CN108761067B - Biomarkers for detection of idiopathic inflammatory myopathy and uses thereof - Google Patents
Biomarkers for detection of idiopathic inflammatory myopathy and uses thereof Download PDFInfo
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Abstract
The invention discloses a use of signal peptide recognition particle 19, namely SRP19, or a fragment thereof in the preparation of an agent for diagnosing idiopathic inflammatory myopathy. The invention adopts the high-density protein chip technology to screen the target antigen of the PM/DM related autoantibody, and 91 PM/DM related autoantibodies are determined. Adopting 91 PM/DM related autoantibodies to correspond to a target antigen to construct and prepare a PM/DM related autoantibody target antigen chip, and carrying out clinical amplification sample verification. The results showed that the anti-SRP 19 antibody had 19.00% positive rate in the PM group, which was 2.67% higher than the positive rate in all the control groups (P ═ 3.26 × 10)‑7) The difference in results was significant; the positive rate in DM group is 7.41%, which is higher than that in all control group by 2.67% (P ═ 0.80X 10)‑2) The difference in results was significant; the positive rate in the PM/DM group is 10.33%, which is higher than that in all the control groups by 2.67% (P ═ 0.09X 10)‑3) The difference in results was significant. Indicating that anti-SRP 19 antibodies are useful as diagnostic and detection markers for idiopathic inflammatory myopathy.
Description
Technical Field
The invention belongs to the field of biological detection, and particularly relates to a biomarker for detecting polymyositis and/or dermatomyositis and application thereof.
Background
Idiopathic Inflammatory Myopathies (IIMs) are heterogeneous diseases characterized by proximal symmetric muscle weakness and multiple organ involvement, and mainly include clinical subtypes such as Polymyositis (PM), Dermatomyositis (DM), immune-mediated necrotizing myopathy (IMNM), sporadic inclusion body myositis (sIBM), and Juvenile Idiopathic Myositis (JIM). PM/DM is a striated muscle non-suppurative inflammatory myopathy autoimmune disease. The clinical characteristics are that serum creatinase is increased, electromyogram is changed in myogenesis, and muscle tissues such as limb girdle muscle, cervical muscle, pharyngeal muscle and the like are subjected to inflammation and degeneration change, so that symmetric muscle weakness and certain muscle atrophy are caused, and finally a plurality of systems and organs are involved, so that a patient dies. PM/DM is not uncommon in China, women are more common, and the proportion of men to women is about 1: 2. the disease can occur at any age, is bimodal, has a peak at 5-14 years old and 45-60 years old of adults, is easy to have Interstitial Lung Disease (ILD), and the complication is an important influencing factor influencing the PM/DM prognosis.
Autoantibodies are antibodies directed against tissues, organs, cells and cell components of the body, and are important weapons of the humoral immune system of the body to defend against pathogenic bacteria invasion and maintain the growth balance of the body. Serum autoantibody markers are produced during the pathogenesis of autoimmune diseases and often occur years or even more before the disease has a typical clinical presentation and any laboratory or imaging abnormalities. Evidence suggests that the onset of PM/DM is associated with autoimmunity: (1) histopathological examination showed that the patient's muscle tissue appeared to present T cells, macrophages, dendritic cells, B cells and plasma cells; (2) serum detection revealed the presence of autoantibodies to nuclear cytosolic components in PM/DM patients. Autoantibodies associated with myositis are classified into two categories based on their diagnostic value for disease: myositis-specific antibodies (MSAs) and myositis-associated antibodies (MAAs). MSAs have been reported to have anti-ARS antibodies (anti-Jo-1, PL-7, PL-12, EJ, OJ, KS, YRS, Zo antibodies), anti-Mi-2 antibodies, anti-MDA 5 antibodies, anti-TIF 1 antibodies, anti-NXP 2 antibodies, anti-SAE antibodies, anti-SRP antibodies, anti-HMGCR antibodies, anti-cN 1A antibodies; MAAs include anti-PM-Scl antibody, anti-Ku antibody, anti-SSA antibody, anti-Ro 50 antibody, anti-U1-RNP antibody, anti-SSB antibody, and AECA. MSAs may be associated with specific clinical subtypes, aid in predicting complications, aid in diagnosis, prognosis and prompt for clinical selection of the correct treatment strategy.
