CN107653311A - The SNP rs4883263 detecting system related to blood lipid level and related application - Google Patents
The SNP rs4883263 detecting system related to blood lipid level and related application Download PDFInfo
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Abstract
The invention provides a kind of SNP rs4883263 detecting system related to blood lipid level and its application；Specifically present invention determine that SNP rs4883263 is related to blood lipid level, provide the reagent material of the polymorphism in rs4883263 sites and/or instrument and equipment in sample of the detection from test individual and prepare the application in being used to assess the detecting system of blood lipid level and/or dyslipidemia onset risk, a kind of detecting system for assessing blood lipid level and/or dyslipidemia onset risk is additionally provided, it includes：Detect the reagent material and/or instrument and equipment of the polymorphism in rs4883263 sites in the sample from test individual；Wherein, rs4883263 and HDL C levels are significantly correlated, and rs4883263 sites C allele carrier has the horizontal and higher dyslipidemia onset risks of relatively low HDL C than noncarrier.
Description
Technical field
The present invention relates to the SNP rs4883263 detecting system related to blood lipid level and related application,
Specifically the present invention relates to the examination of the polymorphism in CD16 gene rs4883263 sites in sample of the detection from test individual
Agent material and/or instrument and equipment are being prepared for assessing in the detecting system of blood lipid level and/or dyslipidemia onset risk
Using further relating to a kind of detecting system for assessing blood lipid level and/or dyslipidemia onset risk.
Background technology
Numerous studies data shows that dyslipidemia is coronary heart disease, myocardial infarction, sudden cardiac death and cerebral arterial thrombosis
Independent hazard factor.It by accelerating systemic atherosclerosis, to body cause concealment, gradually, progressive, it is systemic and
Organic infringement.Studies have shown that crowd's blood cholesterol levels often raise 1%, incidence of coronary heart disease increase by 2%~3%.China
It is in notable trend, 2007~2008 years China national diabetes and metabolic disorder result of study that patients with dyslipidemia sum, which rises,
Prompting, more than the 20 years old crowd's cholesterol levels in China added 23.9% compared with 2002, and dyslipidemia has become China
The important public hygiene problem of resident.2011 annual Beijing's health show that Beijing is normal with population health status report data
Super half citizen dyslipidemia in the people is lived, wherein the dyslipidemia illness rate of 18~30 years old male has reached 58.5%.
Total plasma cholesterol (TC), LDL-C (LDL-C), HDL-C (HDL-C)
It is the target spot and target of the most important hazards of angiocardiopathy and Results with triglycerides (TG) concentration.Defend in the world
During raw tissue update prompting angiocardiopathy prevention and control benefit, contribution caused by the control of dyslipidemia hazards is maximum.
U.S.'s coronary heart disease death drop by half during 1980~2000 years, be attributed to the fact that carry out made by national cholesterol education completely
Contribution.
Most of dyslipidemias have complicated Etiologic Mechanism, be between multiple genes or multiple genes with environment because for a long time
The result of interaction.Genome-wide association study has been successfully determined multiple genetic locuses related to blood fat.It is however, several
All these sites are determined in the crowd of European descent, are lacked in asian population particularly Chinese population
Data.The difference of environmental exposure and genetic background between Chinese and European is huge, therefore needs identification China badly
The special blood fat variant sites of Chinese Han Population, it will help control the prevention of compatriots' hyperlipidemia, new drug development, diagnosis and individuation
Treat.
The content of the invention
It is a primary object of the present invention to determine the related to blood lipid level of asian population particularly gook's group specificity
Genetic locus, there is provided by detect correlated inheritance site assess blood lipid level or assess dyslipidemia onset risk side
Method, and the detecting system and related application for assessing blood lipid level and/or dyslipidemia onset risk are provided.
