CN107090513A - GM2A genes diagnose the mark with prognosis as Male Osteoporosis - Google Patents
GM2A genes diagnose the mark with prognosis as Male Osteoporosis Download PDFInfo
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Abstract
The invention discloses the molecular labeling that GM2A genes could diagnose or predict prognosis as Male Osteoporosis.The present invention knows that GM2A genes content in the blood of Male Osteoporosis patient is significantly reduced than normal male by high-flux sequence and QPCR researchs, therefore GM2A genes can diagnose or predict the molecular labeling of prognosis as Male Osteoporosis.Examination of this method of osteoporosis available for healthy population, the generation of effective pre- anti-osteoporosis are characterized with the content of gene expression product in blood.
Description
Technical field
The invention belongs to biomedical sector, it is related to the molecular marker for diagnosing Male Osteoporosis.
Background technology
Osteoporosis is one group of osteopathy caused by many reasons, and bone tissue has normal calcification, and calcium salt is with matrix in just
Normal ratio, the metabolic bone disease with the characteristics of the reduction of unit volume inner bone tissues amount becomes.In most osteoporosises, bone tissue
Reduce caused by increasing mainly due to bone absorption.With skeleton pain, it is easy to fracture and is characterized.
Osteoporosis is classified as the disease for the second largest harm human health for being only second to angiocardiopathy by the World Health Organization
Disease.Promote the newest issue of foundation according to China's Healthy《2013 Chinese osteoporotic fracture preventing and treating blue books》It has been shown that, sclerotin is dredged
Pine fracture is a kind of fragility fractures, i.e., by low energy wound be generable bone in by minor trauma or daily routines
Folding.The common site of osteoporotic fracture is vertebra, hipbone and distal forearm.The people of more than 50 years old crowd of China about 69,440,000 suffers from
Osteoporosis, about 2.1 hundred million people's bone amount are relatively low.
In China, the degree of concern and the depth of investigation for osteoporosis in aged males are far from enough.But old man
Property osteoporosis harm and no less than women, the illness rate and case fatality rate of male's Hip Fracture are apparently higher than women.
Research shows that the hazards of Male Osteoporosis include both at home and abroad：
(1) age growth is an independent hazard factor of Male Osteoporosis.Shadow of the age for Bone m etabolism
Ring and potentially include relatively active to bone information, and osteogenic action is suppressed, and causes bone information to be more than bon e formation.At the same time,
With age, kidney l α hydroxylase activities decline, 1,25 (OH) of activity2D3Reduce, intestinal calcium absorption declines, blood calcium is reduced, and is drawn
SHPT is played, causes bone loss.
(2) body mass index (body mass index, BMI) BMI is to influence the one of osteoporosis in aged males illness rate
Individual important indicator.One important indicator of Male Osteoporosis illness rate.Studies have shown that BMI is less than 20-25kg/m2People
Group, which suffers from osteoporosis possibility, to increase, and is more than 25kg/m with BMI2Crowd compare, BMI is 15-20kg/m2Trouble
The possibility of person's marrow fracture can increase.In addition studies have found that, BMI often increases 1kg/m2Measure neck of femur, marrow and lumbar vertebrae
BMD increase 0.1SD, 0.11SD and 0.09SD respectively.BMI increases can cause the mechanical load of bone to increase, and stimulate bone shape
Into and reduce bone information.
(3) endocrine hormone influence androgen levels reduction be osteoporosis in aged males a significant risk because
Element.There is androgen receptor on osteocyte, androgen directly acts on these acceptors to promote Gegenbaur's cell to strengthen bone.It is old
The gonad index of one hypophysis of male's hypothalamus one lowers, hypo-orchidia, and testosterone levels decline, and sex hormone binding globulin
Level is raised, and is caused the osteogenic action of Gegenbaur's cell to weaken, is caused bone density to decline.
(4) inherent cause inherent cause occupies critical role in the morbidity of Male Osteoporosis.Current osteoporosis
Genetic research mainly includes Population Genetics and the broad aspect of molecular genetics two, respectively for ethnic group, family and gene pleiomorphism
Studied.The most significant determinant of Peak Bone Mass is inherent cause, and influence may account for 60%-80%.
