgement is；Class
The rheumatism factor (IgM type) is positive and other antigen-specific IgM antibodies are negative,
When the first detection line and detection line are not displayed in red band, nature controlling line is displayed in red band, and result judgement is；Class wind
The wet factor (IgM type) is negative and other antigen-specific IgM antibodies are negative,
When nature controlling line is not displayed in red band, result judgement is；Detection is invalid.
Specifically, it is also possible to which following table judges that rheumatoid factor in the sample (IgM type) and other antigen-specific IgMs are anti-
The case where body:
Wherein "+" expression is displayed in red band, and "-" expression is not displayed in red band, and "/" indicates display or is not displayed in red
Band:
The present invention also provides above-mentioned test strips, adopt the test strips prepared with the aforedescribed process or above-mentioned kit exists
Prepare the application in the product for detecting and/or diagnosing one or more of: rheumatoid immunological diseases, it is preferable that described
Rheumatoid immunological diseases are rheumatoid arthritis；Pathogen recent infection, it is preferable that the pathogen is mycoplasma pneumoniae
And/or the multiple pathogens such as Type B influenza virus；Prenatal and postnatal care, it is preferable that the prenatal and postnatal care is detection toxoplasma；Itself exempts from
Epidemic disease systemic disease, it is preferable that the autoimmune pathologies are systemic loupus erythematosus (SLE) and/or xerosis (SS) etc..
In particular it relates to one kind can joint-detection rheumatoid factor (IgM) and other antigen-specific IgMs it is anti-
The method of body.In the method, sample passes sequentially through rheumatoid factor (IgM) detection zone (i.e. T1 line), IgG adsorption zone (i.e. A
Line), other antigen-specific IgM detection zones (i.e. T2 line, T3 line ...).And it provides the method and applies based on immune
Chromatography quickly detects the preparation method in colloidal gold strip.
Particular content is as follows:
Sample pad, conjugate pad, nitrocellulose are sequentially mutually sticked to overlap joint in one end on polyvinyl chloride (PVC) offset plate
Then the material of PVC offset plate and attaching is cut into the test strips of suitable (4mm) width by film, blotting paper.
Wherein, the sample pad is glass fibre membrane, and the conjugate pad is to be covered with anti-human IgM antibodies-colloidal gold to combine
Object or other coloured or particle-bound glass fibre membranes, preferably colloidal gold conjugate；It is coated on the nitrocellulose filter
At least three lines are respectively as follows: T1 line and are denaturalized rabbit/human IgG, for detecting the human rheumatoid factor (IgM)；A line, that is, anti-human igg
Antibody, for adsorbing the IgG antibody in sample；T2 (T3, T4 ...) line, that is, different pathogens or other classes cause antibody response
Specific antigen；C line, that is, people IgM (as shown in Figure 1).
Wherein, the T1 line, that is, human rheumatoid factor (IgM) detection line, T2 (T3, T4 ...) line are the Different Kinds of Pathogens spy optimized
The purified natural multi-epitope specific antigen that Specific Antigen or other class diseases can cause human body IgM antibody to be reacted such as is felt
Infectious pathogen causes the dsDNA of the mycoplasma pneumoniae antigen of pneumonia, autoimmune disease such as systemic loupus erythematosus detection anti-
Former grade, prenatal and postnatal care such as toxoplasma antigen etc. or other any one specific or a variety of spies for IgM antibody detection applications
Specific Antigen.According to detection project demand, it can be achieved that the detection of the specific IgM antibodies of a variety of antigens, and improve sensitivity.
The invention further relates to the analyses that one kind is capable of joint-detection rheumatoid factor (IgM) and other specific IgM antibodies
Method, including colloidal gold strip technology in rheumatoid immunological diseases, pathogen recent infection and autoimmune pathologies or
Application in the specific IgM antibodies detection of other detection applications, particular content are as follows:
After tested sample is added in sample application zone (i.e. sample pad), by chromatography/capillarity, IgM class antibody in sample
It is mobile to the direction of blotting paper one end with the conjugate of anti-human IgM- colloidal gold.If containing rheumatoid factor in tested sample
(IgM type), when sample is moved to the coating line for the IgG that T1 line is denaturalized, rheumatoid factor (IgM type) and anti-human IgM- colloid
The conjugate of gold is captured, shows an aubergine band in T1 line, and rheumatoid factor (IgM type) is no longer moved backward and made
The interference of pairs of specific IgM detection zone.Sample continues to flow, the IgG antibody in sample then with A line, that is, anti-human igg antibody
In conjunction with, for adsorbing the IgG antibody in sample, by specific IgG antibodies capture on A line, no longer with the same antigen that is directed to
The specific IgM competitive reaction site of reaction, the sensitivity and specificity of enhancing specific IgM detection.Sample continues to flow, and contains
Have for the IgM antibody of certain or a variety of primary coated specific antigens, when sample is difference by T2 (T3, T4 ...) line
Specific IgM antibodies and anti-human IgM when pathogen or other classes cause the specific antigen of antibody response to be coated with line, in sample
The conjugate of antibody-colloidal gold is captured, shows that one or more is purplish red in T2 (T3, T4 ...) line by the principle of indirect method
Vitta band, the detection of this detection zone specific IgM no longer sun of vacation caused by by rheumatoid factor that may be present in sample and IgG
The interference of property or false negative.When sample continuously moves to C line, sample carry anti-human IgM antibodies-colloidal gold conjugate then with C
Human IgM antibody at line combines, and shows an aubergine band in C line.
