CN107142325A - A kind of gene marker and its application for medicine anaphylactoid reaction sensitive group examination - Google Patents
A kind of gene marker and its application for medicine anaphylactoid reaction sensitive group examination Download PDFInfo
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- CN107142325A CN107142325A CN201710509594.2A CN201710509594A CN107142325A CN 107142325 A CN107142325 A CN 107142325A CN 201710509594 A CN201710509594 A CN 201710509594A CN 107142325 A CN107142325 A CN 107142325A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
Abstract
The invention discloses a kind of gene marker for medicine anaphylactoid reaction sensitive group examination and its application, the gene marker includes peripheral blood MrgprX2 characterizing gene sequences.The invention also discloses application of the gene marker in the product for preparing examination early stage medicine anaphylactoid reaction sensitive group.The gene marker proposed by the present invention, it can be used in the examination of medicine anaphylactoid reaction sensitive group, there is sensitiveness height, high specificity, and due to being to be used as detection sample using the peripheral blood clinically most easily gathered during examination, it is easy to detect, suitable for the large-scale crowd examination and diagnosis of medicine anaphylactoid reaction sensitive group, have a extensive future.
Description
Technical field
Sieved the present invention relates to technical field of molecular biology, more particularly to a kind of medicine anaphylactoid reaction sensitive group that is used for
The gene marker looked into and its application.
Background technology
Medicine, which causes allergic reaction, is broadly divided into I types (anaphylactic type), II types (cell toxicant type), type III (immune complex
Type), (delayed) allergic reaction of IV types and anaphylactoid reaction.The clinical symptoms of anaphylactoid reaction are similar with allergic reaction.But its
Reaction mechanism and type i allergic reaction are simultaneously differed.After medicine enters in vivo, mast cell release content can be directly resulted in,
Allergic reaction is triggered, and clinical symptoms are serious.Anaphylactic shock betides the individual that only a few receives therapeutic dose medicine, does not have
Sensitization process, patients serum's Ig E concentration also has no rise, but shows as typical anaphylactic type symptom, therefore quilt again
Referred to as false drug allergy.Anaphylactoid reaction is unrelated with antigen specific immune reaction, is sent out in such medicine of first contacts
It is raw, and have certain correlation with dosage, injection speed., not only can be special as the mast cell of allergy main effects device
Property IgE attachments and activating causes degranulation, cause I type allergy, can also be connect by a variety of secretagogues such as inflammatory factor, medicine are upright
Activation causes cell degranulation, causes clinical common anaphylactoid reaction, patient skin damaged symptom is occurred, that is, show erythema,
Nettle rash, oedema etc., severe patient can cause the symptoms such as asthma, shock.The mechanism that current anaphylactoid reaction is produced not yet is recognized completely
Know, histamine release of mast cell may be directly stimulated with sensitizer, or directly by the anaphylatoxin such as C5a, C3a, C4a and hypertrophy
The mast cell degranulation that cell-specific membrane receptor is mediated after combining is relevant.
The clinical manifestation of anaphylactoid reaction is local skin symptom, such as blood vessel dilatation of Head And Face, chest and four limbs, red
Spot, oedema, conjunctival congestion；Gastrointestinal symptom, such as salivation, nausea,vomiting,diarrhea, apocleisis；Respiratory circulatory system symptom,
Such as uncomfortable in chest, palpitaition, blood pressure rise, expiratory dyspnea, serious occurs respiratory and circulatory failure, causes patient's shock, death.Its disease
Shape feature is relevant with the speed that medical intravenous is injected, and injection speed is faster, and the burst size of histamine is bigger, and body reaction is more serious,
That is low dosage, low concentration, can without or only there are light symptoms；Heavy dose of, high concentration can induce serious adverse drug reaction.But
It is that the reaction symptom of body can gradually weaken with repeat administration or even voluntarily disappear, and allergic reaction does not have this feature then.
Occurrence frequency of the anaphylactoid reaction in recent years in clinic is constantly raised, and accounts for the 77% of acute allergic reaction,
In anaphylactic shock caused by traditional Chinese medicine, about 3/4 belongs to anaphylactoid reaction, because its clinical harmfulness is big, is increasingly becoming
The focus of medical worker concern.But, do not monitor the respective standard of anaphylactoid reaction clinically at present, more do not shift to an earlier date
Detect that the correlative study of anaphylactoid reaction risk occurs for patient.Conventional mark histamine, trypsinlike enzyme and IgE exists each
The problem of planting various kinds, is not particularly suited for clinical detection anaphylactoid reaction.