Inventor includes Chinese, Filipino, Chinese American, the Singapore nationality foreign citizen of Chinese origin, Singapore with East Asia crowd
Nationality Malaysian etc. is research object, analyzed and researched individual SNP (SNP) more than 110,000 and TC, LDL-C,
HDL-C, TG incidence relation, it is determined that mononucleotide polymorphism site rs4883263 is related to blood lipid level.rs4883263
Site is the genetic locus related to blood lipid level of gook's group specificity.The primary dcreening operation experimental stage data of the present invention
Show, the rs4883263 and HDL-C of CD163 gene regions are significantly correlated, up to full-length genome significance (P=5.24 ×
10-11)；In the repeated authentication stage, site P values still significantly (P=1.45 × 10-3), illustrate that the association is highly reliable；Will
Two stage sample combined analysis, P values reach 1.71 × 10-13.The present invention is further discovered that rs4883263 C allele
Frequency is that 0.69, C allele can cause HDL-C (HDL-C) reduction.
So as to by detecting the polymorphism in rs4883263 sites in the sample from test individual, can be used for assessment and treat
Individual blood lipid level particularly HDL-C (HDL-C) level is surveyed, may also be used for assessing test individual blood fat
Abnormal onset risk.
On the one hand, the invention provides the examination of the polymorphism in rs4883263 sites in sample of the detection from test individual
Agent material and/or instrument and equipment are being prepared for assessing in the detecting system of blood lipid level and/or dyslipidemia onset risk
Using.
According to specific embodiments of the present invention, in of the invention, blood lipid level includes HDL-C
(HDL-C)。
According to specific embodiments of the present invention, in of the invention, rs4883263 risk allele is C.
According to specific embodiments of the present invention, in of the invention, rs4883263 sites C allele carrier takes than non-
There is relatively low High-density Lipoprotein-cholesterol and/or higher dyslipidemia onset risk with person.Specifically,
Rs4883263 sites C allele carrier has relatively low High-density Lipoprotein-cholesterol than noncarrier, has
Higher plasma total cholesterol levels elevated risk, and/or with higher low-density lipoprotein cholesterol level rise wind
Danger.
According to specific embodiments of the present invention, in the present invention, the test individual is East Asia crowd, including Chinese,
Filipino, Chinese American, the Singapore nationality foreign citizen of Chinese origin, Singapore nationality Malaysian etc..During specific detection, the sample can come
From the blood of test individual, urine, saliva, gastric juice, hair or biopsy etc., preferably blood.
Possible technique any in this area can be used to detect mononucleotide of the present invention in DNA level, rna level
Pleomorphism site.Such as：Can use direct Sequencing method, by DNA direct Sequencings can directly disclose crt gene and
The sequence difference between mutator is carried, can be specifically traditional use business sequencing kit or automatic sequencer pair
DNA is directly sequenced, or develop in recent years pyrosequencing (Pyrosequencing), micro sequence (SNaPshot)
Deng.The method based on hybridization can also be used, specifically includes Taqman sonde methods, DNA chip method etc..It can also use and be based on
The method of primer extend, such as Matrix Assisted Laser Desorption ion flight time mass spectrum (MALDI-Tof-MS).Base can also be used
In the method for conformation, specifically such as RFLP (RFLP) analysis, single-strand conformation polymorphism (single-
Strand conformational polymorphism, SSCP) analyze, denaturing gradient gel electrophoresis (denaturing
Gradient gel electrophoresis, DGGE) analyze, Denaturing high performance liquid chromatography (denaturing high
Performance liquid chromatography, dHPLC) etc. analytical technology.High-resolution dissolving can also be used bent
Line analysis technology (HRM).In the specific implementation, those skilled in the art can select above-mentioned any according to actual conditions
Kind technology vitro detection mononucleotide polymorphism site of the present invention, can also be examined in vitro using the combination of multiple technologies
Survey the mononucleotide polymorphism site.