anti-ARS antibody positive myositis patients are usually diagnosed with anti-synthetase antibody syndrome (ASS). Such patients have a particular set of symptoms: ILD, raynaud's phenomenon, craftsman's hands, non-erosive arthritis, fever or may be accompanied by rashes. Most common is an anti-Jo-1 antibody, which may be present in 9% -24% of IIMs. The anti-Mi-2 antibody is a DM specific antibody, the target antigen of the antibody is a nucleosome remodeling and deacetylase (NuRD) complex, the antibody has a positive rate of 11-59% in adult DM and 4-10% in Juvenile Dermatomyositis (JDM), and the patient with myositis positive to the anti-Mi-2 antibody has relatively light illness, can have clinical manifestations of arthralgia, arthritis, Raynaud phenomenon, ILD and the like, and has relatively good treatment response and prognosis. The anti-MDA 5 antibody is a DM-specific antibody, the target antigen of which is melanoma differentiation associated protein 5, most commonly found in patients with clinical myopathic dermatomyositis (CADM), and is closely associated with clinical manifestations: the anti-MDA 5 antibody is obviously related to skin phenotypic characteristics such as skin ulceration, touchable palmar pimples and the like, can increase the risks of oral pain/ulcer, hand swelling, arthritis/joint pain and diffuse alopecia, and has higher diagnostic value on Rapid Progressive Interstitial Lung Disease (RPILD) related to myositis. The anti-TIF 1 antibody is a DM specific antibody, and the target antigen is transcription mediator 1 family protein, including three subtypes of anti-TIF 1 alpha, anti-TIF 1 beta and anti-TIF 1 gamma antibodies. anti-TIF 1 antibody was found in both adult and pediatric patients with adult PM/DM frequency of 13-31% and JDM frequency of 22-29%. Since the TIF1 protein plays a key role in tumorigenesis, these antibodies may be generated due to abnormal anti-tumor immunity in vivo, and anti-TIF 1 antibodies are most commonly found in tumor-associated myositis (CAM). The anti-NXP 2 antibody mainly appears in serum of a patient with juvenile myositis, the target antigen of the anti-NXP 2 antibody is nuclear matrix protein 2, the positivity of the antibody is 23% -25%, the positivity of the anti-NXP 2 antibody in a patient with adult myositis is 1% -17%, the JDM patient positive for the anti-NXP 2 antibody is characterized by muscle contracture, atrophy and remarkable damage to muscle function, and the antibody is related to calcareosis. The anti-SAE antibody target antigen is a small ubiquitin-like modifier-1 (SUMO-1) activator heterodimer: SAE1 and SAE2, autoantibodies mainly associated with DM, most positive patients first develop skin lesions and subsequently can develop severe dysphagia. anti-SRP antibodies target antigens for signal peptide recognition particles, which occur in 5% of caucasian adult PM/DM, 8% -13% of non-adult PM/DM and 2% JDM patients, and are found primarily in IMNM. The target antigen of the anti-HMGCR antibody is 3-hydroxy-3-methylglutaryl coenzyme A reductase, the generation of the antibody is related to the use of statins, and the antibody can be used as a diagnostic marker and a disease prognosis marker. The main clinical features of patients positive for anti-HMGCR antibodies are muscle weakness and dysphagia. The anti-cN 1A antibody was the first antibody found to be specific to sbm, and the target antigen was cytoplasmic 5' nucleotidase, whose relationship to clinical characteristics, response to therapy, and prognosis was to be studied further.
