CN102021238A - ADRB1 (adrenergic, beta-1) gene SNP (Single Nucleotide Polymorphism) detection specificity primer and liquid phase chip - Google Patents
ADRB1 (adrenergic, beta-1) gene SNP (Single Nucleotide Polymorphism) detection specificity primer and liquid phase chip Download PDFInfo
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Abstract
The invention discloses an ADRB1 (adrenergic, beta-1) gene SNP (Single Nucleotide Polymorphism) detection specificity primer and a liquid phase chip. The liquid phase chip mainly comprises an ASPE primer, microspheres and an amplification primer, wherein the ASPE (Allele Specific Primer Extension) primer comprises a tag sequence of 5' end and a specific primer of 3' end aiming at an SNP site, and the specific primer is SEQ ID NO.5 and SEQ ID NO.6 aiming at the Al45G SNP site and/or SEQ ID NO.7 and SEQ ID NO.8 aiming at the C1165G SNP site; the microspheres are respectively coated with a specific anti-tag sequence and have different color codes, and the anti-tag sequence is selected from SEQ ID NO.9 to SEQ ID NO.12; and the amplification primer is used for amplifying an ADRB1 gene target sequence with Al45G and/or C1165G SDP site. The coincidence rate of the results of the detection liquid phase chip and the sequencing method is as high as 100%. The specificity primer has good specificity and can accurately distinguish various types of genotypes.
Description
Technical field
The invention belongs to biology field, relate to medical science and biotechnology, concrete a kind of ADRB1 gene test Auele Specific Primer and the liquid-phase chip of relating to.
Background technology
Adrenergic receptor gene (adrenergic receptor, ADRB) divide 3 kinds of hypotypes, be respectively ADRB1 (β 1-adrenergicreceptor), ADRB2 and ADRB3, the adrenergic receptor of its coding is one of receptor family member with guanosint glycosidoprotein (G albumen) coupling connection, is distributed widely in adipocyte.Adrenergic receptor combines back activated adenyl cyclase (AC) with catecholamine increase cyclic monophosphate (cAMP) generation, further activated protein kinase A, by acting on multiple lipid metabolism involved enzyme class, ionic channel and transcription factor, promote steatolysis.
Wherein, the ADRB1 gene is positioned at human chromosome 10q24-q26 zone, and the β1Shen Shangxiansunengshouti of its coding is the important adrenoceptor of a class that is present in the myocardial cell, is made up of 477 amino-acid residues, belongs to the g protein coupled receptor superfamily.There are 2 important single nucleotide polymorphism (SNP) in the ADRB1 gene, one is A145G (rs1801252), 49 Serines of coding are replaced by glycine (Ser → Gly), another site G1165C (rs1801253), corresponding the 389th amino acids becomes arginine (Gly → Arg) by glycine.The Gly389Arg variation is positioned at cell C-terminal Gs albumen coupling position, can strengthen adenylate cyclase activity, and the mutation frequency in Chinese is about 70%.The correlation research of American scholar Johnson etc. finds, it almost is 3 times of other different genotype persons that 389Arg homozygote patient takes behind the metoprolol drop-out value of diastolic pressure in the daytime.Sofowora etc. have also obtained similar results to 34 healthy volunteers' research, and behind the oral 25mg atenolol USP 23,389Arg homozygote volunteer's the vein systolic pressure and the decline of mean arterial pressure all are significantly higher than 389Gly homozygote person.
In addition, experimentation on animals confirms all that with the cohort study 2 polymorphism Gly389Arg, Ser49Gly of ADRB1 gene are with fat relevant.Carrying the allelic U.S. of 389Arg Caucasia women has higher BMI and body weight, and Sweden women's the BMI that carries the Gly389Arg transgenation is than the women's height that carries the Ser49Gly transgenation.The Swede of carrying Gly49-Gly389 combination gene type compares BMI rising 0.7kg/m with carrying other genotypic Swedes
2
The product that detects ADRB1 at present generally is based on the RT-PCR of round pcr, quantitative fluorescent PCR, traditional solid phase chip technology, direct order-checking, PCR-RFLP etc., this technology exists sensitivity low, the shortcoming that sample easily pollutes, false positive rate is high, owing to detect the limitation of flux, can not satisfy the needs of practical application simultaneously.
