CN110277138A - A kind of steroid femur head necrosis tumor susceptibility gene detection model - Google Patents
A kind of steroid femur head necrosis tumor susceptibility gene detection model Download PDFInfo
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Abstract
The invention discloses a kind of steroid femur head necrosis tumor susceptibility gene detection models, its are as follows: F=human genome site rs2305027 scoring+site rs34433978 scoring+site rs3833221 scoring+site rs143471015 scoring, wherein F is steroid femur head necrosis risk factor；It is steroid femur head necrosis high risk as F >=5；It is steroid femur head necrosis low-risk as F≤4, risk is positively correlated with score.Doctor can be used for using the detection model and assess patient if appropriate for using glucocorticoid medicine to treat, estimate its risk that caput femoris necrosis occurs, to instruct the clinical use of glucocorticoid medicine, implement precisely medical treatment.
Description
Technical field
The invention belongs to genetic test fields, and in particular to a kind of steroid femur head necrosis tumor susceptibility gene detection model,
It further relates to a kind of for detecting the kit of steroid femur head necrosis tumor susceptibility gene.
Background technique
Caput femoris necrosis (Osteonecrosis of the femoral head, ONFH), also known as ischemic femoral head is bad
Extremely (Avascular necrosis, AVN), refers to blood circulation obstacle in femoral head caused by a variety of causes, leads to osteocyte
Anoxic, shrinkage and death (ZALAVRAS C G, LIEBERMAN J R.Osteonecrosis of the femoral head:
Evaluation and treatment.J Am Acad Orthop Surg, 2014,22 (7): 455-464.), cause hip to close
Save pain and moving obstacle.In the numerous reasons for leading to ONFH, the caput femoris necrosis (SiONFH) of steroids induction is considered
It is one of mostly important ONFH type in addition to traumatic ONFH (MONT M A, CHERIAN J J, SIERRA R J, et
Al.Nontraumatic Osteonecrosis of the Femoral Head:Where Do We Stand Today? A
Ten-Year Update.J Bone Joint Surg Am,2015,97(19):1604-1627.).Pathology about SiONFH
There are a variety of hypothesis for mechanism, including bone microcirculation blood supply disorder, blood coagulation disorders, Derived from Mesenchymal Stem Cells are abnormal, lipid metaboli is different
Often etc., but still different patients can not be explained to the individual difference of SiONFH neurological susceptibility completely.Ono etc. (ONO K, TOHJIMA T,
KOMAZAWA T.Risk factors of avascular necrosis of the femoral head in patients
with systemic lupus erythematosus under high-dose corticosteroid therapy.Clin
Orthop Relat Res, 1992 (277): 89-97.) 62 systemic loupus erythematosus (SLE) patients of follow-up, though receive high
Dose glucocorticoid treatment, but do not find hormone daily dosage, intergal dose and significant phase occurs using duration and SiONFH
It closes, even there is the patient but to necrose using low dosage hormone, this part population can not for the neurological susceptibility of SiONFH
It is explained from hormone dosage, thus it is speculated that there may be genetic correlation factors.Therefore, genetic predisposition is that the another of SiONFH is ground
Study carefully hot spot.Even if using similar hormone scheme in same primary disease, however it remains the individual difference of SiONFH disease incidence
It is different.(LAUSTEN G S, JENSEN J S, the OLGAARD K.Necrosis of the femoral head after such as Yang
Renal transplantation.Acta Orthop Scand, 1988,59 (6): 650-654.) by SLE patient
Research finds that C3435T or the G2677T variation of MDR1 gene may participate in SLE by the expression and function for influencing P-gp albumen and suffer from
The development of SiONFH in person；However, the verifying in different queue has been found that its consistency is not high.It is not yet clearly any at present
The genetic susceptibility loci of one SiONFH.Single nucleotide polymorphism (Single Nucleotide Polymorphisms,
It SNP) is the most common mutant dna sequence.Its definition is the variation of single nucleotide acid in the genome, and this variation can be by single
The conversion (transition) or transversion (transversion) of base are caused, can also be caused by the insertion of base or missing.