The content of the invention
It is an object of the invention to provide a kind of gene marker for medicine anaphylactoid reaction sensitive group examination and
It is applied, and occurs the problem of anaphylactoid reaction Risk Screening technological means is short of to solve current patient.
To reach above-mentioned purpose, the present invention is realized using following technical scheme：
A kind of gene marker for medicine anaphylactoid reaction sensitive group examination, the gene marker is
The characterizing gene sequence of MrgprX2 acceptors.
The characterizing gene sequence of the MrgprX2 acceptors is as shown in SEQ ID NO.1.
The described gene marker for medicine anaphylactoid reaction sensitive group examination is preparing medicine anaphylactoid reaction
Application in sensitive group examination product.
The medicine anaphylactoid reaction sensitive group examination product includes using real time quantitative PCR method examination medicine class allergy
React the product of sensitive group.
The product of the use real time quantitative PCR method examination medicine anaphylactoid reaction sensitive group includes specific amplification
The primer of the characterizing gene sequence of MrgprX2 acceptors.
The primer includes Forward primers and Reverse primers, the wherein sequence of Forward primers such as SEQ ID
Shown in NO.2, the sequence of Reverse primers is as shown in SEQ ID NO.3.
Compared with prior art, the invention has the advantages that：
Gene marker provided by the present invention for medicine anaphylactoid reaction sensitive group examination is MrgprX2 acceptors
Characterizing gene sequence.The gene marker can be used in the examination of medicine anaphylactoid reaction sensitive group, can prepare medicine
It is applied in terms of anaphylactoid reaction sensitive group examination product, the gene marker occurs available for patient's anaphylactoid reaction
The examination and diagnosis of risk, with very high Sensitivity and Specificity, and due to using clinically most easy during examination
The peripheral blood of collection is detected that MrgprX2 receptor profile gene orders contains in the method detection peripheral blood expanded with PCR
Amount, you can assess the risk that anaphylactoid reaction occurs, it is easy to use, suitable for large-scale crowd examination before patient medication, to promoting
The discovery in advance of crowd easily occurs for anaphylactoid reaction, and the security for improving medication is significant, has broad application prospects.
Further, included provided by the present invention for the gene marker of medicine anaphylactoid reaction sensitive group examination
The characterizing gene sequence of peripheral blood MrgprX2 acceptors shown in SEQ ID NO.2 and SEQ ID NO.3.Present invention also offers
Application of the gene marker in medicine anaphylactoid reaction sensitive group examination product is prepared.Medicine anaphylactoid reaction is sensitive
Mass screening product (i.e. the product of examination anaphylactoid reaction occurrence risk) includes using real time quantitative PCR method examination medicine class mistake
The product of quick reaction sensitive group.And with the product of real time quantitative PCR method examination anaphylactoid reaction occurrence risk comprising special
Property amplification peripheral blood MrgprX2 gene orders primer, its sequence is as shown in SEQ ID NO.2 and SEQ ID NO.3.
Embodiment
Research is found：G-protein on class allergic stimulated thing active cell film first, and then activated protein kinase C and Ca2+It is logical
Road, makes endoplasmic reticulum discharge Ca2+Into intracellular.Along with intracellular Ca2+Concentration increase, mast cell and basophilic granulocyte
Interior granular vesicle moves to endochylema film and discharges Biomedia, so as to trigger anaphylactoid reaction.Class allergic stimulated thing is activated
G-protein be MRGPRS (MAS-Related G Protein-Coupled Receptors), this receptor, can be straight independent of IgE
Mediating mast cell activation is connect, mast cell degranulation is induced.This receptoroid each kind, tissue and organ expression type simultaneously
Differ：Wherein MrgprX2 is expressed in the mast cell of people source；This receptor extracellular region is induction zone, and specificity is not high, when extracellular
After being activated by many kinds of substance such as degranulation polypeptide, Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2, Compound 48/80, beta-alexins, the triggering of intracellular region signal is drawn
Mast cells activation is played, the particulate matters such as histamine are discharged, cell paracrine is stimulated, so as to have influence on peripheral cell.MrgprX2 is
Many small-molecule drugs cause class allergy, the action target spot of false allergy.Therefore by dividing MrgprX2 gene copy numbers
Analysis, so as to deduce expression quantity of the MrgprX2 acceptors in different patients, contributes to evaluation patient that class occurs during medication
Anaphylactoid risk, the security to clinically direction of medication usage is significant.
