CN101329302A - Protein fingerprint pattern model for systemic lupus erythematosus diagnosis - Google Patents
Protein fingerprint pattern model for systemic lupus erythematosus diagnosis Download PDFInfo
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Abstract
The invention discloses a protein fingerprint atlas model used for the diagnosis of systemic lupus erythematosus. The invention adopts a new technical platform (Clinprot) which uses sample processing and substrate auxiliary laser analysis ionization flying time mass spectrometry on the basis of reverse magnetic beads; analysis is carried out by using the software Clinprotools Software 2.1 after the data is obtained; the serum protein fingerprint atlas of the sample is established, analyzed and screened so as to obtain difference polypeptide; the difference polypeptide is used for establishing and verifying the protein fingerprint atlas model with high sensitiveness and high specificity corresponding to the disease. The protein fingerprint atlas model can be used for the diagnosis of the systemic lupus erythematosus, has important significance for the clinical diagnosis and disease understanding of the systemic lupus erythematosus, and provides the new concept and new basis for researching the pathogenesis and the early diagnosis of the SLE from the point of view of proteomics.
Description
Technical field
The present invention relates to the medical diagnostic techniqu field, relate in particular to a kind of protein fingerprint pattern model that is used for systemic lupus erythematosus diagnosis.
Background technology
Systemic loupus erythematosus (systemic lupus erythematosus, SLE) be the autoimmune disease of involving a plurality of systems of whole body, after glucocorticoid and immunodepressant are used to treat SLE, though patient's prognosis has clear improvement, but generally speaking should disease treatment present situation still not allow undue optimism, 15% the patient of still having an appointment is dead or renal failure occurs in morbidity 5 years, and recurrence also is a severe problem that is faced with in the therapeutic process simultaneously.SLE is the prototype of autoimmune disease, can involve multiple internal organs or system clinically.The SLE tissue damage mainly is owing to immune complex deposit causes in corresponding tissue.The clinical disease spectrum is very wide, has to alleviate and worsen the feature that alternately takes place.In order better to treat and control SLE, be necessary in time and effectively to judge disease activity and the degree of impairment of SLE.
Need to seek a kind of biomarker that can react disease active situation at that time, this mark also should have disease specific, to the variation sensitivity after the treatment, can predict lapsing to of disease.Single mark may not satisfy all these requirements, therefore need unite and use the unlike signal thing to be used for the different molecular incident of monitoring of diseases generating process.Have thousands of peptide species in the human serum, the overwhelming majority in the middle of them is considered to partly be decomposed by the endogenous protein digestive enzyme fragment of larger molecular weight protein gained, and the definite character of these polypeptide does not also have resolved.But, studies show that complex set (peptide group) that these polypeptide form can judge the biology incident of whole machine body as a kind of novelty and ripe Forecasting Methodology.
In case after the lupus erythematosus clinical symptoms occurs, SLE is made rapid diagnosis and suitably treats the huge challenge that remains the physician.Therefore, need to seek new more hypersensitivity and the specific lupus diagnosis marker of having.
Summary of the invention
The object of the present invention is to provide a kind of protein fingerprint pattern model that is used for systemic lupus erythematosus diagnosis, it can high sensitivity, specially property detects systemic loupus erythematosus.