Preferably, being detected sample is whole blood or serum.
If T1, T2 (T3, T4 ...) line is all displayed in red band, illustrate to contain rheumatoid factor (IgM) simultaneously in sample
With certain corresponding species specific IgM；If T1 line is not displayed in red band, and T2 (T3, T4 ...) line is displayed in red band, then
Illustrate containing certain corresponding species specific IgM；If T1 line is displayed in red band, and T2 (T3, T4 ...) line is not displayed in red item
Band then contains rheumatoid factor (IgM)；If T1 and T2 (T3, T4 ...) line do not develop the color, then it represents that rheumatoid factor (IgM) and
Certain specific IgM is feminine gender；No matter T1, T2 (T3, T4 ...) line what state, as long as not being displayed in red band at C line,
Then the test strips are determined as in vain.
If specific IgM is the positive, show that the body has certain pathogen of recent infection, or have autoimmune antibody different
Paradoxical reaction；If RF (IgM type) is the positive, shows that the body there may be the abnormal factors for causing rheumatoid class to generate, prompt
There are the risks of rheumatoid immunity disease；If RF (IgM type) and specific IgM are all the positives, show that the body is felt in the recent period
Certain pathogen is contaminated, and presence there may be the abnormal factors for causing wet class of class point to generate；If RF (IgM type) and specificity
IgM is feminine gender, shows not infect certain pathogen, and the abnormal factors risk that wet class of class point generates is lower；No matter T1, and
What state at T2 (T3, T4 ...), if C line does not develop the color, which is considered as invalid.
Test strips provided by the invention are based on this sample and pass sequentially through rheumatoid factor (IgM) detection zone, IgG absorption
Area, other antigen-specific IgM detection zones analysis method, change traditional independent detection specific IgM antibodies or rheumatoid
The method of the factor (IgM type) overcomes in conventional detection rheumatoid factor to be easy to detect specific IgM antibodies and interfere
Deficiency, and joint-detection excludes to be by whether there is the testing result of rheumatoid factor (IgM type) in the same sample
It is no to have rheumatoid factor that detect the influence for causing false positive to interfere to subsequent specific IgM antibodies, if a such as sample
The testing result for detecting rheumatoid factor (IgM type) is feminine gender, then can excluding the sample first, there is no rheumatoid factor (IgM
Type) to subsequent specific IgM antibodies detection interference；If the testing result of a pattern detection rheumatoid factor (IgM type)
For the positive, then show that our detection method first specifies that this is that there are rheumatoid factor (IgM type) to specific IgM for a regular meeting
The sample that antibody test interferes, but the rheumatoid captured in sample is detected since our design increases this
The factor (IgM type), therefore avoid to the interference test strips of subsequent specific IgM antibodies detection in specific antigen IgM
Before antibody detection line, increase rheumatoid factor
(IgM) detection line and absorption IgG line, can not only detect a variety of specific IgM antibodies simultaneously in one test
And the interference of IgG is removed, it is detected while also achieving rheumatoid factor (IgM type), and subtracted rheumatoid factor in sample
It interferes with IgG antibody using false positive caused by indirect method detection specific IgM antibodies, has fully demonstrated in this detection scheme
Indirect method principle detects the advantage of the sensitivity and specificity of specific IgM.
Based on immunochromatography technique, the colloidal gold immunochromatographimethod technology which more highlights multi objective is quickly examined
It surveys, is cheap, reading the features such as result is quick and intuitive, not needing special instruments and equipment, also do not need the behaviour of professional
Make, the overall coincidence rate of testing result is higher, can greatly shorten detection time and reduce testing cost, reach on-site test
Purpose.