Previously research fails to screen the hereditary difference using SNP as representative with accomplishing high-throughput wide spectrum, therefore obtained susceptibility loci
Very limited, the verifying rate in different crowd is lower, can not provide valuable reference to clinical diagnosis.As science of heredity is ground
Study carefully the development of technology, new-generation sequencing technology (NGS) cost gradually decreases, and precision is gradually increased, science of heredity detection depth and
Range continues to increase, it will be found that a greater amount of genetic susceptibility locis provides more to disclose the pathogenesis of SiONFH
The SiONFH diagnosis and treatment scheme of fining.
Summary of the invention
So far, there is no literature reported on carry out detection and hormonal femoral head using high throughput sequencing technologies perspectively
Thus the relevant gene of necrosis or gene set simultaneously optimize or select therapeutic scheme.In order to fill up the blank of the clinical technology, reach
Precisely medical treatment, the administration accuracy for ensureing glucocorticoid user, the purpose of raising success rate, inventor combines clinical
Practice and gene sequencing technology find some steroid femur head necrosis tumor susceptibility genes, and based on the polymorphic of these tumor susceptibility genes
Property establishes a kind of steroid femur head necrosis tumor susceptibility gene detection model, which is a kind of mathematical model, can be used for assessing
A certain patient reduces the risk that caput femoris necrosis occurs for patient if appropriate for using glucocorticoid to be treated.It is specific and
Speech, the present invention includes following technical solution.
A kind of steroid femur head necrosis (SiONFH) tumor susceptibility gene detection model, are as follows: F=human genome site
Rs2305027 scoring+site rs34433978 scoring+site rs3833221 scoring+site rs143471015 scores, wherein F
For steroid femur head necrosis (SiONFH) risk factor, it is positively correlated pass with steroid femur head necrosis (SiONFH) risk
System.
In scoring (score) mode in above-mentioned each site, the standards of grading of site rs2305027 are: rs2305027CC
That is two allele are C base in the site, are denoted as 0 point；One of rs2305027CT i.e. two allele is in the site
C base mutation be T base, be denoted as 1 point；C base of rs2305027TT i.e. two allele in the site sports T
Base is denoted as 2 points；
The standards of grading of site rs34433978 are: rs34433978GG i.e. two allele is G alkali in the site
Base is denoted as 0 point；G base of one of rs34433978G- i.e. two allele in the site is lost, and is denoted as 1 point；
G base of rs34433978-- i.e. two allele in the site is lost, and is denoted as 2 points；
The standards of grading of site rs3833221 are: the rs3833221-- i.e. two allele site is inserted into without base,
It is denoted as 0 point；One of the rs3833221C- i.e. two allele position inserts a C base, is denoted as 1 point；
The rs3833221CC i.e. two allele site inserts a C base respectively, is denoted as 2 points；
The standards of grading of site rs143471015 are: rs143471015GG i.e. two allele is G alkali in the site
Base is denoted as 2 points；The G base deletion of one of rs143471015G- i.e. two allele in the position, is denoted as 1 point,
The G base of rs143471015-- i.e. two allele in the position lacks, and is denoted as 0 point.
It is steroid femur head necrosis high risk as F >=5, indication clinically should cautiously make the patient in the model
With glucocorticoid medicine, including reduces dosage, uses the measures such as time；It is that steroid femur head necrosis is low as F≤4
Risk, indication are smaller a possibility that clinically the patient is necrosed using glucocorticoid medicine.
Since the model is a kind of mathematical model, can be entered gene detection equipment message processing module,
Or it is entered the message processing module of blood testing equipment.
In one embodiment, above-mentioned steroid femur head necrosis tumor susceptibility gene detection model can be entered intelligent doctor
It treatment system or is entered in the computer of doctor.
Another aspect of the present invention is to provide a kind of for detecting the reagent of steroid femur head necrosis tumor susceptibility gene
Box comprising be respectively used to the primer sequence of detection gene rs2305027, rs34433978, rs3833221 and rs143471015
It arranges (or primer pair sequence), carries out amplification and subsequent sequencing detection for use in above-mentioned site.