The presence of such autoantibodies is detectable in about 50% of the sera of adult PM/DM patients, but a significant proportion of patients have no autoantibodies detected in their sera. In clinical practice, autoantibody negative PM/DM patients do not receive a correct diagnosis and timely treatment. Because the pathogenesis and the etiology of PM/DM are not clear, and specific laboratory indexes for diagnosis are lacked, the diagnosis difficulty is high mainly according to muscle biopsy, electromyogram examination and clinical manifestation comprehensive judgment at present. Different types of idiopathic inflammatory myopathy can involve different target organs such as skin, lung and muscle, so the clinical manifestations are complex, early diagnosis and early treatment are difficult to achieve, and research reports that immune necrotic myopathy and muscular dystrophy patients have similar pathological changes, so part of patients are easy to misdiagnose. The early pathological changes of PM/DM generally belong to clinical treatment reversible transition or partial reversible transition, and the treatment effect is better; the later stage or the final stage belongs to the irreversible transition period of clinical treatment, and the disease is not good after healing, the death rate is high, and the consequence is serious. Therefore, early diagnosis and early treatment of PM/DM diseases are particularly important. The autoantibody can appear in a patient 10 years before the clinical symptoms of the autoimmune disease appear, even earlier, so that the practical early diagnosis and early effective treatment are thoroughly achieved, the serum antibody overall appearance of the PM/DM is further comprehensively and systematically researched, and a marker related to the PM/DM disease diagnosis is found, so that the autoantibody is the key point of the PM/DM disease research and the problem to be solved clinically urgently.
Disclosure of Invention
PM/DM is a rare group of autoimmune diseases with clinical heterogeneity. The autoantibody has high diagnostic specificity on PM/DM, and with the progress of detection methods, more and more autoantibodies related to PM/DM provide important reference significance for early detection, diagnosis, treatment and prognosis of diseases. However, the autoantibodies related to PM/DM reported at present are low in sensitivity, and the finding of a new marker is still a key point of research on PM/DM and a problem to be solved in clinic.
In order to solve the above problems, the signal peptide-recognizing particle 19 of the present invention, i.e., SRP19, or a fragment thereof is used for the preparation of an agent for diagnosing idiopathic inflammatory myopathy.
In a preferred embodiment of the present invention, the idiopathic inflammatory myopathy includes, but is not limited to, Polymyositis (PM), Dermatomyositis (DM), or polymyositis/dermatomyositis (PM/DM).
Wherein the diagnosing comprises: determining the level of antibodies reactive to SRP19 or a fragment thereof in a biological sample obtained from a patient exhibiting idiopathic inflammatory myopathy; optionally, the step of (a) is carried out,
comparing the level of antibodies in the biological sample to control data, wherein a detectable increase in antibodies reactive to SRP19 in the sample relative to the control data is indicative of a likelihood of developing idiopathic inflammatory myopathy.
Wherein the biological sample is a serum sample.
Wherein the level of anti-SRP 19 antibody is measured by the steps comprising:
a. contacting a biological sample from a patient with SRP19 or a fragment thereof;
b. forming an antibody-protein complex between an antibody present in the biological sample and SRP19 or a fragment thereof;
c. washing to remove any unbound antibody;
d. adding a detection antibody that is labeled and reactive with an antibody from the biological sample;
e. washing to remove any unbound labeled detection antibody; and
f. converting the label of the detection antibody to a detectable signal; wherein the presence of the detectable signal indicates the presence of anti-SRP 19 antibody in the patient.
Wherein said SRP19 or fragment thereof is deposited or immobilized on a solid support.
Wherein the support is in the form of a latex bead, a multi-well plate or a membrane strip.
Wherein the detection antibody is labeled by covalent attachment to an enzyme, a label with a fluorescent compound or metal, or a label with a chemiluminescent compound.
The present invention also provides a diagnostic kit for detecting and/or quantifying anti-SRP 19 antibodies in a biological sample, comprising: a solid support, wherein said SRP19 or fragment thereof is deposited on or immobilized on a solid support, and said anti-SRP 19 antibody is a biomarker for diagnosing idiopathic inflammatory myopathy.
Wherein the diagnostic kit further comprises a detection antibody that is labeled and reactive with an antibody from a biological sample.
Wherein the support is in the form of a latex bead, a multi-well plate or a membrane strip.