(5) calcium agent and vitamin D in the composition proportioning especially food of trophic level patient chronic dietary custom and food
Morbidity of the content to osteoporosis also play certain effect.Meta analysis shows supplement the crowd of calcium and vitamine D3,
BMD has increased, the risk reduction fractured.Calcium agent can reduce bone amount loss, prevent vertebral fracture, and non-to preventing
Vertebral fracture may also be helpful.
In addition, habits and customs include the important risk factor that motion, smoking, heavy drinking are also Male Osteoporosis.
Diagnosis of osteoporosis it may first have to carry out the measure of bone density.Bone density is actually the health for representing bone
Degree, or the degree of aging of bone is represented on the contrary.Conventional detection means includes：
X ray photograph method：Usage history is earliest, but due to there is radioactivity, its test result is not energetic, is gradually taken
Generation.
Single photon analyzer：Expense is low, conveniently, amount of radiation it is small and safe.But because that can not survey bone of body and can not distinguish
Cortex bone, cancellous bone, soft tissue etc. and be restricted, be gradually substituted.
Quantitative Ultrasound Methods：It is cheap, conveniently and "dead" damage.It is adapted to the bone-shaped state census operations of each age group crowd.
Bone density can not only be detected, moreover it is possible to understand bone quality can reactive bone micro-structural and elasticity.But it is currently only used for calcaneum, shin
Bone and phalanges, it is impossible to carry out the detection of whole body bone.
Dual energy X-ray absorptiometry：It is current optimal determination method, is the goldstandard of Diagnosis of osteoporosis, it is logical both at home and abroad
With, each position bone of whole body can be surveyed, soft tissue thickness can be also surveyed, but the number of devices is less and expensive, therefore for bone
Financial burden is big for the diagnosis of matter osteoporosis patient.Examining for a kind of high sensitiveness, degree of accuracy height and reasonable price is found for this
The method of disconnected osteoporosis is urgent problem to be solved.
In recent years, inherent cause has become one of most active research field in bone biology, there are some researches show
VitD receptor genotype (VDR) and vitamin D binding protein (DBP) are the candidate genes of the relatively early osteoporosis found, separately
Outside it has also been found that ESR1 genes, ESR2 genes, CYP19A1 genes, CYP17A1 genes, ESRRA genes and ESRRG genes and sclerotin
The morbidity of osteoporosis has correlation.In addition, there are a considerable amount of patent applications to be related to the gene diagnosis of osteoporosis,
Such as Application No.：201610272604.0、201610922798.4、201510628024.6、201510628081.4、
201510725408.X、201510628042.4、201610530383.2、201510629348.1、201510627056.4、
201510627060.0,201610555353.7 patent.It can be seen that becoming bone from now on using gene come Diagnosis of osteoporosis
The development trend of matter osteoporosis early diagnosis.
The content of the invention
It is used to diagnose Male Osteoporosis or prediction Male Osteoporosis it is an object of the invention to provide one kind
The biomarker of prognosis.The present invention demonstrates GM2A genes in Male Osteoporosis using high-flux sequence and QPCR experiments
Expression in the blood of patient is substantially less than normal male, therefore can regard GM2A genes as diagnosis Male Osteoporosis
Or the biomarker of prediction Male Osteoporosis prognosis.
According to an aspect of the present invention, the invention provides detection GM2A (GM2ganglioside activator,
GM2 gangliosides activity factor) gene expression reagent preparing Male Osteoporosis auxiliary diagnosis or prediction prognosis production
Application in product.
Further, the reagent includes the reagent of detection GM2A mrna expressions, and/or detection GM2A albumen tables
Up to horizontal reagent.
The reagent of detection GM2A mrna expressions is such as PCR, such as Southern hybridization, Northern hybridization, point
Hybridization, FISH (FISH), DNA microarray, ASO methods, high-flux sequence platform etc. are qualitatively, quantitatively or semidefinite
Any reagent used in the method for amount ground detection mrna expression.
Further, the reagent of the detection GM2A mrna expressions includes the primer used in QPCR, and/or visits
Pin.
In specific embodiments of the present invention, the reagent of the detection GM2A mrna expressions is included in QPCR
The primer used, the primer sequence is as shown in SEQ ID NO.1 and SEQ ID NO.2.
Primer recited above can be prepared by chemical synthesis, using those skilled in the art will know that method refer to
Given information is prepared to be suitably designed by chemical synthesis.