Summary of the invention
One of purpose of the present invention provides the ADRB1 gene SNP detection liquid-phase chip.This liquid-phase chip can be used for detecting wild-type and the mutant of two kinds of common genotype A145G of ADRB1 gene (rs1801252) and C1165G (rs1801253).
The technical scheme that realizes above-mentioned purpose is:
A kind of ADRB1 gene SNP detection liquid-phase chip mainly includes:
(A) at the SNP site of ADRB1 gene, the ASPE primer of She Ji wild-type and mutant is right respectively: every kind of ASPE primer is made up of at the Auele Specific Primer in SNP site the tag sequence and the 3 ' end of 5 ' end, and described Auele Specific Primer is: at the SEQ ID NO.5 in A145G SNP site and SEQ ID NO.6 and/or at the SEQ ID NO.7 and the SEQ ID NO.8 in C1165G SNP site; Described tag sequence is selected from SEQ ID NO.1～SEQ ID NO.4;
(B) be coated with microballoon special anti-tag sequence, that have the different colours coding respectively, described anti-tag sequence is selected from the sequence among SEQID NO.9～SEQ ID NO.12, and described anti-tag sequence can be correspondingly with (A) in selected tag sequence complementary pairing;
(C) be used to amplify the amplimer of the ADRB1 gene target sequence in SNP site with A145G and/or C1165G.
Preferably, described amplimer is: at the SEQ ID NO.13 in A145G SNP site and SEQ ID NO.14 and/or at the SEQ ID NO.15 and the SEQ ID NO.16 in C1165G SNP site.
Preferably, also be provided with the spacerarm sequence in the middle of described anti-tag sequence is connected with microballoon; Described spacerarm sequence is 5-10 T.
Another object of the present invention provides the Auele Specific Primer that a kind of ADRB1 of being used for gene SNP detects.
Concrete technical scheme is as follows:
Mainly include: at the SEQ ID NO.5 in A145G SNP site and SEQ ID NO.6 and/or at the SEQ ID NO.7 and the SEQ ID NO.8 in C1165G SNP site.
Major advantage of the present invention is:
1. the result of detection liquid-phase chip provided by the present invention and sequencing coincide rate up to 100%.Prepared ADRB1 gene SNP detection liquid-phase chip has extraordinary signal-noise ratio, and there is not cross reaction between designed probe and the anti-tag sequence basically, choosing of tag sequence label, anti-tag sequence label and combining of tag sequence label and concrete ASPE primer, can avoid cross reaction, realize the parallel detection in a plurality of SNP site.
2. the ASPE type specificity primer of the present invention's design has extraordinary specificity, can accurately distinguish the genotype of various types.
3. detection method step of the present invention is simple, two kinds of SNPs detect and can finish two amplifications that contain the target sequence in SNP site by a step multiplex PCR, many uncertain factors of existing in the complex operations processes such as repeated multiple times PCR have been avoided, thereby can improve the detection accuracy rate greatly, embodied qualitative, quantitative analysis feature of accurate while.
4. the needed time of detection method provided by the present invention is well below sequencing technologies commonly used, realistic especially application need.
5. not only to have overcome traditional solid phase chip susceptibility not high in the present invention, and the defective of the repeatability difference of detected result is improved existing liquid-phase chip technology simultaneously, makes prepared microballoon can be applicable to different test items, has very strong expansion.The fluorescent signal value that detects improves greatly, thereby makes the sensitivity that detects be further enhanced, and signal to noise ratio strengthens, and detected result more accurately and reliably.