According to specific embodiments of the present invention, in of the invention, the reagent of the polymorphism in the detection rs4883263 sites
Material and/or instrument and equipment can be used in any feasible technology for detecting the mononucleotide polymorphism site
Reagent material and/or instrument and equipment etc..Such as：Reagent for direct sequencing；Or for PCR and limitation
Property the reagent that is combined of fragment length polymorphism analysis；Or the examination being combined for PCR with direct sequencing
Agent；Or the reagent being combined for PCR with direct sequencing；Or for following any SNP classifying methods
Reagent：Method based on hybridization, the method based on primer extend, the method based on conformation or high-resolution solubility curve point
Analysis technology etc..
On the other hand, present invention also offers a kind of detection system for assessing blood lipid level and/or dyslipidemia onset risk
System, it includes：The reagent material of the polymorphism in rs4883263 sites and/or instrument are set in sample of the detection from test individual
It is standby.
According to specific embodiments of the present invention, of the invention blood lipid level and/or the dyslipidemia onset risk assessed
Detecting system, it includes detection unit and assessment unit, wherein：
The detection unit includes the reagent material of the polymorphism in rs4883263 sites in sample of the detection from test individual
Material and/or instrument and equipment, the testing result of rs4883263 sites risk allele situation is carried for obtaining test individual；
The assessment unit includes being used for the processing unit for according to the testing result of detection unit assess processing；Its
In, rs4883263 sites C allele carrier than noncarrier have relatively low High-density Lipoprotein-cholesterol and/
Or dyslipidemia onset risk.
The detecting system for assessing blood lipid level and/or dyslipidemia onset risk of the present invention, can be virtual bench, only
The function of the detection unit and assessment unit can be realized.Described detection unit can include various detections
Reagent material and/or detecting instrument equipment etc.；Described data analysis unit can be any can realize to detection unit
Testing result is analyzed and processed and draws blood lipid level and/or computing instrument, the mould of dyslipidemia onset risk assessment situation
Block or virtual unit, such as can be in advance by various possible testing results and corresponding blood lipid level (including HDL-C
It is horizontal) and/or the corresponding data drawing list of dyslipidemia onset risk formulation, the testing result of detection unit is compareed into the data
Chart can draw blood lipid level and/or dyslipidemia onset risk assessment result.
Using the technology of the present invention, East Asia crowd's blood lipid level can be assessed by detecting rs4883263 loci polymorphisms
And/or dyslipidemia onset risk, the individuation health action scheme for research object is made, and will be helpful to state
The prevention of people's hyperlipidemia, new drug development, diagnosis and individualized treatment.
Brief description of the drawings
Fig. 1 shows rs4883263 meta analysis results.
Embodiment
In order to be more clearly understood that the present invention, the present invention is further described referring now to the following example and accompanying drawing.Embodiment
It is only used for explaining without limiting the invention in any way.The experimental method of unreceipted actual conditions is led to be affiliated in embodiment
Conventional method known to domain and normal condition, or according to the condition proposed by manufacturer.
Embodiment one
(primary dcreening operation stage and Qualify Phase) detects and analyzed respectively hereditary variation and TC, LDL- to the present invention in two stages
C, hereditary variation related to TG HDL-C.
Study population includes 13408 research objects of 47532 research objects of first stage and Qualify Phase, full-fledged research
Include East Asia crowd 61669 altogether.Each research obtains its research institution and the approval of local fact-finding organ Ethics Committee.It is all
The written signature informed consent form of participant.
First stage, extron group association study meta analyses are carried out to 47532 respondents of East Asia crowd.47532
Example research object, examined from Chinese ophthalmology research (CHES), China's Healthy and nutrition survey (CHNS), Port of Fangcheng men's health
Look into investigation (FAMHES), Guizhou Bijie diabetes B research (GBTDS), Hong Kong University's special item plan (HKU-TRS), lake
23 researchs of northern coronary heart disease research (HuCAD) and Chinese aged's nutrition and investigation of health conditions (NHAPC) etc..Crowd's base
This information is referring to table 1.Genotyping platform, genotype-phenotype analysis software etc. used in 23 independent blood lipid level researchs
Situation is shown in Table 2.Merge all samples and be associated analysis, after excluding singlet site final 110986 hereditary variation include pass
Connection analysis, the significance,statistical of research are set to P<4.5×10-7.Select the site significantly associated with East Asia crowd's blood lipid level
Repeated authentication is carried out in independent sample.