Foregoing probe can be prepared by chemical synthesis, by using those skilled in the art will know that method
Appropriately design, and prepared by chemical synthesis with reference to Given information, or the phase can be contained by being prepared from biomaterial
The gene of nucleotide sequence is hoped, and expands it to prepare using the primer for expecting nucleotide sequence designed for amplification.
Detection GM2A protein expression levels reagent be as ELISA, radioimmunoassay, immunohistochemical method,
Western blot etc. can detect any reagent used in the method for protein expression level.
The present invention can be used for the reagent of detection GM2A protein expression levels to include the antibody for GM2A albumen, the antibody
Can be monoclonal antibody, polyclonal antibody, multi-specificity antibody (such as bispecific antibody).
The sample of the foregoing Product checking of the invention is not particularly limited, as long as it is suitable to the survey of the present invention
It is fixed；For example, it can include tissue, blood, blood plasma, serum, lymph, urine, serous cavity liquid, spinal fluid, synovia, aqueous humor,
Tear, saliva or its fraction or treated material.In specific embodiments of the present invention, the sample is from tested
The blood of person.
According to another aspect of the present invention, it is used for Male Osteoporosis auxiliary diagnosis or pre- present invention also offers one kind
The product of prognosis is surveyed, the product includes the reagent of foregoing detection GM2A gene expressions.
Further, the product before the present invention includes but is not limited to chip, kit, test paper or high-flux sequence
Platform；High-flux sequence platform is a kind of instrument of special diagnosis Male Osteoporosis, with high throughput sequencing technologies
Development, will turn into the structure of the gene expression profile of a people and very easily work.By contrasting Disease and normal person
The gene expression profile of group, the exception for easily analyzing which gene is related to disease.Therefore, GM2A is known in high-flux sequence
The exception of the gene purposes for falling within GM2A genes related to Male Osteoporosis, equally protection scope of the present invention it
It is interior.
Present invention also offers a kind of Male Osteoporosis auxiliary diagnosis or the method for predicting prognosis, methods described includes
Following steps：
(1) sample that male subject contains GM2A gene expression products is obtained；
(2) expression of GM2A genes or albumen in sample is detected；
(3) the GM2A genes measured or the expression of albumen are associated with the whether ill of subject.
(4) compared with normal control, if the expression reduction of GM2A genes or albumen, the male subject is examined
Break and be confirmed as poor prognosis for osteoporosis, or osteoporosis patient.
In the context of the present invention, " diagnosis Male Osteoporosis " had both included judging whether male subject has been suffered from
There is Male Osteoporosis, also include judging that male subject whether there is the risk with Male Osteoporosis.
Prediction prognosis refers to the process of prediction status of patient or result, is not meant to predict with 100% degree of accuracy
The process or result of status of patient.Prediction prognosis refers to whether the possibility for determining some processes or result increases, and simultaneously unexpectedly
Taste to be compared to determine the possibility for occurring some processes or result by situation about with some processes or result not occurring.Such as this
For invention, in the present invention in the patient of the level reduction of GM2A genes or GM2A albumen, compared with not showing the people of this feature,
More likely it was observed that particular procedure or result.
As used herein, term " antibody ", it is intended that be specifically bound to specific antigen or with specific antigen interact
Any antigen-binding molecule or molecular complex, it includes at least one complementary determining region (CDR).Term " antibody " includes containing
There are 4 polypeptide chains (that is, 2 weight (H) chains being connected with each other by cystine linkage and 2 light (L) chain) and its polymer is (for example
IgM).Each heavy chain contains weight chain variable district and heavy chain constant region.Heavy chain constant region contain 3 domain Cs H1, CH2 and
CH3.Each light chain contains light chain variable district and constant region of light chain.Constant region of light chain contains a domain (CL1).VH and VL
Area can be further subdivided into hypervariable region (being referred to as complementary determining region (CDR)), wherein being studded with more conservative referred to as framework region (FR)
Region.Each VH and VL is constituted by three CDR and four FR, is arranged in the following order from aminoterminal to c-terminus：FR1、
CDR1, FR2, CDR2, FR3, CDR3 and FR4.