Embodiment
Embodiment 1ADRB1 gene SNP detection liquid-phase chip mainly includes:
One, ASPE primer
Wild-type and mutant at the two kinds of common genotype A145G (rs1801252) and the C1165G (rs1801253) of ADRB1 gene design specific primer sequence respectively.The ASPE primer is made up of " Tag sequence+specific primer sequence ".The ASPE primer sequence is as shown in the table:
The ASPE primer sequence (Tag sequence+specific primer sequence) of table 1ADRB1 gene
Every the ASPE primer comprises two parts, and 5 ' end is the specificity tag sequence at anti-tag sequence on the corresponding microballoon, and 3 ' end is mutant or the special primer fragment (shown in above-mentioned table 1) of wild-type.All ASPE primers are synthetic by Shanghai Sangon Biological Engineering Technology And Service Co., Ltd.Every primer after synthetic is mixed with the stock solution of 100pmol/mL respectively with 10mmol/L Tris Buffer.
Two, the microballoon of anti-tag sequence bag quilt
According to designed ASPE Auele Specific Primer fragment, select the tag sequence, reduce between the anti-tag sequence of each microballoon to greatest extent and secondary structure that tag and ASPE Auele Specific Primer fragment may form, corresponding anti-tag sequence is as shown in table 2 on 4 kinds of microballoons numberings of selection and the microballoon:
Corresponding anti-tag sequence on table 2 microballoon numbering and the microballoon
Four kinds of microballoons selecting are available from U.S. Luminex company, with anti-tag sequence bag by on microballoon.Be connected with the spacerarm sequence of 5-10 T between anti-tag sequence and the microballoon, promptly add the spacerarm sequence of the preceding paragraph 5-10 T before each anti-tag sequence, the anti-tag sequence is synthetic by Shanghai Sangon Biological Engineering Technology And Service Co., Ltd.With synthetic anti-tag sequence sterilization ddH
2O is made into the stock solution of 100nmol/ml.Described spacerarm is to be used for anti-tag and microsphere surface is spaced apart or anti-tag is placed the sequence of hydrophilic environments.By the spacerarm sequence of suitable length is set between anti-tag sequence and microballoon, can reduce sterically hinderedly, improve the efficient of hybridization and the specificity of hybridization.Common spacerarm sequence comprises poly dT, i.e. poly (dT), and oligomerization four polyoxyethylene glycol and (CH2) n spacerarm (n 〉=3) are as (CH2) 12, (CH2) 18 etc.In addition, if exist poly (dA) to disturb, can also use poly (TTG) as spacerarm.Spacerarm of the present invention is preferably 5-10 T, and the process of microballoon bag quilt is as follows:
Get 5 * 10 respectively
6The carboxylated microballoon of individual above-mentioned numbering (available from Luminex company) is suspended in the MES solution of 50ul 0.1mol/L (pH4.5), adds 10ul synthetic anti-tag molecule (100nmol/ml).EDC (N-3-Dimethylaminopropyl-N-ethylcarbodiimide) (available from the Pierce Chemical company) working fluid of preparation 10ng/ml.Add the EDC working fluid of 2.5ul in the microballoon suspension, constant temperature was hatched 30 minutes, added the EDC working fluid of 2.5ul again, and constant temperature was hatched 30 minutes again.After reaction finished, the Tween-20 washing with 0.02% was once washed once with 0.1% SDS liquid again.The microballoon that is coated with the anti-tag sequence after the washing is resuspended in the Tris-EDTA solution [10mmol/L Tris (pH8.0)] of 100ul, and among the 1mmol/L EDTA, 2-8 ℃ keeps in Dark Place.
Three, amplify the primer of the target sequence that contains the mutational site
The primer that contains the mutational site target sequence at the SNP mutational site design amplification of two kinds of common genotype A145G of ADRB1 gene and C1165G.Wherein, the ask for number of A145G SNP sudden change in gene pool is rs1801252, the Serine that makes the 49th is replaced by glycine that (Ser → Gly), the ask for number of C1165G in gene pool be rs1801253, and the glycine that makes the 389th is by arginine replacement (Gly → Arg).Utilize the Primer5.0 design of amplification primers to (seeing Table 3), amplify two target sequences that contain the SNP site respectively.
Table 3 amplifies the primer of the target sequence with SNP site
All primers are synthetic by Shanghai Sangon Biological Engineering Technology And Service Co., Ltd.Every primer after synthetic is mixed with the stock solution of 100pmol/mL respectively with 10mmol/L TrisBuffer.