The invention adopts the high density protein chip technology to carry out PM/DM phase on 40 PM/DM patients, 30 autoimmune disease control patients and 20 healthy controlsAnd (3) screening target antigens of the autoantibodies, and performing statistical analysis on screening results to determine 91 PM/DM related autoantibodies. In order to confirm the sensitivity, specificity and clinical application value of the PM/DM related autoantibodies, a PM/DM related autoantibody target antigen chip is constructed and prepared by adopting 91 PM/DM related autoantibodies corresponding to target antigens, and clinical expansion sample verification is carried out on 397 cases of PM/DM patients, 200 cases of disease control patients (including 40 cases of RA, 40 cases of SLE, 40 cases of SS, 40 cases of SSc and 40 cases of chronic diseases) and 100 cases of serum of healthy controls. The results showed that the anti-SRP 19 antibody had 19.00% positive rate in the PM group, which was 2.67% higher than the positive rate in all the control groups (P ═ 3.26 × 10)-7) The difference in results was significant; the positive rate in DM group is 7.41%, which is higher than that in all control group by 2.67% (P ═ 0.80X 10)-2) The difference in results was significant; the positive rate in the PM/DM group is 10.33%, which is higher than that in all the control groups by 2.67% (P ═ 0.09X 10)-3) The difference in results was significant. Indicating that anti-SRP 19 antibodies are useful as diagnostic and detection markers for idiopathic inflammatory myopathy.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
437 PM/DM patients included in the invention were sourced from Beijing cooperative hospitals and hospitals subject support of industry funds. Of these, 131 male patients and 306 female patients were aged 2-79 years, with the mean age (44.07 ± 16.57) being the age of the patient. All PM/DM patients met the criteria for diagnosis of Bohan/Peter in 1975, and other autoimmune diseases such as Rheumatoid Arthritis (RA), Systemic Lupus Erythematosus (SLE), Sjogren's syndrome ((Sjogren's syndrome))
syndrome, SS), systemic sclerosis (SSc), and the like. 220 disease control groups, including 46 RA, group entry criteria: ACR differential diagnosis criteria in 1987; 47 SLE, entry criteria: the 1997 ACR triage diagnostic criteria; 47 SS, group entry criteria: AECG differential diagnostic criteria; 50 cases of SSc, grouping criteria: 2013 SSc classificationA diagnostic criteria; 40 other chronic disease controls. 120 healthy control groups were obtained from Beijing coordination hospital examination center. Strictly according to the research requirements of proteomics, samples are reserved, peripheral venous blood is collected, serum is separated within 2 hours and then subpackaged, and the blood is frozen at minus 80 ℃ for standby.
EXAMPLE 1 high Density protein chip screening for PM/DM related autoantibodies
All protein antigens on the high-density protein chip are provided with GST tags at the N ends for protein purification. First, the quality of the chip is verified by hybridizing the rabbit anti-GST monoclonal antibody with the chip protein. Then, 40 sera of PM/DM patients, 30 control sera of autoimmune patients and 20 healthy control sera were selected and hybridized with 90 high density protein chips to identify candidate PM/DM associated autoantigens through signal collection and data analysis.
1.1 extraction of high Density protein chip Scan data
Scanning a high-density protein chip by adopting a GenePix 4000B chip scanner, obtaining a TIFF format gray image, automatically completing the segmentation and extraction of each protein point signal pixel by analysis software according to an operation manual of GenePix Pro 6.0 software and sample point matrix parameters on the high-density protein chip, manually checking the signal intensity of all protein points on the chip one by one, manually adjusting the position and size deviation of the protein signal points caused by avoiding artificial factors such as impurities, scratches and the like, finally completing the extraction of all protein antigen point signal intensities on each high-density protein chip, generating a data document, and further analyzing and using the data document.
1.2 evaluation of the detection quality of the high-Density protein chip
After the gray image of the anti-GST antibody hybridized high-density protein chip is used, the Signal intensity of each protein point of the chip is collected, and when the Signal-to-Noise Ratio (SNR) of two parallel points of a probe on the chip is simultaneously greater than 2, the protein point on the chip can be detected. The ratio of protein spots detectable on the chip and the correlation of signal intensity between two parallel spots of each protein were then calculated.