As used herein, term " antibody " also includes the antigen-binding fragment of whole antibody molecule.The antigen binding of antibody
Fragment can utilize any suitable standard technique, for example proteolytic digestion or be related to encoding antibody variable and optionally it is constant
The recombination renovation technique of the DNA in area manipulation and expression, from for example complete antibody derivative molecule.Such DNA is
It is knowing and/or be easy to obtain from such as commercial source, DNA library (including such as bacteriophage-antibody library), or it can be synthesized.
The DNA can be sequenced and chemically or Protocols in Molecular Biology is manipulated, so that for example will be one or more variable
Area and/or constant region are arranged in suitable configuration, or introduce codon, produce cysteine residues, modification, addition or delete ammonia
Base acid etc..
The non-limiting examples of antigen-binding fragment include：(i) Fab fragments；(ii) fragments of F (ab') 2；(iii) Fd pieces
Section；(iv) Fv fragments；(v) scFv (scFv) molecule；(vi) dAb fragments；And (vii) by analog antibody hypervariable region (for example
Isolated complementary determining region (CDR), such as CDR3 peptides) amino acid residue composition minimum recognition unit or restricted type FR3-
CDR3-FR4 peptides.Antigen-binding fragment also includes the molecule of other engineering, such as domain-specific antibody, single domain antibody, knot
The antibody of structure domain missing, chimeric antibody, the antibody of CDR transplanting, double antibody, three antibody, four antibody, miniantibody, nano antibody (example
Such as monovalent nano antibody, divalence nano antibody), little module immune drug
(smallmodularimmunopharmaceuticals) (SMIP), and the variable IgNAR domains of shark.
" monoclonal antibody " be by monoclonal B- lymphocytes or will coding single antibody (or its antigen binding by
Fragment) antibody light chain and weight chain variable district the antibody that is produced to cell therein or its offspring of nucleic acid transfection.Monoclonal resists
Body is produced by method known to those skilled in the art, such as by merging manufacture from myeloma cell and immune spleen cell
Hybrid antibody forms cell to carry out.These fused cells and their offspring are referred to as " hybridoma ".
" multi-specificity antibody " can be have to the different epitopes of a target polypeptide specificity or may containing pair
More than one target polypeptide has specific antigen-binding domain.See, for example, Tutt etc., 1991, J.Immunol.147:60-
69；Kufer etc., 2004, TrendsBiotechnol.22:238-244.Antibody or its fragment can functionally link (example
Such as, by chemical coupling, genetic fusion, noncovalent associations or other modes) to one or more other molecular entities, it is such as another
Individual antibody or antibody fragment, to produce bispecific or multi-specificity antibody with second of binding specificity.
Brief description of the drawings
Fig. 1 displays detect that expression of the GM2A genes in Male Osteoporosis patient and normal male is poor using QPCR
It is different.
Specific embodiment
The present invention is further detailed explanation with reference to the accompanying drawings and examples.Following examples are merely to illustrate this
Invention rather than limitation the scope of the present invention.The experimental method of unreceipted actual conditions in embodiment, generally according to conventional strip
Part, such as Sambrook et al., molecular cloning：Laboratory manual (New York:Cold Spring HarborLaboratory
Press, 1989) described in condition, or according to the condition proposed by manufacturer.
Embodiment 1 screens the difference expression gene of Male Osteoporosis patient and normal male
1st, research object：
Bone density is detected, osteoporosis male patient 3 is selected, the age is respectively 60 years old, 82 years old, 68 years old；It is normal right
According to group male 2, the age is respectively 71 years old, 69 years old.
The inclusive criteria of patients with osteoporosis：(1) diagnosis of osteoporosis standard person, reference are met《Chinese's sclerotin
Osteoporosis suggestion diagnostic criteria (the second original text)；(2) the equal informed consent of patient.
Exclusion standard：Diabetes, serious liver and kidney disease, thyroid disease, endocrine system disease cause the elderly men of disease；
Took the elderly men for the treatment of osteoporosis medicine, such as Allan sodium phosphate (discontinuity is taken except low dose of calcium agent).
2nd, blood Total RNAs extraction
(1) pre-treatment
Fresh blood (peripheral blood) is directly taken, 3 times of volume erythrocyte cracked liquids are added, room temperature places 10 points after mixing
Clock, 10,000rpm centrifugations 1 minute.Thoroughly inhale and abandon supernatant, collect leukocyte cell pellet.Per the leucocyte of 100-200 μ l blood collections
Precipitation adds 1ml TRIzol.