Embodiment 2 utilization embodiment 1 described ADRB1 gene test liquid-phase chip is to the detection of sample
The prescription of described various solution is as follows:
MES damping fluid (pH5.0) prescription (250ml) of 50mM:
| Reagent | The source | Final concentration | The consumption of every 250ml |
| MES(2[N-Morpholino] ethanesulfonic?acid) | Sigma?M-2933 | 0.05M | 2.44g |
| 5MNaOH | Fisher?SS256-500 | --- | 5 |
2 * Tm hybridization buffer
| Reagent | The source | Final concentration | The consumption of every 250ml |
| 1MTris-HCl，pH8.0 | SigmaT3038 | 0.2M | 50ml |
| 5MNaCl | Sigma?S5150 | 0.4M | 20ml |
| Triton?X-100 | Sigma?T8787 | 0.16％ | 0.4ml |
Be stored in 4 ℃ after the filtration.
The ExoSAP-IT test kit is available from U.S. USB company.
Biotin labeled dCTP is available from Shanghai Sangon Biological Engineering Technology And Service Co., Ltd.
One, the DNA extraction of sample:
With reference to " molecular cloning " methods involving, obtain DNA to be detected about DNA extraction.
Two, the pcr amplification of testing sample
Utilize two pairs of primers of Primer5.0 design, multiplex PCR one step amplifies two kinds of common genotype A145G containing the ADRB1 gene respectively and C1165G totally two target sequence, the product size is respectively 334bp and 234bp, and primer sequence (SEQ ID NO.13-16) is seen shown in the above-mentioned table 3.
At first prepare the multiple PCR primer working fluid: the primer stock solution 100ul that respectively gets SEQ ID NO.13-16 respectively mixes and is the multiple PCR primer working fluid in the 1.5ml Eppendorf tube.The multi-PRC reaction system is as follows:
2 * damping fluid (contains Mg
2+) 25ul
DNTP (each 2.5mmol/L) 4ul
Taq enzyme (5U/ul) 0.2ul
Multiple PCR primer working fluid (each 8.3pmol/mL) 6ul
Template DNA (10ng/ul) 2ul
ddH
2O 12.8ul
Be total to 50ul
The pcr amplification program is: 95 ℃ of 3min; 94 ℃ of 30s, 56 ℃ of 30s, 72 ℃ of 40s, 30 circulations; 72 ℃ of 10min; 4 ℃ of preservations are standby.
Three, the enzyme of PCR product is cut processing
1. get the reacted product of 7.5ul PCR, add 1ul 10 * SAP damping fluid, 1ul SAP enzyme and 0.5ul Exo-I enzyme;
2.37 ℃ hatch 15min, hatch 15min for 80 ℃, the enzyme that deactivation is unnecessary.The product that enzyme is cut after the processing is directly used in follow-up ASPE primer extension reaction.
Four, site-specific primer extension reaction (ASPE)
Utilize the ASPE primer of above-mentioned design to carry out primer extension reaction, in reaction process, mix biotin labeled dCTP, thereby make a plurality of biotin labeling on the reacted product band.
At first prepare blended ASPE primer working fluid: get the A145G of ADRB1 gene to be detected and C1165G corresponding wild type and mutant ASPE primer stock solution 10ul respectively in the 1.5ml Eppendorf tube, add 10mmol/L Tris Buffer and mend, mix and be ASPE mix primer working fluid to 200ul.The system of ASPE reaction is as follows:
10 * damping fluid 2ul
MgCl
2(50mmol/L) 0.5ul
Biotin-dCTP(400umol/L) 0.25ul
DATP, dGTP, dTTP mixed solution (each 100umol/L) 1ul
Tsp enzyme (5U/ul) 0.25ul
Blended ASPE primer working fluid (each 500nmol/L) 1ul
Enzyme is cut the pcr amplification product 5ul of processing
ddH
2O 10ul
Be total to 20ul
Response procedures is: 96 ℃ of 2min; 94 ℃ of 30s, 54 ℃ of 1min, 72 ℃ of 2min, 30 circulations; 4 ℃ of preservations are standby.