In a preferred embodiment, the above-mentioned primer pair for detecting gene rs2305027 is forward primer SEQ
ID NO:1 and reverse primer SEQ ID NO:2；Primer pair for detecting gene rs34433978 is forward primer SEQ ID
NO:3 and reverse primer SEQ ID NO:4；Primer pair for detecting gene rs3833221 is forward primer SEQ ID NO:5
With reverse primer SEQ ID NO:6；For detect gene rs143471015 primer pair be forward primer SEQ ID NO:7 and
Reverse primer SEQ ID NO:8.
For detecting the amplification of gene rs34433978, rs2305027, rs143471015 and rs3833221 by PCR
Primer sequence is not limited to above-mentioned sequence SEQ ID NO:1-8, it is clear that including other any suitable primers.
When carrying out PCR amplification to said gene using mentioned reagent box, PCR reaction system can be with are as follows: forward primer, 10 μ
M, 1 μ L；Reverse primer, 10 μM, 1 μ L；2xTaq Master Mix, 10 μ L；DNA sample, 1 μ g；Add ddH2O to 20 μ L, and
PCR reaction condition are as follows: 95 DEG C of 5min；(95 DEG C of 30sec, Tm DEG C of 30sec, 72 DEG C of 1min) × 35 are recycled；72℃7min；4 DEG C of guarantors
Temperature.
It, can be with when implementing genetic test to the target group for needing to carry out SiONFH risk assessment using mentioned reagent box
Its full-length genome sample (including saliva, blood, body fluid etc.) is acquired using various existing methods.For SNP site
Rs34433978, rs2305027, rs143471015 and rs3833221 design amplimer sequence, carry out PCR reaction, and right
Amplified production carries out the sequencing of Sanger method (or other reliable genetic methods), with clearly each susceptibility loci multistate character
State.
Susceptibility loci polymorphism status is inputted into above-mentioned steroid femur head necrosis tumor susceptibility gene as variable and detects mould
It is gone after type, the clinical risk coefficient that SiONFH occurs for patient can be calculated.
Above-mentioned steroid femur head necrosis tumor susceptibility gene detection model is a kind of integrated diagnosis model comprising 4 variables,
Or risk evaluation model can be to calculate due to being a kind of mathematical model for glucocorticoid administration risk profile
Machine software (computer program) form is entered intelligent medical system or is entered in the computer of doctor, helps doctor's judgement
The clinical risk of SiONFH occurs for Patients with Aseptic Necrosis of Femoral, assesses it if appropriate for using glucocorticoid to be treated, instructs
The clinical use of glucocorticoid medicine implements precisely medical treatment.
Steroid femur head necrosis tumor susceptibility gene detection model provided by the invention is in Zhongshan Hospital Attached to Fudan Univ
It is verified in clinical practice, retrospective study (including 23 necrosis and 15 check samples) knot based on 38 samples
Fruit draws area (AUG) under ROC curve calculated curve, and the AUC of this model is 0.835 as the result is shown, sensitivity (or it is sensitive
Degree) it is 87%, specificity (or specificity) is 80%, has Clinical significance of MG.
Detailed description of the invention
Fig. 1 shows the ROC curve that the prediction model constructed to the present invention is verified, and wherein AUC=0.835, sensitive
Degree 87%, specificity 80%.
Specific embodiment
It is susceptible that the present invention from science of heredity angle has investigated the caput femoris necrosis (SiONFH) that different patients induce steroids
The individual difference of property, by in human genome largely may gene relevant to SiONFH screened, it was found that
Wherein at least 4 genomic locus rs2305027 (being related to functional gene COLEC12), rs34433978 (are related to functional gene
CYP2B6, CYP2A13), rs3833221 (being related to functional gene CYP2F1), rs143471015 (be related to functional gene
SLFN12L) highly relevant, the mutation of their Individual base forms gene pleiomorphism, this polymorphism (including base mutation
Quantity, mutation type) influence for SiONFH is different.According to this polymorphism, using this 4 gene locis as variable
Amount constructs a kind of mathematical model.
Mathematical model accordingly, it can be estimated that it is relevant that glucocorticoid occurs using the patient that glucocorticoid is treated
The clinical risk of caput femoris necrosis then instructs the clinical use of glucocorticoid medicine.That is, the invention proposes for facing
Accurate evaluation particular patient is difficult to if appropriate for a kind of solution for using glucocorticoid treat this problem in bed practice
Method.