The present invention adopts a kind of new technology platform (Clinprot), and this platform has used based on the sample preparation of reverse magnetic bead and ground substance assistant laser desorption ionization flight time mass spectrum (MALDI-TOF MS) and analyzed.At first systemic loupus erythematosus (SLE) patient is divided into two groups of active stage and stationary phases according to the SLEDAI-2000 standards of grading, and choose rheumatoid arthritis (RA) disease control group and normal healthy controls group, collect blood, prepare serum, with reverse magnetic bead is that sample preparation is carried out on the basis, carrying out MALDI-TOF MS with ground substance assistant laser desorption ionization flying time mass spectrum analysis instrument detects, obtain analyzing with Clinprotools Software2.1 software after the data, set up the serum protein fingerprint of four groups of samples, analyze and screen the otherness polypeptide, the utilization variance polypeptide is set up and the hypersensitivity of checking disease association and the protein fingerprint pattern model of high specific.Mass-to-charge ratio m/z and protein peak strength factor Arb.U thereof according to each protein have set up the protein fingerprint pattern model of differentiating between systemic loupus erythematosus, rheumatoid arthritis and normal person, said specific protein mass-to-charge ratio m/z and peak intensity coefficient Arb.U are respectively m/z=3192.34, Arb.U≤8.47; M/z=9289.29, Arb.U≤15.56; M/z=4267.96, Arb.U≤1.91; M/z=2952.17, Arb.U≤13.47; M/z=2660.7, Arb.U 〉=33.04; M/z=3263.03, Arb.U≤15.52; M/z=1778.75, Arb.U 〉=1.93; M/z=5904.74, Arb.U≤66.26; Wherein the protein fingerprint pattern model differentiated of Patients with SLE and normal person is by m/z=3192.34, Arb.U≤8.47; M/z=9289.29, Arb.U≤15.56; M/z=4267.96, Arb.U≤1.91; M/z=2952.17, Arb.U≤13.47 are drawn and are formed; The protein fingerprint pattern model that patient with rheumatoid arthritis and normal person differentiate is by m/z=2660.7, Arb.U 〉=33.04; M/z=3263.03, Arb.U≤15.52; M/z=1778.75, Arb.U 〉=1.93; M/z=5904.74, Arb.U≤66.26 are drawn and are formed.
Described software is Clinprotools Software2.1.
Its identification system lupus erythematosus and normal person's discrimination is 92.36%, and predictive ability is 90.27%; Its discrimination of differentiating rheumatoid arthritis and normal person is 100%, and predictive ability is 87.88%.
Utilize this protein fingerprint pattern model, can be used for systemic lupus erythematosus diagnosis, clinical diagnosis and state of an illness understanding for systemic loupus erythematosus have great importance, and provide new approaches and new foundation for inquire into the pathogenesis of SLE and diagnosis in early days from the angle of proteomics.
The Clinprot technology platform that the present invention adopts has used based on the sample preparation of reverse magnetic bead and MALDI-TOF MS and has analyzed, the magnetic capture analysans is caught sensitiveer than chip surface, because spherical particle has bigger mating surface, therefore bigger than the chip of minor diameter in conjunction with volume.It can detect a large amount of polypeptide in the serum simultaneously.This technology platform has hypersensitivity and resolution, it can detect 400 multiple polypeptides that surpass that molecular weight ranges is 800-15000Da in one bleeds clearly, the protein fingerprint pattern model of setting up with the method that is used for systemic lupus erythematosus diagnosis has high sensitivity and property specially.The application of automatic fluid processing machine arm has guaranteed the high flux and the repeatability of this technology.
Description of drawings
Fig. 1 is the technology of the present invention route map.
Embodiment
1 material
1.1 key instrument
" red lid " vacuum test tube Guangzhou Yangpu Medical Articles Co., Ltd.
Z4M bar code printer Zebra technology company
Genesis Freedom 100 liquid machine arm Tecan companies
Robotization rifle head Tecan company
Long magnet (2.75 * 0.25 * 0.125 inch) K﹠amp; D magnetic material company
Disk magnet (0.25 inch of diameter, 0.25 inch of thickness) Forcefield company
Customization packing pipe support
Customization side direction magnet stand
Customization bottom magnet stand
The non-shirt rim of template III PCR plate USA scientific company
Template III PCR overskirt side plate USA scientific company
Oscillator Barnstead/Thermolyne company
PCR refrigeratory initiating means USA scientific company
Autoflex MALDI-TOF mass spectrometer Bruker company
Bruker 384-point MALDI target plate Bruker company
Gemini software Tecan company
FlexControl software Bruker company
FlexAnalysis software Bruker company
1.2 main agents
A-cyanogen-4-hydroxy-cinnamic acid Agilent company
Mark Bruker company in mark and the albumen in the polypeptide
Artificial synthetic polypeptide 3897 Sigma companies
Cromoci Sigma company
Dynabeads WCX Dynal company
Second cyanogen Burdick﹠amp; Jachkson company
Trifluoroacetic acid (TFA) Piere company
Human serum Sigma company
Milli-Q water system USFilter company
10 * PBS Bio-Rad company
Isopropyl alcohol J.T.Baker company
1.3 sample
(Systemic Lupus Erythematosus, SLE) patient's 50 examples are the women to systemic loupus erythematosus, at 14～54 years old age (The median age 35 years old), all meet the diagnostic criteria that American Rheumatism Association (ARA) revises nineteen eighty-two.According to the SLEDAI-2000 standards of grading patient SLE is divided into two groups of active stage (SLEDAI＞8) and stationary phases (SLEDAI≤8), all cases were all got blood before increasing the glucocorticoid consumption or adding with endoxan.In addition, (Rheumatoid Arthritis, RA) patient all meet the diagnostic criteria of American Rheumatism Association 1987 revision, select 24 of sex and agematched healthy volunteers as control group to select 25 routine rheumatoid arthritiss.