In the first stage, to HDL-C, LDL-C, TG and TC index measured in each queue all by age, age
Quadratic sum respectively study specific covariant be adjusted and be converted into average be 0 standard deviation be 1 standardized normal distribution to improve
Comparativity between different variables.Using RAREMETALWORKER or RVTESTS softwares respectively to analyzing each research sample
Middle hereditary variation and HDL-C, LDL-C, TC and TG incidence relation, and then carry out meta analyses using RAREMETALS softwares and close
And the result of each research.It is consistent with the first stage in the blood lipid level analytical plan of each research of Qualify Phase.Finally apply
METAL fixed effect inverse weight meta analyses merge two benches result.
Table 1, first stage Exon chip detection crowd's essential information
Abbreviation：CHES, Chinese ophthalmology research；CHNS, China's Healthy and nutrition survey；CLHNS, cebu longitudinal direction health and battalion
Support investigation；FAMHES, Port of Fangcheng men's health inspection investigation；GBTDS:Guizhou Bijie diabetes B research；
HKU-TRS, Hong Kong University's special item plan；HuCAD, Hubei coronary heart disease research；NHAPC, the Chinese aged
Nutrition and investigation of health conditions；PUUMA, Peking University's health science center and medical college of University of Michigan myocardial infarction are ground
Study carefully；SBCS, Shanghai breast cancer research；SCES, the research of Chinese eye section of Singapore；SiMES, Singapore's Malaysian's ophthalmology research；
SMHS, Shanghai men's health research；SP2, Singapore's perspective study project；SWHS, Shanghai women's health research；TUDR is beautiful
State's TaiWan, China people diabetic retinopathy research；TC, T-CHOL；LDL-C, LDL-C；HDL-C is high
Density lipoprotein-cholesterol；TG, triglyceride；BMI, body mass index.
Table 2, first stage research sample Exon chip detection technique platform and analysis software
In the present invention, the first stage have rated in 47532 respondents from East Asia crowd 110,000 SNPs with
TC, LDL-C, HDL-C and TG incidence relation.
The analysis result of first stage shows that the rs4883263 and HDL-C of CD163 gene regions are significantly correlated, up to full base
Because of group significance (P=5.24 × 10-11), meta analysis results are shown in Fig. 1.
Second stage, Qualify Phase.Select China's atherosclerosis study I and II (CAS, CAS- II), Beijing artery
Repeated authentication is carried out in atherosis research (BAS), brine sensitivity genetic epidemiology network (GenSalt) crowd, is wrapped altogether
Include 13408, sample.Crowd's essential information is referring to table 3.Select first stage and blood lipid level notable from these GWAS researchs
The site of association checks its genotyping result and verifies its correlation with blood lipid level.Five independent blood lipid level research institutes make
Situations such as Genotyping platform and genotype-phenotype analysis software, is shown in Table 4.
Table 3, repeated authentication stage crowd's essential information
Abbreviation：CAS, Chinese atherosclerosis study；BAS, Beijing atherosclerosis study；GenSalt, salt are quick
Perceptual genetic epidemiology network research；TC, T-CHOL；LDL-C, LDL-C；HDL-C, high density fat
Protein cholesterol；TG, triglyceride；BMI, body mass index.