It is obvious to one skilled in the art that steroid femur head necrosis tumor susceptibility gene detection model may be it
His mathematical model form.
Herein, term " genomic locus ", " site " in site in human genome relevant to SiONFH are addressed
Or " SNP site " indicates identical meaning, may be used interchangeably, referring in particular to can be with steroid femur head necrosis (SiONFH)
Such as steroids induction caput femoris necrosis it is relevant can mutational site can variant sites.
Mathematical model of the invention can be entered gene detection equipment such as by programming, by mathematical software packet form
The message processing module of gene sequencer or the message processing module for being entered blood testing equipment；Cloud can also be entered
Server forms sharable intelligent medical system or trouble can be generated in the computer for being entered doctor, the operation of these equipment
The instruction of person's generation steroid femur head necrosis risk size.
In mathematical model of the invention, the mutation type in order to show the i.e. allele of gene polynorphisms is different, adopts
With diacritic or base after genomic locus rs2305027, rs34433978, rs3833221, rs143471015
The representation of (being selected from A, C, G, T) as suffix.The number form is international to SNP (Single nuclear polymorphism)
The mode being labeled is numbered by the reference SNP that NCBI (US National Biotechnology Information center) summarizes confirmation, passes through the volume
Number can be convenient ncbi database (http://www.ncbi.nlm.nih.gov/) in retrieve the related letter of the SNP
Breath.Additional symbol or base after rs number (A, C, G, T, -), it is phonetic to respectively represent adenine, cytimidine, guanine, thymus gland
Pyridine, without five kinds of meanings of base.The combination of two symbols or base respectively represents two equipotential bases in diplont body cell
Because of respective base state.Representing two allele corresponding positions such as " TT " is T base；" CT " represents two allele
Middle corresponding position one is C base, and one is T base；It is G base that " G- ", which represents position one in two allele, separately
One is vacancy, specifically represents insertion mutation or deletion mutation, and retrieval in the database need to be numbered according to rs and is assert.
Above-mentioned site rs2305027 refers to that the 334742nd C base mutation of No. 18 chromosomes is T base.Site position
In Colec12 gene extron sub-district, the protein function that will lead to gene coding is abnormal.Due to the presence of allele,
The site rs2305027 is there are CC, tri- kinds of states of CT, TT, i.e. CC refers to that two allele in the site are C base, and CT refers to two
C base mutation of one of a allele in the site is T base, and TT refers to that C base of two allele in the site is prominent
Become T base.In scoring (score) mode of model of the present invention, rs2305027CC is denoted as 0 point, and rs2305027CT is denoted as 1
Point, rs2305027TT is denoted as 2 points.
Rs34433978 refers to the 41567532nd G base deletion mutation of No. 19 chromosomes.The site is located at CYP2B6,
CYP2A13 intergenic region leads to the section frameshift mutation, seriously affects gene function.Due to the presence of allele,
The site rs34433978 there are GG, G-, -- three kinds of states, i.e. GG refer to that two allele in the site are G base, and G- refers to two
G base of one of a allele in the site is lost, -- refer to that G base of two allele in the site is lost.At this
In scoring (score) mode of invention model, 34433978 GG of rs is denoted as 0 point, and 34433978 G- of rs is denoted as 1 point, rs
34433978-- is denoted as 2 points.
Rs3833221 refers to C base insertion mutations at No. 19 the 41622107th bit bases of chromosome.The site is located at
CYP2F1 gene extron sub-district leads to the section frameshift mutation, seriously affects the protein function of gene coding.Due to equipotential base
The presence of cause, 3833221 site rs exist --, tri- kinds of states of C-, CC, i.e., -- refer to two allele in the site without base
Insertion, C- refer to that one of two allele insert a C base in the position, and CC refers to that two allele are distinguished in the site
Insert a C base.In scoring (score) mode of model of the present invention, rs 3833221-- is denoted as 0 point, rs 3833221
C- is denoted as 1 point, and 3833221 CC of rs is denoted as 2 points.