2 operation stepss
2.1 collect blood and prepare serum
Collect venous blood to vacuum test tube.
Slowly put upside down heparin tube 5 times, set accelerator is fully mixed with blood, the vertical down placement heparin tube of room temperature condition made blood clotting in 1 hour.
Place heparin tube on ice, carry out next step in 2 hours.
Under the room temperature condition, 2000g, centrifugal 10 minutes.
The EP pipe sticks the label corresponding with sample.
With pipettor upper serum is transferred to the EP pipe that posts label, every pipe 1ml, this EP pipe that installs serum is exactly " blood serum sample comes source capsule ".
All blood serum samples are kept at-80 ℃ immediately, avoid multigelation.Blood serum sample need be placed and be transported to the mass spectrophotometry laboratory on the dry ice.
2.2 for blood serum sample comes the source capsule coding
All samples information is input to Laboratory Information Management System.When submitting sample message to, the bar coded sticker of primary sample pipe will generate automatically at every turn.Sort so that subsequent treatment is easier sample is the same with label.Sample hose is labelled to be generated and confirmatory sample pipe inventory simultaneously giving.
Need sample hose is positioned on the dry ice when labelling to sample hose, preceding sample hose need being dried with assurance with paper handkerchief of labelling is adjacent to.
Install the sample hose that posts label with box,, be positioned over-80 ℃ of refrigerators then at the good problem title of box foreign labeling, date and " sample hose to be packed " printed words.
2.3 packing sample
The box that sample hose to be packed is housed is taken out refrigerator, place sample hose on dry ice.
Scan the bar code on first sample hose to be packed.If database can correctly be discerned sample, one " sample identity " will generate so.Select serum volume and every pipe volume to be packed in the sample hose, calculate branch tubulature number, print the new label that all divide tubulature.
The packing bar coded sticker is attached on the clean 0.5mlEP pipe, arranges according to the order of primary sample pipe and divide tubulature.The branch that these minutes tubulature directly is put into the fluid operated device of Tecan is shelved, and lines up 16 row, every row 9 pipes.
All branch tubulatures post label and arrange and confirm good after, with sample hose be positioned over wet on ice, will be placed on the big ice chest by the order of packing according to them and melt 60min.
The branch of aluminum shelved be placed on the automatic machinery carrier, sample hose also places on the frame according to the order of minute tubulature.Should be positioned over the sample pipe support before dividing process of assembling to begin and wet on ice.
Open the Gemini program on the desktop computer, operation Gemini packing Automatic Program is carried out the branch process of assembling.
Cover the branch tubulature, they are placed on the dry ice, each sample is only got tubulature in a fen and is done next step analysis, and remaining is placed on-80 ℃ of refrigerators.
Be placed on tubulature in to be analyzed minute in the box separately and post label.
2.4 blood serum sample randomization ordering
Scan the bar code of tubulature in to be analyzed minute, set up an inventory, preserve this inventory with the common text documents form.Divide tubulature to be positioned on the dry ice always.