In addition to this, which, which improves, commonly uses prize law principle detection specific IgM in previous immunochromatography technique
The deficiency of antibody because cannot identify specific IgM and non-specificity IgM using the detection zone of prize law principle, and may be made
At only capture non-specificity IgM, and the case where specific IgM missing inspection, so that sensitivity decrease；Next improves immune layer in the past
Analysis technology is coated with recombinant antigen using indirect method principle more, and the case where lead to that epitope is single and make sensitivity decrease, this
The multi-epitope antigen of the pathogen of method coating optimization naturally cultivated through subsequent processing and after purification, improves sensitivity；?
Previous immunochromatography technique is improved using indirect method principle, but the detection method without removing IgG and rheumatoid factor influence,
And detection specific IgM antibodies is caused the case where false positive and false negative occur, though or have at IgG absorption in the indirect method that has
Reason, but it is not carried out rheumatoid factor
(IgM) it is detected while, cannot define and excludes certain sample and whether really there is rheumatoid factor IgM is caused to detect
The interpretation of interference.In other words, test strips provided by the invention can either avoid IgG antibody and class wind in sample in one test
The interference of wet factor pair detection, and indirect method can be utilized to improve sensitivity, and realization while detection and the clear (area) dtex opposite sex
The feminine gender and positive findings of IgM and rheumatoid factor (IgM), so that more relevant informations are obtained, with important clinical meaning
Justice.
Detailed description of the invention
Hereinafter, carrying out the embodiment that the present invention will be described in detail in conjunction with attached drawing, in which:
Fig. 1 shows the signal of a kind of rheumatoid factor (IgM type) and the detection method of other antigen-specific IgM antibodies
Figure；
Fig. 2 shows rheumatoid factor (IgM type) and other antigen-specific IgM antibody combined detection test papers to illustrate
Figure；
Fig. 3 shows Fig. 3~5 label used and the test strips structure；
Fig. 4 shows the effective testing result of the test strips；
Fig. 5 shows the invalid testing result of the test strips；
Description of symbols:
1, sample application zone；2, anti-human IgM antibodies-particle-bound area；3, nitrocellulose filter；4, RF (IgM type) detection zone；
5, IgG antibody area is adsorbed；6, specific IgM antibodies detection zone；7, quality inspection area；8, absorption pad；
9, sample pad；10, conjugate pad (can be and be covered with anti-human IgM antibodies-colloidal gold conjugate conjugate pad)；
11, T1 line (i.e. detection rheumatoid factor (IgM type) area)；12, A line (i.e. absorption IgG antibody area)；13, T2 line is (i.e. specific
IgM antibody detection zone)；14, C line (i.e. quality control region)；15, water absorption pad (can be blotting paper)；
16, it is denaturalized IgG (can be denaturation rabbit/human IgG)；17, the antibody of anti-human igg；18, rheumatoid factor (can be
Rheumatoid factor (IgM type))；19, specific IgM；20, people IgM；21, the antigen of specific IgM；22, gold labeling antibody conjugate
(can be anti-human IgM antibodies-colloidal gold conjugate)；23, nano particle (can be colloidal gold)；24, anti-human IgM antibodies；25,
Human IgG；
26, testing result is that RF (IgM) is positive and specific antibody IgM is positive；27, testing result is RF (IgM) negative
With the specific antibody IgM positive；28, testing result is that RF (IgM) is positive and specific antibody IgM is negative；29, testing result is
RF (IgM) is negative and specific antibody IgM is negative；
It 30~33, is respectively four kinds of invalid situations of testing result.
Specific embodiment
Present invention will be further explained by specific examples below, it should be understood, however, that, these embodiments are only
It is used, is but should not be understood as present invention is limited in any form for specifically describing in more detail.
In following embodiment, in addition to the test material, condition and the operating method that particularly point out, used in embodiment
Many materials and operating method are well known in the art.Therefore, it will be apparent to those skilled in the art that within a context, if not special
Do not mentionlet alone bright, material therefor of the present invention and operating method are well known in the art.
Reagent and instrument used in the following embodiment are as follows:
Reagent:
It is denaturalized rabbit/human IgG: Fei Peng Biological Co., Ltd.；
The antibody of the antibody of anti-human IgM, anti-human igg: Beijing Bo Aosen Bioisystech Co., Ltd；
People IgM:sigma；
Nitrocellulose filter: Merck Mi Libo；
PVC offset plate, blotting paper, glass fibre membrane: Shanghai Jinbiao Bio-Tech Co., Ltd.；
Mycoplasma pneumoniae IgM antibody detection kit (enzyme-linked immunization): Anqun Bioengineering Co., Ltd., Shenzhen；
Rheumatoid factor IgM antibody detection kit (colloidal gold method): Weifang Kanghua Biotech. Co., Ltd..