Rs143471015 refers to that deletion mutation occurs for G base at No. 17 the 33806842nd bit bases of chromosome,
The site rs3833221 there are GG, G-, -- three kinds of states, i.e. GG refer to that two allele in the site are G base, and G- refers to two
The G base deletion of one of a allele in the position, -- refer to that the G base of two allele in the position lacks.At this
In scoring (score) mode of invention model, rs143471015-- is denoted as 0 point, and rs143471015 G- is denoted as 1 point,
Rs143471015 GG is denoted as 2 points.
It is easy that steroid femur head necrosis of the invention is demonstrated in the clinical practice that Zhongshan Hospital Attached to Fudan Univ carries out
Feel the reliability of genetic test model.
In order to make the present invention more obvious and understandable, it is hereby described in detail below with embodiment cooperation attached drawing.Those skilled in the art
It should be appreciated that following embodiments are only used for illustrating feasibility of the invention, not limit the invention.
The percentage composition being related in embodiment all refers to unless otherwise indicated (for example being explicitly indicated as percentage or ratio)
Mass percentage.
The screening of embodiment 1 and SiONFH related gene
1.1 according to Zhongshan Hospital Attached to Fudan Univ's Rheumatism Dept. inpatient on January 25,1 day to 2019 July in 2015
A perspective nested case-control study (Chinese Clinical Trial registration center number of registration: ChiCTR-OON-15005843,
Ethics official written reply code B2013-123 (2)), establish the Prospective Study queue of a SiONFH.The queue is included in because of various wind
Moist disease needs the patient using glucocorticoid, and hormone dosage need to reach conventional high dose (the i.e. equivalent bold and vigorous Buddhist nun of > 30mg/ days
Pine continues 30 days).Queue is included in qualifying patients 73, and follow-up crowd is Han nationality.The follow-up time limit 2 years, it is suitable to amount to 64 patients
Benefit completes follow-up, wherein 6 people of downright bad patient.It is matched according to Confounding Factors such as protopathy, gender, ages and chooses control 6 people of case,
The blood sample extracting genomic DNA of above-mentioned 12 patients simultaneously carries out full exon sequencing, filters out 9 difference sites, respectively
rs2305027、rs562038978、rs34433978、rs3833221、rs111896385、rs61758879、rs2708381、
Rs143471015, rs2277969, be related to 9 functional genes (COLEC12, FMN2, CYP2B6, CYP2A13, CYP2F1,
ARHGAP24,TAS2R46,SLFN12L,COL5A3).Variation type is related to nonsynonymous mutation, frameshift mutation, nonsense mutation and piece
Section deletes mistake.
1.2 correlation further to verify said gene and SiONFH, we establish a retrospective verifying queue,
Include 38, sample (wherein necrosis 23 compares 15).Above-mentioned 9 difference is detected by Sanger sequencing in verifying queue
Site, genotyping result find the OR value in tetra- sites rs34433978, rs2305027, rs3833221, rs143471015
(odds ratio) is respectively 19,15.7,9.1,0.2 (the equal < 0.1 of p), shows this 4 gene locis and SiONFH height phase
It closes.
1.3, when carrying out gene investigation, are included in the indication of patient: because a variety of causes is resulted in the need for using glucocorticoid
Class drug, daily dosage are greater than 30mg (equivalent prednisone), it is contemplated that using more than January, the age was between 18-65 years old patient.It needs
It is specifically intended that such as indulge in excessive drinking if the patient merges other specific caput femoris necrosis high risk factors, the hips such as fracture of neck of femur
Portion's primary disease, then no matter assessment result why, should all be disposed by high risk.
Biological sample leave and take and genome DNA extraction: using existing saliva acquisition method (such as saliva collection tube) or
Whole blood acquisition method (such as conventional venous blood collection) cdna collection group sample.- 80 DEG C of preservations are needed after sample collection, to unified extracting inspection
It surveys.QIAamp tissue DNA extraction agent box (QIAGEN company, the U.S.) or similar product can be used in the extracting of genomic DNA.
1.4 are directed to 4 gene locis, design PCR primer as shown in table 1 below.
Table 1, each gene loci amplimer
1.5 PCR amplification
PCR reaction system: forward primer, 10 μM, 1 μ L；Reverse primer, 10 μM, 1 μ L；2xTaq Master Mix, 10 μ
L；DNA sample, 1 μ g；Add ddH2O to 20 μ L.