The present invention is compared grab sample patient and occurred by the gene expression situation of quantitative analysis peripheral blood total serum IgE
The differential expression of peripheral blood MrgprX2 genes in two groups of samples of medicine anaphylactoid reaction patient, then calculated by standard curve
Differential expression occurs in Different Individual peripheral blood for MrgprX2 genes, and (gene expression amount is higher than average level or less than average water
It is flat) gene signal, i.e., the expression signal of MrgprX2 characteristic genes (biomarker) in peripheral blood.Using absolute quantitation
Method, based on peripheral blood MrgprX2 characterizing genes set up patient medication occur anaphylactoid reaction risk profile model.Most
Afterwards, the relative expression quantity of gene marker in person under inspection's peripheral blood is quantitatively detected using fluorescence quantitative RT-RCR, foundation is utilized
Patient medication occurs anaphylactoid reaction risk forecast model and differentiates whether again anaphylactoid reaction and phase can occur after medication for person under inspection
The risk probability answered, instructs clinical application.
Below pair present invention discover that the gene marker for medicine anaphylactoid reaction sensitive group examination it is specific
Process is described in detail.
First, the determination of characterizing gene mark expression quantity baseline in crowd.
Specifically include following steps：
1) 300 are collected using PAXgeneTM Blood RNA Tube heparin tubes and is defined as no disease on inspection and pregnant
The person under inspection's peripheral blood sample for 20-80 the Sui of reaction health of being pregnent, each sample collection 1.5-2mL peripheral bloods.
2) total serum IgE of above-mentioned peripheral blood sample is extracted using Trizol.
(1) 1mL TRIZOL is added in the blood specimen of every 1mL peripheries；
(2) 12000rpm/min is centrifuged 10 minutes, takes 1mL supernatants into 1.5mL EP pipes；
(3) 0.2mL chloroforms are added, are acutely rocked with hand 15 seconds after capping is good, room temperature is placed 2-3 minutes, is then centrifuged for 4
DEG C, 12000rpm/min, 15min.It is divided into three layers after centrifugation, following red is phenol-chloroform phase, an intermediate layer, the above is
Colourless aqueous phase.RNA is existed only in aqueous phase.Aqueous phase accounts for the 60% of total TRIZOL.
(4) upper strata aqueous phase is transferred in another clean EP pipes, adds 0.5mL isopropanols, be stored at room temperature 10min, so
4 DEG C, 12000rpm/min, 10min are centrifuged afterwards.
(5) supernatant is removed, the ethanol washing RNA precipitate of 0.8mL volume fractions 75% is added, centrifuges 4 DEG C, 10000rpm/
Min, 1min, are repeated twice.
(6) supernatant is removed, 0.8mL absolute ethyl alcohols washing RNA precipitate is added, centrifuges 4 DEG C, 10000rpm/min, 1min.
(7) supernatant is removed, is placed in vacuum or air 5-10 minutes, RNA precipitate is dried.With without RNase water or 0.5%SDS
RNA precipitate is resuspended in solution, is blown and beaten repeatedly with pipette tips several times, stands 10 minutes.
(8) concentration is surveyed：5 μ L storing liquids are taken to add in another EP pipe, as blank control, to be surveyed without RNase water
OD260/280 values.1OD=40 μ g RNA.It is very high that OD260/280 values are considered as purity in 1.8-2.0.It is general dense in 1000ng/mL
It is calibrated.Concentration too it is high to dilute after determine again.
(9) RNA is identified：Denaturing formaldehyde agarose electrophoresis, determines extracting RNA integrality and DNA pollution situation.
3) purity of RNA samples is detected with Bio-Tek Epoch microplate spectrophotometers.All RNA samples must be accorded with
Close following Quality Control condition：RNA yields are more than 2 micrograms, and the ratio at 28S/18S peaks is more than 1, RIN values and is more than 7,260nm/280nm's
Absorbance ratio is big by 1.8.
4) the PrimeScriptTM RT reagent Kit with gDNA Eraser of Takara companies are utilized
(Perfect Real Time) kit removes the genome in total serum IgE, and carries out reverse transcription.