Instrument:
Three-dimensional planar draws film instrument: Shanghai Jinbiao Bio-Tech Co., Ltd.；
Freeze drier: Beijing Bo Yikang laboratory apparatus Co., Ltd；
Cutting machine: Shanghai Jinbiao Bio-Tech Co., Ltd..
Embodiment 1
The verification method and result for the high degree of specificity anti-human IgM antibodies that the present embodiment is used to illustrate that the present invention selects.
Method 1 includes: the human IgG of purifying, and night packet is spent in polystyrene micropore plate 4 in the concentration hole 1ug/ml, 100ul/
Quilt；Using 3% bovine serum albumin(BSA) (BSA), the hole 200ul/, 37 degree are closed for 2 hours；Then anti-human IgM-HRP is added,
The hole 100ul/, 37 degree 30 minutes, after washing, be added TMB developing solution, be protected from light after ten minutes carry out microplate reader detection OD450nm-630nm；
And do the Positive control wells and blank control wells of coating people IgM；It is porous-parallel, averages.Testing result such as table 1.
Method 2 includes: that test strips include the sample pad successively attached on polyvinyl chloride plastic sheet, conjugate pad, nitric acid fibre
Tie up plain film and water absorption pad；The sample pad is glass fibre membrane；The conjugate pad is with anti-human IgM antibodies-colloidal gold knot
Close the glass fibre membrane of object；It is successively coated with detection line and nature controlling line respectively on the nitrocellulose filter；The detection line is
Human IgG, the nature controlling line behaviour IgM；The water absorption pad is blotting paper.To sample pad be added dropwise 100 μ l pbs buffers, 10~15
The colour developing of detection line and nature controlling line is observed after minute as a result, the colour developing result after 30 minutes is considered as in vain.Testing result such as table 2.
The testing result of method 1 and method 2 from the present embodiment is it is found that the anti-human IgM antibodies that the present invention uses have height
Degree specificity, not with human IgG cross reaction.
Table 1
Table 2
Embodiment 2
The present embodiment is for illustrating rheumatoid factor of the invention (IgM type) and other antigen-specific IgM antibodies joint
Test strip and kit and preparation method thereof.
As shown in Fig. 2, the test strips of the present embodiment include the sample pad successively attached on polyvinyl chloride plastic sheet, conjugate
Pad, nitrocellulose filter and water absorption pad；The sample pad is glass fibre membrane；The conjugate pad is with anti-human IgM antibodies-
The glass fibre membrane of colloidal gold conjugate；The first detection line, adsorption line, one are successively coated on the nitrocellulose filter respectively
Detection line and nature controlling line；First detection line is denaturation rabbit/human IgG, and the adsorption line is the antibody of anti-human igg, described
Nature controlling line behaviour IgM, the detection line are by the natural multi-epitope antigen of mycoplasma pneumoniae specificity after purification；The water suction
Pad is blotting paper.
Wherein, 3~8mm is divided between each adjacent lines of the first detection line, adsorption line, detection line and nature controlling line.The sample
Chong Die 1~2mm between product pad and conjugate pad, Chong Die 1~2mm between the conjugate pad and nitrocellulose filter, the nitre
Chong Die 1~2mm between acid cellulose film and water absorption pad.The width of the test strips is convenient width.In the present embodiment, first
3mm is divided between detection line, adsorption line, detection line and each adjacent lines of nature controlling line.Weight between the sample pad and conjugate pad
Folded 1mm, be overlapped 1mm between the conjugate pad and nitrocellulose filter are be overlapped between the nitrocellulose filter and water absorption pad
1mm, the width of the test strips are 4mm.