Operation randomization software is imported the number of sample to be analyzed, and the once multipotency of each MALDI target is analyzed 96 parts of packing serum, if added more parts serum, program will generate two or more MALDI target plate and sort.
Press START button, program can require select File (that common text documents of just having set up), in case after you selected this file, the stochastic distribution position of sample on the MALDI target promptly can show, in addition, also have interior target position also can show.The position of blood serum sample before doing magnetic bead sorting in 96 orifice plates do not represented in the ordering that randomization software generates.
Subsequently, program can be inquired the catalogue of preserving output file (implementing file automatically), and output file includes the instruction that makes the automatic mass spectrophotometry of FlexControl program run.
Need to print these targets figure, because they will need in the randomization ordering of packing serum.
According to the order that shows among the target figure, packing serum is placed on 96 orifice plates of precooling.Constantly want check sample inventory and target figure to confirm that sample, information among the target figure and the sign of branch tubulature in the inventory are consistent.Divide tubulature to be positioned on the dry ice always.
Output file is transferred in the MALDI-TOF computer.
2.5 set up the robotization handling procedure
At first, need to confirm following several: all randomized 96 orifice plates that are positioned over of all testing samples, check 2 times to confirm each sample all in the tram, then they are put into-80 ℃ of refrigerators; Reagent that all are required and material are all ready.
The foundation of robotization handling procedure is chosen in to be carried out morning, cleans Tecan liquid machine arm, bleeds to the water that machine will use, and guarantees the Litter that refuse container has enough big space to accept to produce in the analytic process, confirms that automatic suction nozzle is clean.Twice of updating system.
Carry out Quality Control test (specifically seeing for details hereinafter).
Begin the sample that thaws, place wet 60min on ice, guarantee that sample puts order in plastic processing frame.
The magnetic bead of 320 μ l uses 320 μ l deionized waters resuspended with 200 each washed twice of μ l deionized water then, to remove the ethanol in the magnetic bead storage liquid.
The about 5min of mixing magnetic bead soft in oscillator thoroughly disperses up to magnetic bead, and magnetic bead is drawn onto 0.2ml thin-walled octal bar, and every hole 40 μ l are put into the appropriate location of machine deck then.
TFA with freshly prepared 0.1% joins the TFA groove.
Begin to prepare interior mark, see table 1 for details, interior mark must preparation on the same day.
Prepare one the 96 half of skirtboard in hole, the every hole of first row adds the second cyanogen of 75 μ l 50%, and the every hole of secondary series adds the matrix solution of 95 μ l.The 3rd row A round adds mark in the 65 μ l " master mix ", and the 3rd row B round adds 70 μ l matrix solutions.
The half of skirtboard in top preparation 96 holes is placed into cooling frame and fastens, cooling frame needs could use at-20 ℃ of precooling several hrs.
The MALDI target is put into the precalculated position.
Mark in the table 1
a?All?the?m/z?are?calculated?for?single-charged?ions?except?when?otherwise?it?is?indicated.
b?double-charqed?ion.
2.6 half sample segment is handled
Each sample 50 μ l is joined the appropriate location in 48 holes, left side of no shirt rim 96 orifice plates, and the corresponding randomization of sample for reference position is to confirm.This 96 orifice plate is placed into the relevant position of machine deck.
Whether updating system is once checked ready again.
Operation Tecan program is to carry out following operation:
Blow and beat the magnetic bead in the mixing octal bar repeatedly 10 times with rifle.
Each automatic gun head is drawn the magnetic bead in the 40 μ l octal bars.
Abandon 5 μ l to avoid when magnetic bead mixes with serum, containing bubble in the rifle head.
In containing 48 holes of sample, every hole adds 5 μ l magnetic beads.
Slowly blow and beat 5 times to mix magnetic bead and serum.
96 orifice plates are transferred to the side direction magnet stand so that magnetic bead is adsorbed onto sidewall.
Siphon away and abandon supernatant.
The TFA solution that adds 200 μ l 0.1% in every sample well.
Move forward and backward 96 orifice plates, the washing magnetic bead.
Siphon away and abandon washing lotion.
Repeated washing once.