5) real-time fluorescence quantitative PCR detection cycles samples number is carried out using standard plasmid, and draws standard curve, to amplification
Rear sample carries out copy number detection, and tries to achieve using standard curve the copy number of sample.
Wherein quantitative fluorescent PCR condition is as shown in table 1.
The quantitative fluorescent PCR condition of table 1
Wherein, the sequence of Forward primers is：5'-caggacattgctgaggtgga-3'；
The sequence of Reverse primers is：5'-agttcagcaaatcagacagacagg-3'.
6) statistical method is utilized, statistical analysis is carried out to 300 samples, 95% confidential interval and normal state point is calculated
Cloth situation.
2nd, the determination of characterizing gene mark expression quantity in allergic human population.
Specifically include following steps：
1) the 20-30 peripheral blood samples for clinically determining to occur anaphylactoid reaction patient, each sample collection are collected
1.5-2mL peripheral blood.
2) according to the step in above-mentioned the determination of characterizing gene mark expression quantity baseline in crowd " one, ", sample is extracted
Product total serum IgE and the copy number for detecting MrgprX2 genes.
3) 95% confidential interval and normal distribution feelings of allergic human population's gene copy number are calculated using statistical method
Condition.
4) copy number of MrgprX2 genes in statistical method, two class crowds of contrast is utilized, the difference of copy number is found out
Property.
3rd, result.
1st, to general population and allergic disease, everybody analyzes collection blood sample number of mining massively, as a result as shown in table 2.
The general population of table 2 and the analysis of allergic disease people crowd's blood sample number
2nd, sequence table
The characterizing gene sequence of MrgprX2 acceptors is as shown in SEQ.ID.NO.1.
Primer sequence is as follows：
SEQ.ID.NO.2：Forward primers：5'-caggacattgctgaggtgga-3'；
SEQ.ID.NO.3：Reverse primers：5'-agttcagcaaatcagacagacagg-3'.
3rd, basophilic granulocyte MrgprX2 expression of receptor is detected
Basophilic granulocyte total serum IgE in whole blood is extracted, MrgprX2 gene expression amounts, knot are detected using RT-PCR method
As shown in Table 3, it is about (1.1 ± 0.23) × 10 to measure MrgprX2 gene copy numbers to fruit4, illustrate that MrgprX2 acceptors are thermophilic in whole blood
Expressed on alkaline granulocyte.
The basophilic granulocyte gene copy number of table 3
In addition, having carried out Western Blot detections and whole blood immunity fluorimetric analysis.As a result it is as shown in table 4, from
The basophilic granulocyte that Western Blot results can be seen that in whole blood in table 4 is able to detect that MrgprX2 receptor proteins
Expression.
MrgprX2 receptor proteins content Western Blot are detected in the whole blood of table 4
In addition, immunofluorescence results are shown, basophilic granulocyte in whole blood can be positioned using specific antibody CD63,
Further it is marked using the antibody of MrgprX2 receptor-specifics, specify that MrgprX2 acceptors are true in basophilic granulocyte
Real storage.
4th, basophilic granulocyte functional selection
The functional experiment of calcium imaging and histamine release is carried out using basophilic granulocyte KU812 cell lines in blood.Knot
Fruit shows that the activator of MrgprX2 receptor-specifics, C48/80, Ciprofloxacin, morphine and atracurium can activate two kinds
The calcium mobilization of cell, causes the rise of cell intracellular free calcium level.Meanwhile, histamine release shows, 4 kinds of activators are responsible for
KU812 cells release histamines, and burst size and dosage are into positive correlation.
Basophilic granulocyte in above the results show, whole blood functionally has certain related to mast cell
Property, two kinds of cells have certain homology.Therefore it can be detected loose thin to reflect to basophilic granulocyte in whole blood
The expression of receptor situation of born of the same parents.Simply, the difference of patient's MrgprX2 expression of receptor is easily and efficiently evaluated, is the wind of clinical application
It is dangerous to be evaluated.
The gene marker for patient's generation anaphylactoid reaction Risk Screening of the offer of the present invention, can be used in class mistake
The quick occurrence risk examination that reacts, and obtained in terms of the product of examination early stage medicine anaphylactoid reaction sensitive group is prepared
Using, there is sensitiveness height, high specificity, and due to being to be used as detection sample using the peripheral blood clinically most easily gathered
This, it is easy to detect, it is adaptable to the large-scale crowd examination and early diagnosis of medication patient and medicine anaphylactoid reaction sensitive group,
Have a extensive future.