(2) it is layered
A. sample is added after TRIzol, and room temperature places 5min, sample is fully cracked.
B. 200 μ l chloroforms are added per 1ml TRIzol, room temperature, which places 3-5min, after acutely vibration is mixed makes its natural split-phase.
(3) RNA precipitate
A.4 DEG C 12,000rpm centrifuges 10-15min.Sample can be divided into three layers：The organic phase of yellow, intermediate layer and colourless
Aqueous phase, RNA mainly in aqueous phase, aqueous phase (can generally draw 550 μ l) is transferred in new pipe.
B. isometric ice-cold isopropanol is added in supernatant, room temperature places 10-20min.4 DEG C of 12,000rpm centrifugations
10min, abandons supernatant, RNA precipitate is in ttom of pipe.
(4) RNA is rinsed
The ethanol of 1ml 75% (being prepared with RNase-free water) is added in a.RNA precipitations, centrifuge tube is gently vibrated, it is heavy to suspend
Form sediment.The ethanol of 1ml 75% is added per 1ml TRIzol.
B.4 DEG C 5,000-8,000rpm centrifugation 1-2min, abandon supernatant；Of short duration quick centrifugation, is carefully inhaled with pipettor and abandoned
Clearly, room temperature, which is placed, dries precipitation in 1-2 minutes.
(5) RNA is dissolved
50-100 μ l RNase-free water is added in precipitation, tube wall is flicked, fully to dissolve RNA, -70 DEG C of preservations.
3rd, the quality analysis of RNA sample
Carried RNA concentration and purity are detected using Nanodrop2000, agarose gel electrophoresis detection RNA is complete
Whole property, Agilent2100 determines RIN values.Single requirement for construction data base RNA total amounts 5 μ g, concentration >=200ng/ μ L, OD260/280 between
Between 1.8~2.2.
4th, fragmentation RNA
Illumina platforms are sequenced for short sequence fragment, and mRNA average lengths may reach several kb, it is therefore desirable to
It is interrupted at random.Using metal ion, can by RNA random fractures into 200bp or so small fragment.
5th, reversion synthesis cDNA
In the presence of reverse transcriptase, using random primer, by template of mRNA, one chain cDNA of reversion synthesis, carries out two chains
During synthesis, dTTP is replaced with dUTP in dNTPs reagents, base in the chains of cDNA second is included A/U/C/G.
6th, adaptor is connected
The cDNA structures of double-strand are cohesive end, add End Repair Mix and are mended into flat end, then at 3 ' ends
End is plus an A base, the joint for connecting Y-shaped.
7th, the chains of UNG enzymic digestions cDNA bis-
Before PCR amplifications, the chains of cDNA second are digested with UNG enzymes, so that only including the chains of cDNA first in library.
8th, machine is sequenced on Illumina x-ten
Illumina x-ten microarray datasets, carry out 2*150bp sequencings.
9th, bioinformatic analysis
It is as follows that sequencing data obtains later rawdata analyses process：
(1) trim is carried out to the 5 ' of reads and 3 ' sections with cutadapt, trim falls quality<20 base, and delete N
Reads more than 10%；
(2) tophat is compared onto reference gene group.Reference gene group version used be GRCh38.p7, fasta and
Gff file downloads are from NCBI；
(3) expression quantity of cuffquant quantification of mrna and normalization output；
(4) differential expression of the control group with disease group mRNA is compared with DEGseq bags under R environment.Significant difference mRNA is sieved
Select condition：p-value<0.05.
10th, result
Difference expression gene 3296 is obtained with above standard screening, wherein the gene of up-regulated expression 1428, under expression
The gene of tune has 1868.
The QPCR experimental verification Male Osteoporosis patients of embodiment 2 and the difference expression gene of normal male
The GM2A genes that selection example 1 is filtered out carry out large sample checking.
1st, research object：
Bone density is surveyed, selection osteoporosis male patient 30, the age is 60-82 Sui；Normal bone density control group man
Property 30, the age be 60-82 Sui.
The inclusive criteria of patients with osteoporosis：(1) diagnosis of osteoporosis standard person, reference are met《Chinese's sclerotin
Osteoporosis suggestion diagnostic criteria (the second original text)；(2) the equal informed consent of patient.