Table 4, second stage research sample GWAS research genotyping techniques platform, analysis software
In repeated authentication, all sample vacuum blood taking needle venipunctures are gathered with venous blood overnight, measure is consolidated comprising total courage
Alcohol, HDL-C, TG are horizontal.Blood sample determines facing in Beijing Fu Wai Hospital, Chinese Academy of Medical Sciences population genetic research department
The experiment of bed inspection center is completed.The laboratory participates in the lipid standardization progam of CDC of the U.S..Hitachi
7060 automatic clinical chemistry analyzers determine the solid HDL-C of total courage and triglycerides.It is dense that LDL-C is calculated using Friedewald equations
Degree.
It is consistent with the first stage to change blood lipid level standard normal in Qualify Phase, using linear regression model (LRM), additivity
The correlation of model analysis blood lipid level (continuous variable) and selected site parting, and correct the age, the age square, sex and
Body mass index variable.
In the repeated authentication stage, rs4883263 sites P values still significantly (P=1.45 × 10-3), illustrate that the association can
It is strong by property.
Merge two benches result using METAL fixed effect inverse weight meta analyses.
Table 5 lists the rs4883263 and blood fat relevance result data that the present invention studies.
Table 5, rs4883263 and blood fat association results
Primary dcreening operation stage data shows that the rs4883263 and HDL-C of CD163 gene regions are significantly correlated, up to full genome
Group significance (P=5.24 × 10-11), meta analysis results are shown in Fig. 1；In the repeated authentication stage, site P values are still
Significantly (P=1.45 × 10-3), illustrate that the association is highly reliable；By two stage sample combined analysis, P values reach 1.71 ×
10-13, as shown in Table 5, rs4883263 C gene frequencies are 0.69, HDL-C can be caused to reduce.
Claims (10)
1. the reagent material of the polymorphism in rs4883263 sites and/or instrument and equipment exist in sample of the detection from test individual
Prepare for assess blood lipid level and/or dyslipidemia onset risk detecting system in application.
2. application according to claim 1, wherein, it is horizontal that blood lipid level includes HDL-C (HDL-C).
3. application according to claim 1, wherein, rs4883263 risk allele is C.
4. application according to claim 1, wherein, rs4883263 sites C allele carrier has than noncarrier
Higher dyslipidemia onset risk.
5. application according to claim 1, wherein, rs4883263 sites C allele carrier has than noncarrier
Relatively low High-density Lipoprotein-cholesterol.
6. application according to claim 1, wherein, the test individual is East Asia crowd.
7. application according to claim 1, wherein, blood of the sample from test individual, urine, saliva, gastric juice, head
Hair or biopsy, preferably blood.
8. a kind of detecting system for assessing blood lipid level and/or dyslipidemia onset risk, it includes：Detection comes from test individual
Sample in rs4883263 sites polymorphism reagent material and/or instrument and equipment.
9. detecting system according to claim 8, it includes detection unit and assessment unit, wherein：
The detection unit includes the reagent material of the polymorphism in rs4883263 sites in sample of the detection from test individual
And/or instrument and equipment, for obtaining the testing result of test individual carrying rs4883263 sites risk allele situation；
The assessment unit includes being used for the processing unit for according to the testing result of detection unit assess processing；Wherein,
Rs4883263 sites C allele carrier than noncarrier have relatively low High-density Lipoprotein-cholesterol and/or compared with
High dyslipidemia onset risk.
10. detecting system according to claim 8 or claim 9, wherein, in the sample of the detection from test individual
The reagent material and/or instrument and equipment of the polymorphism in rs4883263 sites include：Reagent for direct sequencing；Or it is used for
The reagent that PCR is combined with restriction fragment length polymorphism analysis；Or for PCR and directly
Connect the reagent that PCR sequencing PCR is combined；Or the reagent being combined for PCR with direct sequencing；Or for following
The reagent of any SNP classifying methods：Method based on hybridization, the method based on primer extend, the method based on conformation or height
Resolution ratio solubility curve analytical technology.
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| CN113604560A (en) * | 2021-08-25 | 2021-11-05 | 华中科技大学同济医学院附属协和医院 | SNP (single nucleotide polymorphism) related to blood fat and application thereof |
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