It is described above, only it is presently preferred embodiments of the present invention, not the present invention is imposed any restrictions, it is every according to the present invention
Any simple modification, change and equivalent structure transformation that technical spirit is made to above example, still fall within skill of the present invention
In the protection domain of art scheme.
Nucleotides sequence list
<110>Xi'an Communications University
<120>A kind of gene marker and its application for medicine anaphylactoid reaction sensitive group examination
<160> 3
<210> 1
<211> 6227
<212> DNA
<213> MrgprX2
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gtttgccagt cccaggaaag cacttctcaa ctcaccaact ccagtagaaa gaagggtgtt 60
aaggtaagag tttgttcaag aaccatcttc tttcaaaggc agttttggtt tttaccttag 120
tccacatcct gatgaattca ttaggaaaag aagacaggaa gctcctttga ggaagctttg 180
gatgctggca gcttagatgc tgggagatcc gaatggggtc aagcaggttc gtcccacttc 240
tcttctatct accaatattc tccccttctc ttcgaccatt caaatctgaa actatggaga 300
gaaagatttt gctgatttta atttaggggc aggctagaat gaagtcatta cttccctaag 360
ctaaattgtt atttaaaacc tcaggttgca ctgattttct tgttattcaa aaggtttttc 420
cttcatatct gtcattgtcc cagcagaaaa cacacatcac acactctaaa ggggtagttt 480
agaggagttt agtgaagaga ctgtttacaa cggcgtgaac agggattaag gaatgatgaa 540
gccctcaaag gcaagtgata gtggagggct gttaccgttg ctaggtctga agtgataagg 600
agaaagagaa gttctgagaa ctcagaatgt gctgtagcgg taactgtaac tgtaactata 660
gctgtggcta caggaaaggg ccaccaggca gggctatgtc cttaggtaga aaaacactgc 720
cactgccaac tcacagccct tcagggcgca gggagagagc caggaaattt ttaaaaaatc 780
atcccccaat ctactgtcaa tgtgtccctt tggctgaaaa aaaaagtcac cctccaatct 840
cctgtcaatg tgtacccttt ggagcctgag tgaaagacag cccattgacg aggcacagac 900
atgtctcctc ccaggatgca aaggcaggta gagaaggatg gacatgagat cctaatagca 960
aataggaaag tccatttctt cccatagcct cttctcagtt tgtctttcct gagactttct 1020
ctattaatgt gattgaatca atttctcatt ctatcacctc cctttttttt aattgtttga 1080
ccttcccatc tagtgtcact tctttggact agtctctcac tatcatcata aatgccttga 1140
gaatggaatg tggttgggaa aaaaagggat tgggagtaca taggtactcc cagctataag 1200
tacacaggta tgtgtaatgt aagtttatga ttttggcttc tctaaaaaga aacctattat 1260
agtgataatc aaaaagaatg tttactagac ttgggctgat aactctaaga catcaaacat 1320
caaggctcct tccaatttct atttccccat ccctaaagcg gtacccttac cctcatggtc 1380
ctacatggct cccaaacaca tcagcattcc agccagagag aagagaaaaa aagaaagagt 1440
acaggagtag ctggttatat cactttctct cacctcccct tgaccatcac atatggttac 1500
atggtgccat tagcataaag ggaagctgag aaatgggcct ttttctgatc taagactcag 1560
aattattatg tgtgagaaaa ggagaatggc ttttggggca acaagcagtc tctatagccc 1620
gattctcttt tagcatctgt gctcacctgg agatttttct caacactgcc tcacaaatag 1680
tgaatgatgt agacatggaa cagaaataaa cctaggacag ggccccagga gactggagct 1740
ggtattggac ctgcttttca ccatgtgatt caggaaactc ttttgtacac gttaagcctg 1800
tctttctttg ttaaatgagc gtattacaat agatggcctc caagggatat gtgttggagg 1860
ctggggatat aatagtgaac aaccacaaca aaaatgttga gcttaatgag ggaagtaaaa 1920
gaaaaaacag acatacactt aaaaatacag gtgcagggcc gggcacggtg gctcacgcct 1980