Five, hybridization
1. according to the ASPE primer of design, (microballoon concentration is 2.5 * 10 to the corresponding optimum microballoon of every group selection
5Individual/ml).
2. the microballoon of getting every kind of numbering of 1ul respectively is in the Eppendorf tube of 1.5ml;
3. microballoon is in 〉=centrifugal the 1-2min of 10000g;
4. supernatant discarded, microballoon is resuspended in 2 * Tm hybridization buffer of 100ul, the vortex mixing;
5. get the above-mentioned microballoon suspension of 25ul in the corresponding hole of 96 hole filter plates, control wells adds the ddH of 25ul
2O;
6. the ASPE reaction solution of getting 5-25ul is used ddH in corresponding hole
2O complements to 50ul;
7. encase 96 orifice plates with lucifuge with masking foil, 95 ℃ of 60s, 37 ℃ of 15min are hatched hybridization;
8. the microballoon after the hybridization is in 〉=centrifugal the 2-5min of 3000g;
9. remove supernatant, microballoon is resuspended in 1 * Tm hybridization buffer of 75ul;
10. microballoon is in 〉=centrifugal the 2-5min of 3000g;
11. microballoon is resuspended in 1 * Tm hybridization buffer of 75ul, adding 15ul concentration is Streptavidin-phycoerythrin (SA-PE) of 10ug/ml;
12.37 ℃ hatch 15min, on the Luminex instrument, detect.
Six, the result detects and data analysis
The reaction after product is by Luminex serial analysis instrument detecting.Detected result is shown in table 4, table 5 and table 6.
Fluorescent value (MFI) and data processing there are following requirement:
1. each site need have an allelotrope MFI at least greater than 300 and greater than 10 * PCR negative control MFI;
2.NET MFI=sample MFI-PCR negative control MFI (NET MFI less than 0 with 0 the expression);
3. satisfy the data of above two conditions, calculate sudden change ratio by following formula:
Sudden change ratio=mutant NET MFI ÷ (mutant NET MFI+ wild-type NET MFI)
4. rule of thumb the sudden change ratio of each detection site is determined threshold value (cut-off value), to divide wild-type homozygote, heterozygote and mutant homozygote.
Use present method to detect 20 increments ADRB1 gene SNP site originally, experimental data meets above-mentioned requirements, therefore can calculate their sudden change ratio.Being provided with of threshold value (cut-off value) is as follows: the sudden change ratio range is considered as the wild-type homozygote at 0%-20%; 30%-70% is considered as heterozygote; 80%-100% is considered as the anomaly homozygote.Compare with the liquid-phase chip result with the sequencing detection, calculate the identical rate of classifying method detected result provided by the present invention.Present method detects 20 increments ADRB1 genotype detection result and the sequencing result rate of coincideing originally and reaches 100%.As seen ADRB1 gene SNP detection liquid-phase chip provided by the present invention can detect the SNP type of ADRB1 exactly, and the result is reliable and stable.
Table 4 pattern detection result (MFI)
Table 5 sample ADRB1 transgenation ratio (%)
Table 6 sample ADRB1 gene mutation type analytical results
The liquid-phase chip of the ASPE primer that embodiment 3 is different is to the detection of ADRB1 gene SNP site
One, the design (selection of Tag sequence and Anti-Tag sequence) of liquid-phase chip preparation
Detecting liquid-phase chip with ADRB1 gene A 145G site mutation is example, respectively at the wild-type of A145G and the specific primer sequence of mutant design ASPE primer 3 ' end, the Tag sequence of ASPE primer 5 ' end then is selected from SEQ ID NO.1-SEQ IDNO.4, accordingly, bag is selected from SEQ ID NO.9-SEQ IDNO.12 by the anti-tag sequence with corresponding tag sequence complementary pairing on microballoon.Specific design is shown in following table (table 7).Synthetic, the anti-tag sequence bag of ASPE primer is described like embodiment 1 and embodiment 2 by microballoon, amplimer, detection method.