Exclusion standard：Diabetes, serious liver and kidney disease, thyroid disease, endocrine system disease cause the elderly men of disease；
Took the elderly men for the treatment of osteoporosis medicine, such as Allan sodium phosphate (discontinuity is taken except low dose of calcium agent).
2nd, blood Total RNAs extraction
(1) pre-treatment
Fresh blood (peripheral blood) is directly taken, 3 times of volume erythrocyte cracked liquids are added, room temperature places 10 points after mixing
Clock, 10,000rpm centrifugations 1 minute.Thoroughly inhale and abandon supernatant, collect leukocyte cell pellet.Per the leucocyte of 100-200 μ l blood collections
Precipitation adds 1ml TRIzol.
(2) it is layered
A. sample is added after TRIzol, and room temperature places 5min, sample is fully cracked.
B. 200 μ l chloroforms are added per 1ml TRIzol, room temperature, which places 3-5min, after acutely vibration is mixed makes its natural split-phase.
(3) RNA precipitate
A.4 DEG C 12,000rpm centrifuges 10-15min.Sample can be divided into three layers：The organic phase of yellow, intermediate layer and colourless
Aqueous phase, RNA mainly in aqueous phase, aqueous phase (can generally draw 550 μ l) is transferred in new pipe.
B. isometric ice-cold isopropanol is added in supernatant, room temperature places 10-20min.4 DEG C of 12,000rpm centrifugations
10min, abandons supernatant, RNA precipitate is in ttom of pipe.
(4) RNA is rinsed
The ethanol of 1ml 75% (being prepared with RNase-free water) is added in a.RNA precipitations, centrifuge tube is gently vibrated, it is heavy to suspend
Form sediment.The ethanol of 1ml 75% is added per 1ml TRIzol.
B.4 DEG C 5,000-8,000rpm centrifugation 1-2min, abandon supernatant；Of short duration quick centrifugation, is carefully inhaled with pipettor and abandoned
Clearly, room temperature, which is placed, dries precipitation in 1-2 minutes.
(5) RNA is dissolved
50-100 μ l RNase-free water is added in precipitation, tube wall is flicked, fully to dissolve RNA, -70 DEG C of preservations.
3rd, reverse transcription
RNA reverse transcription is carried out using the Reverse Transcriptase kit of TAKARA companies.
4、QPCR
(1) design of primers
QPCR amplimers are designed according to the coded sequence of GM2A genes in Genbank and β-actin genes, given birth to by Shanghai
Work biotechnology Services Co., Ltd synthesizes.Specific primer sequence is as follows：
GM2A genes：
Forward primer is 5 '-CTCTGAAGGTGGATTTAGTT-3 ' (SEQ ID NO.3)；
Reverse primer is 5 '-CTGCCAATGTAGTCTGTG-3 ' (SEQ ID NO.4),
β-actin genes：
Forward primer is 5 '-GCAGGTCATCACCATCGG-3 ' (SEQ ID NO.5)；
Reverse primer is 5 '-GCTGTCACCTTCACCGTTC-3 ' (SEQ ID NO.6).
(2) PCR reaction systems are prepared according to table 1：
Wherein, SYBR Green PCRs system is purchased from Invitrogen companies.
The PCR reaction systems of table 1
| Reagent | Volume |
| Forward primer | 1μl |
| Reverse primer | 1μl |
| SYBR Green PCRs | 12.5μl |
| System template | 2μl |
| Deionized water | Supply 25 μ l |
(3) PCR reaction conditions：95 DEG C of 10min, (95 DEG C of 15s, 60 DEG C of 40s) * 45 circulations.Using SYBR Green as
Fluorescent marker, in the enterprising performing PCR reaction of Light Cycler quantitative real time PCR Instruments, is analyzed by melt curve analysis and electrophoresis is true
Determine purpose band, Δ Δ CT methods carry out relative quantification.
5th, result
As a result as shown in figure 1, compared with normal male, the mRNA water of GM2A genes in Male Osteoporosis blood samples of patients
Flat to be remarkably decreased, difference has statistical significance (P<0.05), as a result tested with RNA-seq.
The explanation of above-described embodiment is only intended to understand the method and its core concept of the present invention.It should be pointed out that for this
For the those of ordinary skill in field, under the premise without departing from the principles of the invention, some improve can also be carried out to the present invention
And modification, these are improved and modification will be also fallen into the protection domain of the claims in the present invention.