gtaatcctgg cactttggga ggccgaggcg ggcagatcac aaggtcagaa gttcgagacc 2040
atcctggcta acacagtgaa accccgtctc tactgaaaaa aaaaatacaa aaaattagcc 2100
agtcgtggtg gtgggcacct gtagtcccag ctactcggga ggctgaggca ggagaatggc 2160
gtgagcccag gaggcggagc ttgcagtgag ccgagatcga gccactgcac tccagcctgg 2220
gtgacagagc gagactccat ctcaaaaaaa aaaaaaaata caggtgcaaa ctgtgttcag 2280
tgccacgaat aaaaaggcca acctgccctg agggaggaca acagaagtgg ttttcttcca 2340
tatcctcaca aggccaacca gggctacagg aactaagaca gcatgtgatg gatgcattca 2400
gtccttgatg ccaattctgc acccccatgg aggacacatt cactttcccc ttggaggcct 2460
ccaacccatg agagttttct gcagtgtgga gtgattgacg gctgtttttc tgaagcaaga 2520
aagagctctt gcattcagtt gcttattctc agttggcttc tattcatatc tcttctcctt 2580
atttgattaa tctctcttcc ttactgtttc ttaaagttgc ttaatctagg cttaaaaaac 2640
tgcaatttcg acattccatt ttggatttga tatgagcatt acttgttgct tttctccttt 2700
cttcatgtgt agttggtatt gaacccttgc aatggggctg caaggctggt taagaacaag 2760
ccctctgaca cagacctgtg ggttcaaatg caggctctac cacttgtttg tgagacctga 2820
agcatattac tcaatacatt tgtgctttaa tctccttctc tgtaaaatgg gactaataat 2880
agtactgacc tcaccaggct ctgaagaaga tttaaagagg ctatacctga agagcattca 2940
gccagctgct attattcacc ccattatgag taagggctcc ttcccaagtg taatttgcat 3000
aagcatctgc cttttcccct caaagccctg cacagtgtag acatggttct tttttctaat 3060
ttcccactcc tttccccaaa taatcctgtt cacaaatgct ttaatttgca aagaaagctc 3120
aaaatcatct gggggaagta atgggtccac cttagatgta ttcataccac aacaccctct 3180
agttacctct agacactaga cactctcaat tgctgcctct ctctcctttt cccaaattgc 3240
ccccttttcc tccaagatgc agagaaattc agataaaccc tgaatcctct ccataaactg 3300
gaagtttaaa ctcttttggg tgctgagcaa aagccatata ctagcccttc ctggattgaa 3360
gatatgacag tggccattat tcactatctg tgtaggcaga agcaatccct tcaaatacac 3420
cagagctttc ccagaaggaa acatgaagaa tgatgttttc ttcatttctt tacatgtgtg 3480
ttttttagtg actaatttca ttttaaaatg ttggtgttcc acttagccca aattttcaac 3540
tactcttggc tccccatgca tgcatgcatg cattcattca ttcatttata tgtggtacaa 3600
acttcttgac cctccactat gccctcacat tgagctaggc actgaggaca cagaaatgga 3660
taagagaaaa actctttctg ttttcataaa gctgacccac tgagcaattt ctagtctgac 3720
aggcctggct gtcctcagac ctgtcctagc aagaagtcac ctctccgtta tgaccctaaa 3780
tcacaatctc cattctgact gttgatggac tcctctcagt cattaagggt ggccatgggt 3840
catggcctgg gtcatttctt ctggcccctg agaggaaatc tgtatgccag gataaaaaga 3900
atcctgaact cagagtactg tctcttgctg ccccacatat tctgcctaga cttttctctt 3960
cctgcctcca ggagacatga cacagtgaca gtgagtgggg gtgtttgggc cttagaatat 4020
tcccatacca gcagaggatc taccttgtaa tagaaagagc ccaggattta gagtcagcaa 4080
gaaatgagtt tgaatccagg tgctaggact ccctggccct taataaatga cttaatctct 4140
tcaagcctct gatttcctct cctgtaaaac aggggcggta attaccacat aacaggctgg 4200
tcatgaaaat cagtgaacat gcagcaggtg ctcaagtctt gtttttgttt ccaggggcac 4260
cagtggaggt tttctgagca tggatccaac caccccggcc tggggaacag aaagtacaac 4320
agtgaatgga aatgaccaag cccttcttct gctttgtggc aaggagaccc tgatcccggt 4380
cttcctgatc cttttcattg ccctggtcgg gctggtagga aacgggtttg tgctctggct 4440
cctgggcttc cgcatgcgca ggaacgcctt ctctgtctac gtcctcagcc tggccggggc 4500
cgacttcctc ttcctctgct tccagattat aaattgcctg gtgtacctca gtaacttctt 4560