1.3 high Density protein chip for detecting serum autoantibodies
The serum samples for the high density protein chip detection comprise 40 cases of PM/DM patients, 30 cases of autoimmune disease control patients and 20 cases of normal control. The specific operation steps are as follows:
(1) taking out the high-density protein chip stored at-80 ℃, isolating air to balance to room temperature, immersing the high-density protein chip in 5ml of protein chip sealing liquid containing 3% BSA PBST, and sealing for 1 hour (h) at room temperature;
(2) mu.l serum samples were taken according to 1:1000 of the serum is diluted in lml 3% BSA PBST, uniformly mixed by shaking, centrifuged at 10000rpm for 0min, and the supernatant is the diluted serum. And (3) after the protein chip is taken out from the sealing liquid, if the surface of the protein chip has air bubbles, sucking the sealing liquid in the incubation box by using an lml sample adding gun, washing the sealing liquid from the top end, sucking the redundant sealing liquid from one side by using absorbent paper to be dry as much as possible, and placing the sealing liquid in a wet box. Then sucking 150ul of diluted serum and adding the diluted serum onto a protein chip, slowly covering a cover glass to seal the chip without touching the chip so as to avoid damaging protein antigens on the chip, and incubating for 1h at room temperature;
(3) placing the incubated protein chip in 10ml of PBST lotion, carefully taking down the cover glass, placing the chip in a washing box, and rinsing for 10min each time for 3 times at room temperature PBST 40 rpm;
(4) alexa 647-labeled goat anti-human IgG antibodies were taken as follows: 1000 parts of the mixture is diluted in 3% BSA PBST, uniformly mixed by shaking, centrifuged at 10000rpm for 10min, and the supernatant is the diluted secondary antibody. The protein chip was removed from the washing box, and if there were air bubbles on the surface of the chip, the liquid in the incubation box was sucked up with a 1ml sample application gun and washed from the top, and the excess washing liquid was sucked up from one side with a water absorbent paper and placed in a wet box. Adding 150 mu l of diluted fluorescent secondary antibody to the chip, slowly adding a cover glass for sealing to avoid generating bubbles, and incubating the secondary antibody for 1h at room temperature in a dark place;
(5) placing the incubated protein chip in PBST lotion, carefully taking down the cover glass, placing the chip in a washing box, and washing for 10min each time for 3 times in dark at 40rpm of PBST;
(6) then distilled with double distilled water (ddH) at room temperature20) Rinsing the protein chip at 40rpm in dark for 3 times, each time for 10 min;
(7) taking out protein corePlacing the sheet in a 50ml centrifuge tube, centrifuging at 1000rpm for 2min with the side of the chip spotted with the antigen protein facing outwards, and spin-drying the residual ddH on the chip20；
(8) The GenePix 4000B chip scanner scans the protein chip, and the same scanning parameters are set for scanning the chip after each hybridization is completed.
1.4 determination of PM/DM related autoantibody target antigens
The signal intensity information of all protein spots on each high density protein chip collected by the GenePix Pro 6.0 software was imported into an Excel table. The foreground signal intensity (F635 mean) of each protein spot was divided by the background signal intensity (B635 mean) around the protein spot to serve as the signal value of the protein spot. I.e. Iij-F635 medium/B635 medium (Iij represents the signal value of protein i point in block j). The signal value of the protein antigen protein spot approaches 1, indicating that the lower the concentration of the corresponding autoantibody in the serum, the less easily the antibody is detected. Higher signal values indicate a greater ability of the autoantibody to bind to the target antigen protein in the serum sample. The average value of the signal intensity of two different spots of the same protein is taken as the signal intensity of the spot of the protein on the chip. When the average value is not less than 2, the protein spot is considered positive. Then, the information of the positive rate of the immunoreaction of each protein antigen on each serum and the high-density protein chip is counted. And performing data analysis on the PM/DM patient group, the autoimmune disease control group and the normal control group by adopting a common gene chip Significance Analysis (SAM) method in a high-throughput chip bioinformation analysis technology.
1.5 results
A total of 91 PM/DM associated target antigens were analytically determined and incorporated into the PM/DM associated autoantibody target antigen chiplet preparation, detailed in Table 1.