SEQUENCE LISTING
<110>Beijing Friendship Hospital Attached to Capital Medical Univ.
<120>GM2A genes diagnose the mark with prognosis as Male Osteoporosis
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 20
<212> DNA
<213>Artificial sequence
<400> 1
ctctgaaggt ggatttagtt 20
<210> 2
<211> 18
<212> DNA
<213>Artificial sequence
<400> 2
ctgccaatgt agtctgtg 18
<210> 3
<211> 18
<212> DNA
<213>Artificial sequence
<400> 3
gcaggtcatc accatcgg 18
<210> 4
<211> 19
<212> DNA
<213>Artificial sequence
<400> 4
gctgtcacct tcaccgttc 19
Claims (10)
1. detect reagent the answering in Male Osteoporosis auxiliary diagnosis or prediction prognosis product is prepared of GM2A gene expressions
With.
2. application according to claim 1, it is characterised in that the reagent includes detection GM2A mrna expressions
Reagent, and/or detection GM2A protein expression levels reagent.
3. application according to claim 1 or 2, it is characterised in that the examination of the detection GM2A mrna expressions
Agent includes the primer used in QPCR, and/or probe.
4. application according to claim 3, it is characterised in that the sequence of the primer such as SEQ ID NO.1 and SEQ ID
Shown in NO.2.
5. application according to claim 1 or 2, it is characterised in that the reagent bag of the detection GM2A protein expression levels
Include the antibody for GM2A albumen.
6. application according to claim 5, it is characterised in that the antibody includes monoclonal antibody, polyclonal antibody, many
Specific antibody.
7. application according to claim 1, it is characterised in that the sample of the Product checking is blood sample.
8. a kind of product for being used for Male Osteoporosis auxiliary diagnosis or predicting prognosis, it is characterised in that the product includes
The reagent of detection GM2A gene expressions described in claim 1.
9. product according to claim 8, it is characterised in that the product includes kit, chip or test paper.
10. product according to claim 8 or claim 9, it is characterised in that the sample of the Product checking is blood sample.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710502537.1A CN107090513A (en) | 2017-06-27 | 2017-06-27 | GM2A genes diagnose the mark with prognosis as Male Osteoporosis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710502537.1A CN107090513A (en) | 2017-06-27 | 2017-06-27 | GM2A genes diagnose the mark with prognosis as Male Osteoporosis |
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| CN107090513A true CN107090513A (en) | 2017-08-25 |
Family
ID=59641479
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710502537.1A Pending CN107090513A (en) | 2017-06-27 | 2017-06-27 | GM2A genes diagnose the mark with prognosis as Male Osteoporosis |
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| CN (1) | CN107090513A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108060222A (en) * | 2017-12-29 | 2018-05-22 | 北京泱深生物信息技术有限公司 | Application of the RBBP4 genes in clinical application |
| CN109055542A (en) * | 2018-09-27 | 2018-12-21 | 中国航天员科研训练中心 | Predict the method and DNA methylation marker of weightless bone loss lumbar spine bmd downside risk |
| WO2021082350A1 (en) * | 2019-10-30 | 2021-05-06 | 河北工业大学 | Tmem16a acting as osteoporosis marker and application thereof, osteoporosis diagnostic kit and drug |
| WO2024012583A1 (en) * | 2022-07-14 | 2024-01-18 | 浩峰生物科技股份有限公司 | Humanized fucosylated gm2ap antibody |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108060222A (en) * | 2017-12-29 | 2018-05-22 | 北京泱深生物信息技术有限公司 | Application of the RBBP4 genes in clinical application |
| CN109055542A (en) * | 2018-09-27 | 2018-12-21 | 中国航天员科研训练中心 | Predict the method and DNA methylation marker of weightless bone loss lumbar spine bmd downside risk |
| WO2021082350A1 (en) * | 2019-10-30 | 2021-05-06 | 河北工业大学 | Tmem16a acting as osteoporosis marker and application thereof, osteoporosis diagnostic kit and drug |
| WO2024012583A1 (en) * | 2022-07-14 | 2024-01-18 | 浩峰生物科技股份有限公司 | Humanized fucosylated gm2ap antibody |
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