ctgttccatc tccatcaatt tccctagctt cttcaccact gtgatgacct gtgcctacct 4620
tgcaggcctg agcatgctga gcaccgtcag caccgagcgc tgcctgtccg tcctgtggcc 4680
catctggtat cgctgccgcc gccccagaca cctgtcagcg gtcgtgtgtg tcctgctctg 4740
ggccctgtcc ctactgctga gcatcttgga agggaagttc tgtggcttct tatttagtga 4800
tggtgactct ggttggtgtc agacatttga tttcatcact gcagcgtggc tgattttttt 4860
attcatggtt ctctgtgggt ccagtctggc cctgctggtc aggatcctct gtggctccag 4920
gggtctgcca ctgaccaggc tgtacctgac catcctgctc acagtgctgg tgttcctcct 4980
ctgcggcctg ccctttggca ttcagtggtt cctaatatta tggatctgga aggattctga 5040
tgtcttattt tgtcatattc atccagtttc agttgtcctg tcatctctta acagcagtgc 5100
caaccccatc atttacttct tcgtgggctc ttttaggaag cagtggcggc tgcagcagcc 5160
gatcctcaag ctggctctcc agagggctct gcaggacatt gctgaggtgg atcacagtga 5220
aggatgcttc cgtcagggca ccccggagat gtcgagaagc agtctggtgt agagatggac 5280
agcctctact tccatcagat atatgtggct ttgagaggca actttgcccc tgtctgtctg 5340
atttgctgaa ctttctcagt cctgatttta aaacagttaa gagagtcctt gtgaggatta 5400
agtgagacag tgcctatgaa acaaacacta agtgcagtgt ctctggaact gccttactca 5460
caggcttcca ccacagccct atgagagctt tgccaactct gcggtccatg actgttccca 5520
cttttaatga atcctacctt tcgcagaagg ctgaaagcag ggcagaaaag atctacattt 5580
ctttggacac tgcacttgat agggactcaa agaatgttat atttttaatt aatttctttt 5640
tctcttccgt acaatttctg tctcaacaaa attagaagaa ttaaatttaa aactagctcc 5700
aaaagagcag tcgtctttca ttttggcaga ccttagaata tccccctagc ttaataaatc 5760
tttgttgaat ggcttaatga atgaataaac tggttaatgt ttaagttaaa cctctgaaaa 5820
gtctccattt accagatttg agtcactaaa tttattgctt tcactacttt tgaattttgc 5880
aaacatgaaa ttaagtttta taattagata aatcaatgtc aacacatatt taaagtttga 5940
ggtacactgt cttcctgtgg tttcctttca catgccatcc cttaaaatcc cagctacacg 6000
ccttcccatt ccttcccctt tgcctttgtt ctaatcttcc ctctctgggg gctctctaat 6060
tcgtcctgga agtttccagt ggtcttatag actccatgtt cttggaggac aggctgtatg 6120
tcagatttac cttttattcc gaagaactcg gagcatttat tttgttaatt aaattgcaca 6180
tatttttaaa agttacgtgt tccacagaat aaaatactaa ttgtaaa 6227
<210> 2
<211> 20
<212> DNA
<213>Forward primers
<400> 2
caggacattg ctgaggtgga 20
<210> 3
<211> 24
<212> DNA
<213>Reverse primers
<400> 3
agttcagcaa atcagacaga cagg 24
Claims (6)
1. a kind of gene marker for medicine anaphylactoid reaction sensitive group examination, it is characterised in that：The genetic marker
Thing is the characterizing gene sequence of MrgprX2 acceptors.
2. the gene marker according to claim 1 for medicine anaphylactoid reaction sensitive group examination, its feature exists
In：The characterizing gene sequence of the MrgprX2 acceptors is as shown in SEQ ID NO.1.
3. the gene marker for medicine anaphylactoid reaction sensitive group examination described in claim 1 or 2 is preparing medicine
Application in anaphylactoid reaction sensitive group examination product.
4. application as claimed in claim 3, it is characterised in that：The medicine anaphylactoid reaction sensitive group examination product includes
With the product of real time quantitative PCR method examination medicine anaphylactoid reaction sensitive group.