TABLE 1 proteins for preparing protein chips for PM/DM related autoantibodies target antigens
Example 2 construction of PM/DM related autoantibody target antigen protein chip and serum screening validation
The variety of autoantibodies produced by different patients with the same autoimmune disease is greatly different due to different genetic backgrounds and different environmental influence factors. In order to confirm whether the PM/DM related autoantibodies screened out by small samples by adopting a high-density protein chip have clinical significance and application value and the sensitivity and specificity of the autoantibodies to the diagnosis of PM/DM patients, the PM/DM related autoantibody target antigen screened out by the high-density protein chip in example 1 is prepared into a PM/DM related autoantibody target antigen protein chip for clinical amplification sample verification. Serum samples for clinical large sample verification detection of the PM/DM related autoantibody target antigen protein chip comprise 397 cases of PM/DM patients, 200 cases of disease control patients and 100 cases of healthy controls. The method comprises the following specific steps:
(1) taking out the PM/DM related autoantibody target antigen protein chip stored at-80 ℃, standing for 30min (avoiding condensed water), isolating air, recovering the room temperature, cutting off the package, carefully taking out the protein chip, facing to a fluorescent lamp, placing a fence on the front surface of the chip according to the position of each Block on the chip, pressing the fence tightly with force, and then placing the chip into a chip hybridization wet box with the front surface upward;
(2) add 50. mu.l of PBST (0.1% Tween20, v/v) buffer containing 3% BSA to each pen in a small square matrix and block for 1 hour at 37 ℃;
(3) placing the protein chip in a 50ml centrifuge tube, enabling the surface of the chip with the protein antigen to face outwards, and centrifuging at 1000rpm for 2min to remove liquid in each small square matrix;
(4) 1u1 serum was taken according to 1: diluting in lml 3% BSA PBST at a ratio of 1000, shaking and uniformly mixing, centrifuging at 10000rpm for 10min, and obtaining supernatant which is diluted serum;
(5) respectively adding 50ul of diluted serum into each small square matrix of the PM/DM related autoantibody target antigen protein chip to avoid generating bubbles, and incubating for 1h at room temperature;
(6) soaking the protein chip in 10ml of PBST washing liquor, and slowly shaking and rinsing the protein chip on a decoloring shaking table for 3 times, 40 revolutions per minute and 10min each time;
(7) taking out the protein chip from the washing box, placing the chip in a 50ml centrifuge tube, centrifuging for 2min at 1000rpm with the surface of the chip with the protein antigen facing outwards, and centrifuging to remove liquid in each small square matrix;
(8) alexa 647-labeled goat anti-human IgG antibodies were taken as follows: diluting with 1000% of BSAPBST, shaking and mixing uniformly, centrifuging at 10000rpm for 10min, and obtaining supernatant which is diluted secondary antibody;
(9) adding 50 microliter of A1: 1000 diluted A1exa647 labeled goat anti-human IgG antibody into each small matrix of the protein chip, and incubating for 1 hour at 37 ℃ in the dark;
(10) carefully tearing off the 14-hole fence adhered to the protein chip, soaking the chip in 10ml of PBST washing liquor, slowly shaking the chip on a decoloring shaking table, and rinsing the chip for 3 times in a dark place at 40rpm for 10min each time;
(11) then using 10ml ddH2The protein chip was rinsed 3 times for 10min each time in 0 dark. Placing the protein chip in a 50ml centrifuge tube, enabling the surface of the chip with the protein probe to face outwards, and centrifuging at 1000rpm for 2min to remove residual liquid on the chip;
(12) protein chip scanning was performed with a chip scanner. The parameters are set as follows: the excitation light wavelength was 635nm, the photomultiplier detector (PMT) was set to 650, the Power supply (Power) was set to 90, and the scanning pixels were set to 5 um.