PCR reaction condition: 95 DEG C of 5min；(95 DEG C of 30sec, Tm DEG C of 30sec, 72 DEG C of 1min) × 35 are recycled；72℃7min；
4 DEG C of heat preservations.
1.6 dideoxy-chain nucleic acid cessation methods (Sanger method) carry out DNA sequencing:
1) sample prepares: DNA sample being mixed with fluorescein, template in addition；
2) 3730 sequenator of ABI is run, electrophoresis is carried out to different size DNA in above-mentioned sample, so that by molecular size point
From and by inspection window；
3) when passing through window, fluorescein signal is detected, and collects and sequencing data is presented.
2 founding mathematical models of embodiment
2.1 based on embodiment 1 as a result, above-mentioned 4 gene locis layout score-sheet is summarized as shown in table 2 below
Statistical data.
The SiONFH clinical risk of each gene loci of table 2 scores
In table 2, by taking rs34433978 as an example, it is G base that " GG ", which represents two allele corresponding positions, and " G- " is represented
There is base deletion in one of allele, and " -- " represents two allele and base deletion occur；By taking rs2305027 as an example,
It is C base that " CC ", which represents two allele corresponding positions, and it is respectively C base and T base, " TT " that " CT ", which represents allele,
Representing two allele corresponding positions is T base；And so on.
2.2 combine clinical practice, and according to upper table 2, everybody comment split-phase is added as total score, and >=5 points are high risk, and≤4 points
For low-risk, increases and increase though risk scores.If scoring is the patient of high risk, it is proposed that use January, March, 6 after hormone
The moon regular follow-up hip magnetic resonance, as far as possible reduction hormone cumulative volume, or (if it is possible that) reduce hormone dosage to this hair
Bright is included in condition (the equivalent prednisone of 30mg/d) below.
Based on this, a kind of steroid femur head necrosis tumor susceptibility gene detection model is established, are as follows: the site F=
Rs2305027 scoring+site rs34433978 scoring+site rs3833221 scoring+site rs143471015 scores, wherein F
It is steroid femur head necrosis high risk as F >=5 for caput femoris necrosis (SiONFH) risk factor of steroids induction；When
It is steroid femur head necrosis low-risk when F≤4.
Obviously, which can continue to expand, and the scoring comprising other mutation types, this is those skilled in the art
It will be appreciated that.
At present there is no literature reported on there are close copy can Accurate Prediction SiONFH generation, several bases provided by the invention
The Combining diagnosis model or risk evaluation model of cause provide important references for prediction SiONFH.In perspective queue
Entirely because of necrosis rate 8.2%.Risk evaluation model is patient's necrosis rate 23.8% of high risk, is evaluated as patient's necrosis of low-risk
Rate 0.5% (p < 0.05).These results suggest that model accuracy with higher and Clinical significance of MG.
1 caput femoris necrosis classification confirmed result and above-mentioned mathematical model in conjunction with the embodiments, draws ROC curve, calculates bent
Area (AUG) under line, the AUC of four site Combining diagnosis models is up to 0.835 as the result is shown, sensitivity 0.87, specificity
0.80.As shown in Figure 1.
The clinical case of this mathematical model of the application of embodiment 3
Application Example verifies the prediction model established in embodiment 2.
One: SiONFH patient of case
38 years old female patients, Diagnosis of adult Still disease.Glucocorticoid 60mg/d is taken, patient's parting are as follows:
Rs2305027 CT, rs34433978--, rs3833221 CC, rs143471015 GG, SiONFH risk score is 7 points, point
Class is high risk.MRI shows bilateral femoral head necrosis after January.
Case two: non-SiONFH
53 years old female patients, diagnostic system lupus erythematosus.Glucocorticoid 40mg/d is taken, patient's parting are as follows:
Rs2305027 CC, rs34433978 GG, rs3833221--, rs143471015-G, SiONFH risk score is 1 point, classification
For low-risk.Have no caput femoris necrosis within follow-up 3 years.
Above embodiments verify technical solution of the present invention, it was demonstrated that the reliability of prediction model of the present invention,
Standard can be provided using glucocorticoid medicine patient and the prudent differentiation for using glucocorticoid medicine patient to be suitable
Really anticipation.It should be noted that those skilled in the art make various on this basis without prejudice under thought of the invention
Change or modification, equally should belong to the scope of the present invention.