5. application as claimed in claim 4, it is characterised in that：The use real time quantitative PCR method examination medicine class allergy is anti-
Answer the primer of characterizing gene sequence of the product comprising specific amplification MrgprX2 acceptors of sensitive group.
6. application as claimed in claim 5, it is characterised in that：The primer includes Forward primers and Reverse primers,
The wherein sequence of Forward primers is as shown in SEQ ID NO.2, and the sequence of Reverse primers is as shown in SEQ ID NO.3.
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| CN201710509594.2A CN107142325A (en) | 2017-06-28 | 2017-06-28 | A kind of gene marker and its application for medicine anaphylactoid reaction sensitive group examination |
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|---|---|---|---|
| CN201710509594.2A CN107142325A (en) | 2017-06-28 | 2017-06-28 | A kind of gene marker and its application for medicine anaphylactoid reaction sensitive group examination |
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| CN107142325A true CN107142325A (en) | 2017-09-08 |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107703312A (en) * | 2017-11-15 | 2018-02-16 | 西安交通大学 | The protein marker and method of examination medicine anaphylactoid reaction sensitive group |
| CN110204606A (en) * | 2019-05-31 | 2019-09-06 | 西安交通大学 | A kind of polypeptide being used to prepare rabbit polyclonal antibody and its application |
| CN110204607A (en) * | 2019-05-31 | 2019-09-06 | 西安交通大学 | A kind of dominant antigen epitope polypeptide of anti-Mrgprx2 antibody and its application |
| CN110229228A (en) * | 2019-05-31 | 2019-09-13 | 西安交通大学 | The polypeptide with immunogenicity of a kind of pair of anaphylactoid reaction specific receptor MRGPRX2 albumen and its application |
| CN110229227A (en) * | 2019-05-31 | 2019-09-13 | 西安交通大学 | It is a kind of be used to prepare ELISA murine monoclonal coated antibody and rabbit polyclonal detection antibody polypeptide and its application |
| CN110240643A (en) * | 2019-05-31 | 2019-09-17 | 西安交通大学 | A kind of double antibodies sandwich kit being used to prepare anaphylactoid reaction matches polypeptide and its application of antibody |
| WO2021201486A1 (en) * | 2020-03-31 | 2021-10-07 | 재단법인 아산사회복지재단 | Method for identifying patient with immediate hypersensitivity reactions by using mrgprx2 |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107703312A (en) * | 2017-11-15 | 2018-02-16 | 西安交通大学 | The protein marker and method of examination medicine anaphylactoid reaction sensitive group |
| CN110204606A (en) * | 2019-05-31 | 2019-09-06 | 西安交通大学 | A kind of polypeptide being used to prepare rabbit polyclonal antibody and its application |
| CN110204607A (en) * | 2019-05-31 | 2019-09-06 | 西安交通大学 | A kind of dominant antigen epitope polypeptide of anti-Mrgprx2 antibody and its application |
| CN110229228A (en) * | 2019-05-31 | 2019-09-13 | 西安交通大学 | The polypeptide with immunogenicity of a kind of pair of anaphylactoid reaction specific receptor MRGPRX2 albumen and its application |
| CN110229227A (en) * | 2019-05-31 | 2019-09-13 | 西安交通大学 | It is a kind of be used to prepare ELISA murine monoclonal coated antibody and rabbit polyclonal detection antibody polypeptide and its application |
| CN110240643A (en) * | 2019-05-31 | 2019-09-17 | 西安交通大学 | A kind of double antibodies sandwich kit being used to prepare anaphylactoid reaction matches polypeptide and its application of antibody |
| CN110229228B (en) * | 2019-05-31 | 2021-05-28 | 西安交通大学 | Polypeptide with immunogenicity to anaphylactoid reaction specific receptor MRGPRX2 protein and application thereof |
| CN110240643B (en) * | 2019-05-31 | 2021-05-28 | 西安交通大学 | Polypeptide for preparing anaphylactoid reaction double-antibody sandwich kit paired antibody and application thereof |
| CN110229227B (en) * | 2019-05-31 | 2021-05-28 | 西安交通大学 | Polypeptide for preparing ELISA mouse monoclonal coating antibody and rabbit polyclonal detection antibody and application thereof |
| WO2021201486A1 (en) * | 2020-03-31 | 2021-10-07 | 재단법인 아산사회복지재단 | Method for identifying patient with immediate hypersensitivity reactions by using mrgprx2 |
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Application publication date: 20170908 |