Above-mentioned test strips the preparation method is as follows:
(1) colloidal gold solution is prepared using reduction of sodium citrate gold chloride method: 1000mL tri-distilled water is added into beaker, so
The sodium citrate solution of the 1 mass % of 10mL is added afterwards, after being heated to boiling, the gold chloride for adding the 1 mass % of 10mL is molten
Liquid boils extremely cooling after 15min, obtains the colloidal gold solution；
(2) prepare bonding pad: the colloidal gold solution for taking 20mL step (1) to prepare is placed in a beaker, and 150 μ L 0.1M are added
K2CO3Solution adjusts the pH to 7.6 of solution, stirs evenly, and the anti-human IgM antibodies being centrifuged then are added, and stirs 20min, so
The bovine serum albumen solution of 10 mass % of 2mL is added afterwards, is centrifuged, centrifugation time 40Min, revolving speed 10000rpm, abandons
Supernatant is removed, the bovine serum albumen solution for adding 1 mass % continues to be centrifuged, and liquid is discarded supernatant, using recovery liquid to sediment
Restored, is coated above the glass fibre membrane of 200 square centimeters of sizes, after -45 DEG C of freezings, as conjugate pad,
The NaCl spare, the ingredient for restoring liquid is 150mM, the cow's serum egg of 1 mass % is kept in dark place at room temperature after freeze drying
White solution, the casein-sodium of the sucrose of 0.5 mass % and 0.5 mass %；
(3) assemble test strips: conjugate pad, nitrocellulose filter and water absorption pad prepared by sample pad, step (2) is sequentially
It is mutually attached on polyvinyl chloride bottom plate to overlap joint, adjacent part is overlapped 1mm, obtains assembling test strips；
(4) be coated with nitrocellulose filter: rabbit/human IgG, the antibody of anti-human igg, lung will be denaturalized by drawing film instrument using three-dimensional planar
On the nitrocellulose filter for the assembling test strips that scorching mycoplasma specific antigen and people IgM are successively coated on step (3) respectively,
In draw film instrument setup parameter are as follows: scribing line interval be 3mm, scribing line concentration be 0.3mg/mL；
(5) by step (4), treated that assembling test paper is cut into the test strips of width 4mm.The present embodiment can also be further
There is provided kit comprising the test strips of aluminium foil bag and the desiccant being sealed in the aluminium foil bag, dropper and above-mentioned preparation.
The kit the preparation method comprises the following steps: by the test strips of above-mentioned preparation in the aluminium foil bag, be reloaded into a desiccant
And dropper, it is sealed and saves at room temperature.
Embodiment 3
The test strips or kit that the present embodiment is used to illustrate to prepare using embodiment 2 are former in RF (IgM type) and pneumonia branch
Application in body pattern detection.
Steps are as follows for the application method of the test strips:
(1) blood serum sample is collected；
(2) test strips are lain on detection platform, to sample pad be added dropwise 100 μ l steps (1) blood serum sample, 10~15
The colour developing of the first detection line, detection line and nature controlling line is observed after minute as a result, the colour developing result after 30 minutes is considered as in vain；
(3) result judges: determining that "-" expression is not displayed in red as a result, wherein "+" expression is displayed in red band according to table 3
Band, "/" indicate display or are not displayed in red band:
Table 3
The test paper slip prepared using embodiment 2 detects the clinical positive serum made a definite diagnosis and negative serum,
It the results are shown in Table 4.
4 embodiment of table, 2 testing result
Embodiment 4
The present embodiment is for illustrating rheumatoid factor of the invention (IgM type) and other antigen-specific IgM antibodies joint
Test strip and kit and preparation method thereof.
As shown in Fig. 2, the test strips of the present embodiment include including the sample pad successively attached on polyvinyl chloride plastic sheet, knot
Close object pad, nitrocellulose filter and water absorption pad；The sample pad is glass fibre membrane；The conjugate pad is with anti-human IgM
The glass fibre membrane of antibody-colloidal gold conjugate；The first detection line, absorption are successively coated on the nitrocellulose filter respectively
Line, two detection lines and nature controlling line；First detection line is denaturation rabbit/human IgG, and the adsorption line is the antibody of anti-human igg,
The nature controlling line behaviour IgM, the detection line are respectively by the natural multi-epitope antigen of mycoplasma pneumoniae specificity after purification
With the natural multi-epitope antigen of influenza Type B specificity of process after purification；The water absorption pad is blotting paper.
Wherein, 3mm is divided between each adjacent lines of the first detection line, adsorption line, detection line and nature controlling line.The sample pad
The be overlapped 1mm between conjugate pad, be overlapped 1mm between the conjugate pad and nitrocellulose filter, the nitrocellulose filter
The be overlapped 1mm between water absorption pad.The width of the test strips is 4mm.