After the washing, slowly draw and abandon 80 μ l washing lotions for the second time.
Plate is transferred to original position.
Blow and beat up and down 5 times with rifle, magnetic bead is resuspended in the remaining washing lotion.
96 orifice plates are transferred on the magnet stand of bottom, waited for 160s, make magnetic bead be adsorbed to the pipe end.
Siphon away and abandon remaining washing lotion very carefully.
The second cyanogen that adds 6 μ l 50% in every sample well.
Blow and beat 10 times fast up and down with resuspended magnetic bead.
Plate is transferred to the side direction magnet stand.
Shift in every sample well 5 μ l eluents in 48 holes on 96 orifice plate right sides.
In 48 holes on right side, add the prefabricated matrix solution of 10 μ l.
Piping and druming mixed-matrix and eluent.
Draw 1 μ l mixed liquor and with its point to the MALDI target.
Interior mark liquid in the microwell plate that will place on cooling frame and matrix liquid mix at 1: 1, blow and beat up and down 5 times.
Draw 90 μ l potpourris, in every hole of the A of another non-shirt rim 96 orifice plates row, add 10 μ l.
Draw 8 μ l, add 1 μ l at the interior punctuate of each MALDI target.Though the potpourri in each on the punctuate is identical, their position differences on target.
2.7 latter half sample preparation
Prepare 48 parts of serum that next group is analyzed.Begin the sample that thaws, place wet 60min on ice, guarantee that sample puts order in plastic processing frame.
Serum is added new no shirt rim 96 orifice plates, place and wet on ice.
Remove and abandon the microwell plate that has serum and magnetic bead after first analysis finishes from the side direction magnet stand.Remaining step is identical with the process of first sample analysis.
Open the Gemini program, select the point sample order of the second half parts of 384 MALDI targets.After checking mistake, updating system twice, working procedure, program will be carried out identical step, unique different be that the potpourri of eluate and matrix is specifically by o'clock in the second half parts of 384 o'clock MALDI targets.
Remove the MALDI target plate, it is transferred to the MALDI-TOF mass spectrometer, target plate must be read the result in 3-4 hour.
2.8 mass spectrophotometry and collection of illustrative plates output
Target plate to be analyzed is inserted into the MALDI-TOF mass spectrometer.
Open the correct file that automatically performs, beginning is operation following steps (" automatic execution method " must upgrade could use suitable Laser Power Supply) automatically:
Transfer to first target spot of MALDI plate.
Begin to collect the peptide spectrum of fragment in the 0.7-4kDa m/z scope: 400 laser of average emitted, divide four groups to aim at four different positions, matrix point surface, to launch 100 times for every group, frequency is 50Hz.Collection of illustrative plates obtains under the geometric state of linear model, and ion quickens (the time-delay extraction voltage is 18.6kv) under the 20kv, and multiplier tube voltage is-1.3kv that it is m/s=400 that the ion grid are set.The time-delay time of drawing remains on 80ns, has reasonable time to postpone energy focusing behind each emission laser like this.The effective laser energy that is sent on the target is controlled at each emission 16 μ J (± 10%) (down together).Whole illumination procedure is controlled according to following steps automatically by " operation automatically " function of instrument: be provided with at the instrument of optimizing and adopt correct collection of illustrative plates acquisition method down, find the correct target spot on the MALDI plate, carry 4 groups of (100 times every group) Laser emission (organizing to next) from one group of helical motion.
Move to next target spot, gather the peptide spectrum in the 0.7-4kDa m/z scope.Continue to gather the collection of illustrative plates in all (comprising sample and interior mark) these mass ranges then.
Transfer to first target spot of MALDI plate once more.Begin to collect first peptide spectrum of fragment in the 4-15kDa m/z scope: 500 laser of average emitted, divide five groups to aim at five different positions, matrix point surface, to launch 100 times for every group, frequency is 50Hz.Collection of illustrative plates obtains under the geometric state of linear model, and ion quickens (the time-delay extraction voltage is 18.6kv) under the 20kv, and multiplier tube voltage is-1.3kv that it is m/s=3000 that the ion grid are set.The time-delay time of drawing remains on 50ns, has reasonable time to postpone energy focusing behind each emission laser like this.Can be sent to initial 100 identical zones of Laser emission with first mass range (4-15kDa) in initial 100 Laser emission of second mass range (4-15kDa), because for each MALDI target spot, AutoPlay function can be remembered an initial position.Therefore, need 100 times extra Laser emission in second mass range (4-15kDa), reason is that initial position does not exist crystal.