The signal intensity information of all protein spots on each protein chip collected by the GenePix Pro 6.0 software was imported into an Excel table. The foreground signal intensity (F635 mean) of each protein spot was divided by the background signal intensity (B635 mean) around the protein spot to serve as the signal value of the protein spot. I.e., Iij-F635 medium-B635 medium (Iij represents the signal value of protein i point in block j). The closer the signal value Iij of the protein antigen protein spot approaches 0, the lower the corresponding level of autoantibodies in serum, the less easily the antibody is detected by the chip. A higher Iij signal value indicates a greater ability of the autoantibody to bind to the target antigen protein in the serum sample. The average value of the signal intensity of two different spots of the same protein is used as the signal intensity of the spot of the protein on the chip. The average signal intensities Iij (Averagenmamal) and Std of the target antigen were calculated for each protein spot of 100 healthy control groups included in the study, and Iij (Averagenmamal) +3Std was used as the positive response Cutoff value of the protein spot. Then, the positive rate information of each protein antigen immunoreaction on the chip of the serum and PM/DM related autoantibody target antigen protein of each research group is counted. The chi-square test was used to compare the positivity of autoantibody markers between different groups, and differences were considered significant when P < 0.01.
The results of the detection of 91 target antigens on the PM/DM related autoantibody target antigen chip are detailed in Table 2.
TABLE 2 detection results on a PM/DM related autoantibody target antigen chip
The results of the study showed that the anti-SRP 19 antibody had 19.00% positive rate in the PM group, which was 2.67% higher than the positive rate in all the control groups (P ═ 3.26 × 10)-7) The difference in results was significant; the positive rate in DM group is 7.41%, which is higher than that in all control group by 2.67% (P ═ 0.80X 10)-2) The difference in results was significant; the positive rate in the PM/DM group is 10.33%, which is higher than that in all the control groups by 2.67% (P ═ 0.09X 10)-3) The difference in results was significant. Indicating that anti-SRP 19 antibodies are useful as diagnostic and detection markers for idiopathic inflammatory myopathy.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the technical principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (5)
1. Use of signal peptide recognition particle 19, SRP19, in the manufacture of a reagent for the diagnosis of dermatomyositis, the diagnosis comprising: determining the level of antibodies reactive to SRP19 in a biological sample obtained from a patient exhibiting dermatomyositis; comparing the level of antibodies in the biological sample to control data, wherein a detectable increase in antibodies reactive to SRP19 in the sample relative to the control data is indicative of a likelihood of dermatomyositis, wherein the biological sample is a serum sample.
2. The use of claim 1, wherein the level of anti-SRP 19 antibody is measured by the steps comprising:
a. contacting a biological sample from a patient with SRP 19;
b. forming an antibody-protein complex between antibodies present in the biological sample and SRP 19;
c. washing to remove any unbound antibody;
d. adding a detection antibody that is labeled and reactive with an antibody from the biological sample;
e. washing to remove any unbound labeled detection antibody; and
f. converting the label of the detection antibody to a detectable signal; wherein the presence of the detectable signal indicates the presence of anti-SRP 19 antibody in the patient.
3. The use of claim 2, wherein said SRP19 is deposited or immobilized on a solid support.
4. Use according to claim 3, wherein the support is in the form of latex beads, a multi-well plate or a membrane strip.
5. The use of claim 2, wherein the detection antibody is labeled by covalent attachment to an enzyme, a label with a fluorescent compound or metal, or a label with a chemiluminescent compound.
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| CN102138072A (en) * | 2008-09-01 | 2011-07-27 | 学校法人庆应义塾 | Diagnosis method and diagnosis kit for dermatomyositis |
| CN103154736A (en) * | 2010-05-13 | 2013-06-12 | 新泽西医科和牙科大学 | Diagnostic autoantibody profiles for the detection and diagnosis of neurodegenerative diseases |
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| CN102138072A (en) * | 2008-09-01 | 2011-07-27 | 学校法人庆应义塾 | Diagnosis method and diagnosis kit for dermatomyositis |
| CN103154736A (en) * | 2010-05-13 | 2013-06-12 | 新泽西医科和牙科大学 | Diagnostic autoantibody profiles for the detection and diagnosis of neurodegenerative diseases |
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| Title |
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| Deternimation of Anti-SRP Antibodies;Prof.R.L.Humbel;《DIARECT Newsletter》;20171231;第1-6页 * |
| Expression of anti-SRP19 antibody in muscle tissues from patients with autoimmune necrotizing myopathy;Q. Wang等;《Genetics and Molecular Research》;20161231;第15卷(第3期);第1-8页 * |
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