Above-mentioned test strips the preparation method is as follows:
(1) colloidal gold solution is prepared using reduction of sodium citrate gold chloride method: 1000mL tri-distilled water is added into beaker, so
The sodium citrate solution of the 1 mass % of 10mL is added afterwards, after being heated to boiling, the gold chloride for adding the 1 mass % of 10mL is molten
Liquid boils extremely cooling after 15min, obtains the colloidal gold solution；
(2) prepare bonding pad: the colloidal gold solution for taking 20mL step (1) to prepare is placed in a beaker, and 150 μ L 0.1M are added
K2CO3Solution adjusts the pH to 7.6 of solution, stirs evenly, and the anti-human IgM antibodies being centrifuged then are added, and stirs 20min, so
The bovine serum albumen solution of 10 mass % of 2mL is added afterwards, is centrifuged, centrifugation time 40Min, revolving speed 10000rpm, abandons
Supernatant is removed, the bovine serum albumen solution for adding 1 mass % continues to be centrifuged, and liquid is discarded supernatant, using recovery liquid to sediment
Restored, is coated above the glass fibre membrane of 200 square centimeters of sizes, after -45 DEG C of freezings, as conjugate pad,
The NaCl spare, the ingredient for restoring liquid is 150mM, the cow's serum egg of 1 mass % is kept in dark place at room temperature after freeze drying
White solution, the casein-sodium of the sucrose of 0.5 mass % and 0.5 mass %；
(3) assemble test strips: conjugate pad, nitrocellulose filter and water absorption pad prepared by sample pad, step (2) is sequentially
It is mutually attached on polyvinyl chloride bottom plate to overlap joint, adjacent part is overlapped 1mm, obtains assembling test strips；
(4) be coated with nitrocellulose filter: rabbit/human IgG, the antibody of anti-human igg, lung will be denaturalized by drawing film instrument using three-dimensional planar
Scorching mycoplasma specific antigen, influenza Type B specific antigen and people IgM are successively coated on the assembling test strips of step (3) respectively
Nitrocellulose filter on, wherein drawing the setup parameter of film instrument are as follows: scribing line interval be 3mm, scribing line concentration be 0.3mg/mL；
(5) by step (4), treated that assembling test paper is cut into the test strips of width 4mm.The present embodiment can also be further
There is provided kit comprising the test strips of aluminium foil bag and the desiccant being sealed in the aluminium foil bag, dropper and above-mentioned preparation.
The kit the preparation method comprises the following steps: by the test strips of above-mentioned preparation in the aluminium foil bag, be reloaded into a desiccant
And dropper, it is sealed and saves at room temperature.
Embodiment 5
The test strips or kit that the present embodiment is used to illustrate to prepare using embodiment 4 are former in RF (IgM type) and pneumonia branch
Application in body, influenza Type B pattern detection.
Steps are as follows for the application method of the test strips:
(1) blood serum sample is collected；
(2) test strips are lain on detection platform, to sample pad be added dropwise 100 μ l steps (1) blood serum sample, 10~15
The colour developing of the first detection line, detection line and nature controlling line is observed after minute as a result, the colour developing result after 30 minutes is considered as in vain；
(3) result judges: determining result according to table 3.
The test paper slip prepared using embodiment 4 detects the clinical positive serum made a definite diagnosis and negative serum,
Wherein, RF (IgM type) and mycoplasma pneumoniae IgM antibody positive sample are identical with case study on implementation 2, and it is special to newly increase influenza Type B
Property IgM positive sample 100 and the common negative sample of three index IgM 150 are detected.
It the results are shown in Table 5.
5 embodiment of table, 5 testing result
Test example 1
This test example is used to illustrate the detection comparative test of test strips and commercially available testing product prepared by embodiment 2.
Given the test agent be 100 clinics made a definite diagnosis rheumatoid factor (IgM type) positive sample, 100 clinics
The mycoplasma pneumoniae positive sample made a definite diagnosis and 150 total negative samples.
Test strips prepared by embodiment 2 are detected using the method for embodiment 3.
Mycoplasma pneumoniae IgM antibody detection kit (enzyme-linked immunization) and rheumatoid factor IgM antibody detection kit
(colloidal gold method) is detected using application method in its specification.
Testing result is shown in Table 6.From testing result it is found that with mycoplasma pneumoniae IgM antibody detection kit (enzyme linked immunological
Method) it compares, test strips prepared by embodiment 2 have higher coincidence rate to given the test agent；And it is examined with rheumatoid factor IgM antibody
Test agent box (colloidal gold method) is compared, and test strips prepared by embodiment 2 are stable to the testing result of rheumatoid factor and more quasi-
Really.
Table 6
Although here, having carried out a degree of description to the present invention, it will be apparent that, do not depart from spirit of the invention and
Under conditions of range, one of ordinary skill in the art can carry out the appropriate variation of each condition.It is understood that the present invention is unlimited
Summarize in the embodiment and specific example, right are attributed to the scope of the claims, and including each factor etc.
With replacement.