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<213>artificial sequence ()
<400> 8
cgtggaaatg gcttagctgc 20
Claims (10)
1. a kind of steroid femur head necrosis tumor susceptibility gene detection model, are as follows: F=human genome site rs2305027 is commented
Point+site rs34433978 scoring+site rs3833221 scoring+site rs143471015 scoring, wherein F is hormonal femur
The downright bad risk factor of head, with steroid femur head necrosis risk correlation.
2. steroid femur head necrosis tumor susceptibility gene detection model as described in claim 1, which is characterized in that
The standards of grading of site rs2305027 are: rs2305027 CC i.e. two allele is C base in the site, note
It is 0 point；C base mutation of one of rs2305027 CT i.e. two allele in the site is T base, is denoted as 1 point；
C base of rs2305027 TT i.e. two allele in the site sports T base, is denoted as 2 points；
The standards of grading of site rs34433978 are: rs34433978 GG i.e. two allele is G base in the site,
It is denoted as 0 point；G base of one of rs34433978 G- i.e. two allele in the site is lost, and is denoted as 1 point；
G base of rs34433978-- i.e. two allele in the site is lost, and is denoted as 2 points；
The standards of grading of site rs3833221 are: the rs3833221-- i.e. two allele site is inserted into without base, is denoted as 0
Point；One of the rs3833221 C- i.e. two allele position inserts a C base, is denoted as 1 point；Rs3833221 CC is
Two allele sites insert a C base respectively, are denoted as 2 points；
The standards of grading of site rs143471015 are: rs143471015 GG i.e. two allele is G base in the site,
It is denoted as 2 points；The G base deletion of one of rs143471015 G- i.e. two allele in the position, is denoted as 1 point,
The G base of rs143471015-- i.e. two allele in the position lacks, and is denoted as 0 point.
3. steroid femur head necrosis tumor susceptibility gene detection model as claimed in claim 1 or 2, which is characterized in that when F >=5
When, it is steroid femur head necrosis high risk；It is steroid femur head necrosis low-risk as F≤4.
4. steroid femur head necrosis tumor susceptibility gene detection model as claimed in claim 1 or 2, which is characterized in that be entered
The message processing module of gene detection equipment or the message processing module for being entered blood testing equipment.
5. steroid femur head necrosis tumor susceptibility gene detection model as claimed in claim 1 or 2, which is characterized in that be entered
It intelligent medical system or is entered in the computer of doctor.
6. a kind of for detecting the kit of steroid femur head necrosis tumor susceptibility gene, which is characterized in that examined including being respectively used to
Survey human genome site rs2305027, rs34433978, rs3833221 and rs143471015 primer sequence, so as to
It is detected in carrying out amplification and subsequent sequencing to above-mentioned site.
7. kit as claimed in claim 6, which is characterized in that for detecting the primer pair of gene rs2305027 as forward direction
Primer SEQ ID NO:1 and reverse primer SEQ ID NO:2.
8. kit as claimed in claim 6, which is characterized in that for detecting the primer pair of gene rs34433978 as forward direction
Primer SEQ ID NO:3 and reverse primer SEQ ID NO:4.
9. kit as claimed in claim 6, which is characterized in that for detecting the primer pair of gene rs3833221 as forward direction
Primer SEQ ID NO:5 and reverse primer SEQ ID NO:6.
10. kit as claimed in claim 6, which is characterized in that the primer pair for detecting gene rs143471015 is positive
To primer SEQ ID NO:7 and reverse primer SEQ ID NO:8.
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| CN111690733A (en) * | 2020-06-22 | 2020-09-22 | 复旦大学附属中山医院 | Hormone-induced femoral head necrosis susceptibility gene panel |
| CN112410413A (en) * | 2020-09-24 | 2021-02-26 | 吉林大学 | Detection substance for ONFH (one-dimensional peptide binding) susceptibility related VDR (VDR), MMP2, MMP3 and MMP9 gene SNP and application |
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| CN113801931A (en) * | 2020-09-01 | 2021-12-17 | 吉林大学 | SNP detection substance related to ONFH risk genes CEBPA, PPAR gamma and CREBP and application thereof |
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