Move to next target spot, gather the peptide spectrum in the 4-15kDa m/z scope.Continue to gather the collection of illustrative plates in all (comprising sample and interior mark) these mass ranges then.
Open the FlexAnalysis program after analyzing end, use " opening a plurality of collection of illustrative plates " function to open all mass spectrograms that just have been established, check whether all mass spectrums generate.
" processing " function by FlexAnalysis is carried out atlas analysis.In addition, also collection of illustrative plates can be converted to the text document that label is separated, like this, each collection of illustrative plates contains x and two variablees of y.
2.9 quality control
Carry out a quality control testing weekly with assessment Tecan liquid machine arm and the mass spectrometric repeatability of AutoFlexMALDI-TOF.The human serum that uses market to buy, each Quality Control Analysis gets final product with 2 pipes, guarantees that there are enough samples in 10 holes in 96 orifice plates, the PBS filling of remaining hole.The distribution of blood serum sample on microwell plate will all even dispersion.The Quality Control analytical procedure is as follows:
At the wet serum 1 hour of thawing on ice.
Simultaneously, prepare Tecan self-reacting device and test agents useful for same.
Operation Gemini program (identical) with the clinical sample test.
Remove target plate, transfer to the MALDI-TOF mass spectrometer.
Insert target plate to be analyzed, system of selection file " serum_1-4kda.par ".
Check and write down the state of MS laser, open " state tag ", click " in detail ", check that the state of laser is also adjusted laser energy to 16 μ J.
Open quality control and implement template automatically, edit this file, change the destination folder of date and preservation collection of illustrative plates.Behind the editor, be kept at quality control and implement file directory automatically.
Automatic enforcement file and bootup window after in the FlexControl program, opening editor.
Open the FlexAnalysis program after operation finishes, use " opening a plurality of collection of illustrative plates " function to open all Quality Control mass spectrograms that just have been established.
Stack this 10 mass spectrograms, in addition, amplify 942.43,1,211.70,1,449.76 and 1,864.95 this zone, four peptide peaks respectively, measure the intensity at these four peaks in the 10 secondary collection of illustrative plates respectively.The variation of the mean intensity at each peak should be not more than 20-30%.
2.10 statistical study
Clinprotools Software2.1 is used in data analysis work.Statistical test method among the Clinprotools (parameter T-Test and nonparametric technique Wilcoxon Test) is sought differential protein, analyze the polypeptide of variant trend, and utilize genetic algorithm in the software in conjunction with KNN (k-nearest neighboure, k=1,3,5,7) set up the classification forecast model.At first use genetic algorithm, establishing aberration rate is 0.2, and crossing-over rate is 0.5, and the initial chromosome number is 1000, fitness function is finally evolved through 10000 times with the accuracy rate of KNN result of determination, traversal k, finally in differential protein, select wherein otherness polypeptide for use, set up model.The specificity of computation model, susceptibility and average accuracy rate.With arbitrary sampling method (select 80% sample to set up model at random, remaining 20% conduct checking sample moves ten times), the validity of verification model.
Embodiment 1 identification system lupus erythematosus patient and healthy people
Collect venous blood and prepare serum, obtain its serum protein fingerprint according to above-mentioned steps, the protein fingerprint pattern model that Patients with SLE and normal person differentiate, if person under inspection m/z=3192.34, Arb.U≤8.47; And m/z=9289.29, Arb.U≤15.56; And m/z=4267.96, Arb.U≤1.91; And m/z=2952.17, Arb.U≤13.47 are diagnosed as Patients with SLE.