Claims (23)
1. a kind of IgM type rheumatoid factor and other antigen-specific IgM antibody combined detection test papers, which is characterized in that institute
Stating test strips includes the sample pad successively attached on polyvinyl chloride plastic sheet, conjugate pad, nitrocellulose filter and water absorption pad, institute
It states and is successively coated with the first detection line, adsorption line, one or more detection line and nature controlling line on nitrocellulose filter respectively；It is described
First detection line is to be denaturalized rabbit and/or transsexual person IgG, antibody of the adsorption line for anti-human igg, the nature controlling line behaviour IgM,
The detection line is respectively selected from one or more of specific antigen: mycoplasma pneumoniae specific antigen, influenza Type B specificity
Antigen, toxoplasmosis pathogen-specific antigen, autoimmune antigen and anaphylactogen；The conjugate pad is to be covered with anti-human IgM
The glass fibre membrane of antibody-colloidal gold conjugate, the Fc5 μ segment of the immunogene behaviour IgM of the anti-human IgM antibodies；It is described anti-
Human IgM antibody is the IgG antibody of goat-anti people IgM.
2. test strips according to claim 1, which is characterized in that the specific antigen is culture pathogen through cracking
And/or after inactivation and pass through the multi-epitope antigen obtained after purification.
3. test strips according to claim 1, which is characterized in that the sample pad is glass fibre membrane；And/or the suction
Water cushion is blotting paper.
4. test strips according to claim 1, which is characterized in that the IgG antibody of the goat-anti people IgM is the IgG of Yang Yuan
2 segment of F (ab') of antibody, and the width of the test strips is 3~5mm.
5. test strips according to claim 1-3, which is characterized in that the width of the test strips is 3~5mm.
6. test strips according to claim 5, which is characterized in that the width of the test strips is 4mm.
7. test strips according to claim 1-4, which is characterized in that
Convenient width is divided between first detection line, adsorption line, detection line and each adjacent lines of nature controlling line；
Chong Die 1~2mm between the sample pad and conjugate pad；
Chong Die 1~2mm between the conjugate pad and nitrocellulose filter；And/or
Chong Die 1~2mm between the nitrocellulose filter and water absorption pad.
8. test strips according to claim 7, which is characterized in that first detection line, adsorption line, detection line and Quality Control
3~8mm is divided between each adjacent lines of line.
9. test strips according to claim 8, which is characterized in that first detection line, adsorption line, detection line and Quality Control
3mm is divided between each adjacent lines of line.
10. test strips according to claim 7, which is characterized in that be overlapped 1mm between the sample pad and conjugate pad.
11. test strips according to claim 7, which is characterized in that weight between the conjugate pad and nitrocellulose filter
Folded 1mm.
12. test strips according to claim 7, which is characterized in that be overlapped between the nitrocellulose filter and water absorption pad
1mm。
13. the preparation method of test strips described in any one of claims 1 to 12, which is characterized in that the preparation method packet
It includes:
(1) colloidal gold solution is prepared using reduction of sodium citrate gold chloride method: 1000mL tri-distilled water being added into beaker, then plus
Enter the sodium citrate solution that the mass percent concentration of 10mL is 1%, after being heated to boiling, adds the mass percent of 10mL
The chlorauric acid solution that concentration is 1% boils extremely cooling after 15min, obtains the colloidal gold solution；
(2) prepare conjugate pad: the colloidal gold solution for taking 20mL step (1) to prepare is placed in a beaker, and is added 150 μ L0.1M's
K2CO3Solution adjusts the pH to 7.6 of solution, stirs evenly, and the anti-human IgM antibodies being centrifuged then are added, and stirs 20min, then
The bovine serum albumen solution that 2mL mass percent concentration is 10% is added, is centrifuged, centrifugation time 40min, revolving speed is
10000rpm discards supernatant liquid, adds the bovine serum albumen solution that mass percent concentration is 1% and continues to be centrifuged, discards
Clear liquid restores sediment using liquid is restored, is coated above the glass fibre membrane of 200 square centimeters of sizes, -45
After DEG C freezing, as conjugate pad, described to restore liquid be ingredient is the ox blood that the NaCl of 150mM, mass percent concentration are 1%
The solution for the casein-sodium that the sucrose and mass percent concentration that albumin, mass percent concentration are 0.5% are 0.5%；
(3) assemble test strips: conjugate pad, nitrocellulose filter and water absorption pad prepared by sample pad, step (2) is sequentially mutual
It is attached on polyvinyl chloride plastic sheet to overlap joint, obtains assembling test strips；
(4) be coated with nitrocellulose filter: using drawing, film instrument will be denaturalized rabbit and/or transsexual person IgG, the antibody of anti-human igg, antigen are special
The antigen and people IgM of anisotropic IgM antibody are successively coated on respectively on the nitrocellulose filter of the assembling test strips of step (3).
14. the preparation method of test strips according to claim 13, which is characterized in that anti-human IgM described in step (2) is anti-
Body is handled by the following method in advance: using the IgG antibody of the anti-human IgM of pepsin digestion, affinity purification is handled described in acquisition
Anti-human IgM antibodies.
15. the preparation method of test strips according to claim 13, which is characterized in that described in step (4) denaturation rabbit and/
Or antibody, the antigen of antigen-specific IgM antibody and the concentration of people IgM of transsexual person IgG, anti-human igg are each independently
0.3mg/mL。
16. a kind of IgM type rheumatoid factor and other antigen-specific IgM antibody combined detection kits, which is characterized in that institute
Stating kit includes: any one of test strips or use claim 13 to 15 as described in any one of claims 1 to 12
The test strips of the method preparation.
17. kit according to claim 16, which is characterized in that the kit further comprises:
Aluminium foil bag and the desiccant being sealed in the aluminium foil bag；And/or
Dropper.
18. test strips described in any one of claims 1 to 12, using method described in any one of claim 13 to 15
Kit described in the test strips or claim 16 or 17 of preparation preparation for detecting and/or diagnose selected from following a kind of or
Application in a variety of products:
Pathogen recent infection；
Prenatal and postnatal care；
And autoimmune pathologies.
19. application according to claim 18, which is characterized in that the autoimmune pathologies are that disease is immunized in rheumatoid
Disease.
20. application according to claim 19, which is characterized in that the rheumatoid immunological diseases are rheumatoid joint
It is scorching.
21. application according to claim 18, which is characterized in that the pathogen is mycoplasma pneumoniae and/or Type B influenza
Virus.
22. application according to claim 18, which is characterized in that the prenatal and postnatal care is detection toxoplasma.
23. application according to claim 18, which is characterized in that the autoimmune pathologies are systemic red yabbi
Sore and/or xerosis.
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| CN107300616B (en) * | 2017-06-20 | 2019-02-01 | 广东云天抗体生物科技有限公司 | A kind of freeze-drying formula of liquid and its application |
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| DE19649389A1 (en) * | 1996-11-29 | 1998-06-04 | Boehringer Mannheim Gmbh | Antigen-specific IgM detection |
| ITMI20012160A1 (en) * | 2001-10-18 | 2003-04-18 | Edoardo Marchisio | IMMUNOENZYMATIC SYSTEM FOR THE ANTIGENIC CHARACTERIZATION OF THE ACTIVE INFECTION BY CYTOMEGALOVIRUS AND CMV CONFIRMATION TEST |
| CN101221177B (en) * | 2008-01-25 | 2011-06-08 | 马义才 | Portable fast joint inspection device for foetus teratogenic disease |
| CN101706503A (en) * | 2009-07-08 | 2010-05-12 | 无锡博慧斯生物医药科技有限公司 | Human IgG and rheumatoid factor absorption device and related detection method of IgM antibody in human serum |
| CN102109520A (en) * | 2009-12-29 | 2011-06-29 | 北京库尔科技有限公司 | Herpes simplex virus I and II IgM antibody joint inspection kit and preparation method thereof |
| WO2011125877A1 (en) * | 2010-03-31 | 2011-10-13 | 積水メディカル株式会社 | Measurement method using immunochromatography, test strip for immunochromatography, and measurement reagent kit for immunochromatography |
| CN102507932A (en) * | 2011-12-02 | 2012-06-20 | 无锡博慧斯生物医药科技有限公司 | IgM (immunoglobulin M) antibody detection test strip |
| CN102707050A (en) * | 2012-05-24 | 2012-10-03 | 蓝十字生物药业（北京）有限公司 | Colloidal gold test strip for fast detecting rheumatoid factors |
| CN102749446A (en) * | 2012-07-21 | 2012-10-24 | 北京中检安泰诊断科技有限公司 | Chlamydia pneumoniae IgM (immunoglobulin M) colloidal golden method kit and preparation method thereof |
| CN202994798U (en) * | 2012-10-29 | 2013-06-12 | 上海科新生物技术股份有限公司 | Diagnostic kit for diagnosis of rheumatoid arthritis |
| CN203772871U (en) * | 2014-03-12 | 2014-08-13 | 武汉中博生物股份有限公司 | Colloidal gold immunochromatography detection test paper for porcine circovirus type 2 antibody and porcine reproductive and respiratory syndrome virus IgM (Immunoglobulin M) antibody |
| CN205038218U (en) * | 2015-09-21 | 2016-02-17 | 广州万孚生物技术股份有限公司 | Instant detect reagent card and kit |
| CN206362808U (en) * | 2016-07-20 | 2017-07-28 | 国家纳米科学中心 | IgM types rheumatoid factor and antigen-specific antibodies joint inspection test paper and kit |
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