Embodiment 2 differentiates patient with rheumatoid arthritis and healthy people
Collect venous blood and prepare serum, obtain its serum protein fingerprint according to above-mentioned steps, the protein fingerprint pattern that patient with rheumatoid arthritis and normal person differentiate, if person under inspection m/z=2660.7, Arb.U 〉=33.04; And m/z=3263.03, Arb.U≤15.52; And m/z=1778.75, Arb.U 〉=1.93; And m/z=5904.74, Arb.U≤66.26 are diagnosed as patient with rheumatoid arthritis.
Claims (3)
1, the protein fingerprint pattern model that is used for systemic lupus erythematosus diagnosis, it is characterized in that: the mass-to-charge ratio m/z of a plurality of specific proteins of Patients with SLE that the serum proteins wave spectrum process software statistics analysis that is detected by ground substance assistant laser desorption ionization flying time mass spectrum analysis instrument system obtains and protein peak strength factor Arb.U thereof draw and form, said specific protein mass-to-charge ratio m/z and peak intensity coefficient Arb.U are respectively m/z=3192.34, Arb.U≤8.47; M/z=9289.29, Arb.U≤15.56; M/z=4267.96, Arb.U≤1.91; M/z=2952.17, Arb.U≤13.47; M/z=2660.7, Arb.U 〉=33.04; M/z=3263.03, Arb.U≤15.52; M/z=1778.75, Arb.U 〉=1.93; M/z=5904.74, Arb.U≤66.26; Wherein the protein fingerprint pattern model differentiated of Patients with SLE and normal person is by m/z=3192.34, Arb.U≤8.47; M/z=9289.29, Arb.U≤15.56; M/z=4267.96, Arb.U≤1.91; M/z=2952.17, Arb.U≤13.47 are drawn and are formed; The protein fingerprint pattern model that patient with rheumatoid arthritis and normal person differentiate is by m/z=2660.7, Arb.U 〉=33.04; M/z=3263.03, Arb.U≤15.52; M/z=1778.75, Arb.U 〉=1.93; M/z=5904.74, Arb.U≤66.26 are drawn and are formed.
2, the protein fingerprint pattern model that is used for systemic lupus erythematosus diagnosis according to claim 1 is characterized in that: described software is Clinprotools Software2.1.
3, be used for the method for building up of the protein fingerprint pattern model of systemic lupus erythematosus diagnosis, may further comprise the steps:
1) selecting system lupus erythematosus group, rheumatoid arthritis disease control group and normal healthy controls group are collected blood, are prepared serum;
2) be that sample preparation is carried out on the basis with reverse magnetic bead;
3) carrying out ground substance assistant laser desorption ionization flight time mass spectrum with ground substance assistant laser desorption ionization flying time mass spectrum analysis instrument detects;
4) obtain analyzing with Clinprotools Software2.1 software after the data, set up the serum protein fingerprint of sample, analyze and screen the otherness polypeptide, the utilization variance polypeptide is set up and the hypersensitivity of checking disease association and the protein fingerprint pattern model of high specific.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101916325A (en) * | 2010-06-30 | 2010-12-15 | 戴勇 | Stage protein expression difference spectrum model for systemic lupus erythematosus and construction method |
| CN101916325B (en) * | 2010-06-30 | 2013-08-28 | 戴勇 | Stage protein expression difference spectrum model for systemic lupus erythematosus and construction method |
| CN102262097A (en) * | 2011-04-27 | 2011-11-30 | 戴勇 | SLE (systemic lupus erythematosus) spectrogram model construction method and SLE spectrogram model |
| CN102262097B (en) * | 2011-04-27 | 2014-07-02 | 戴勇 | SLE (systemic lupus erythematosus) spectrogram model construction method and SLE spectrogram model |
| CN102818837A (en) * | 2012-04-17 | 2012-12-12 | 深圳出入境检验检疫局动植物检验检疫技术中心 | Protein fingerprint model, system and application thereof for anthrax |
| CN105548338A (en) * | 2015-12-10 | 2016-05-04 | 山东出入境检验检疫局检验检疫技术中心 | Protein fingerprint model for Burkholderia gladioli and application thereof |
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