The design of table 7 liquid-phase chip preparation
Two, sample detection
Adopt the liquid-phase chip of above-mentioned design preparation, by embodiment 2 described testing processes and method sample 21-40 is detected, detected result is as follows:
Table 8 pattern detection result and Polymorphism Analysis
Other is at the liquid-phase chip in different SNP sites, and the ASPE primer uses different Tag sequences, and its result is still reliable and stable, and concrete data are omitted.
Embodiment 4 different interval arm liquid-phase chips detect the ADRB1 gene SNP
One, the design (selection of spacerarm) of liquid-phase chip preparation
Detection liquid-phase chip with ADRB1 gene C 1165G site mutation is an example, verifies the influence that different spacerarm liquid-phase chips detects the ADRB1 gene SNP.At the wild-type of C1165G and the specific primer sequence of mutant design ASPE primer 3 ' end, the Tag sequence of ASPE primer 5 ' end then is respectively SEQ ID NO.3 and SEQ ID NO.4, accordingly, bag is respectively SEQ ID NO.11 and SEQ ID NO.12 by the anti-tag sequence with corresponding tag sequence complementary pairing on microballoon.Verify the influence that different spacerarm liquid-phase chips detects the ADRB1 gene SNP, wherein, different spacerarms is (CH2) 12 or 5-10 T, and specific design is shown in following table (table 9).Synthetic, the anti-tag sequence bag of ASPE primer is described like embodiment 1 and embodiment 2 by microballoon, amplimer, detection method.
The design of table 9 liquid-phase chip preparation
Two, sample detection
Adopt the liquid-phase chip of above-mentioned design preparation, by embodiment 2 described testing processes and method sample 41-60 is detected, detected result is as follows:
Table 10 pattern detection result and Polymorphism Analysis
The spacerarm of embodiment 4 is the liquid-phase chip of (CH2) 12, its detected result is reliable and stable, and (detected result of other spacerarm is also like this, concrete data are omitted), and the hybridization rate of the spacerarm of preferred 5-10 T and hybridization specificity all are better than the liquid-phase chip of spacerarm for (CH2) 12, and the spacerarm of a 5-10 of the present invention T all is better than other spacerarms.
More than be at the specifying of possible embodiments of the present invention, but this embodiment is not in order to limiting claim of the present invention, does not allly break away from the equivalence that skill spirit of the present invention does and implement or change, all should be contained in the claim of the present invention.
Sequence table
＜110〉Guangzhou Yishan Biotechnology Co., Ltd.
＜120〉ADRB1 gene SNP detection specific primer and liquid-phase chip
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Claims (5)
1. an ADRB1 gene SNP detection liquid-phase chip is characterized in that, mainly includes:
(A) at the SNP site of ADRB1 gene, the ASPE primer of She Ji wild-type and mutant is right respectively: every kind of ASPE primer is made up of at the Auele Specific Primer in SNP site the tag sequence and the 3 ' end of 5 ' end, and described Auele Specific Primer is: at the SEQ ID NO.5 in A145G SNP site and SEQ ID NO.6 and/or at the SEQ ID NO.7 and the SEQ ID NO.8 in C1165G SNP site; Described tag sequence is selected from SEQ ID NO.1～SEQ ID NO.4;
(B) be coated with microballoon special anti-tag sequence, that have the different colours coding respectively, described anti-tag sequence is selected from the sequence among SEQID NO.9～SEQ ID NO.12, and described anti-tag sequence can be correspondingly with (A) in selected tag sequence complementary pairing, and described anti-tag sequence also is provided with the spacerarm sequence in the middle of being connected with microballoon;
(C) be used to amplify the amplimer of the ADRB1 gene target sequence in SNP site with A145G and/or C1165G.
2. ADRB1 gene SNP detection liquid-phase chip according to claim 1 is characterized in that,
Described amplimer is: at the SEQ ID NO.13 in A145G SNP site and SEQ ID NO.14 and/or at the SEQ ID NO.15 and the SEQ ID NO.16 in C1165G SNP site.
3. ADRB1 gene SNP detection liquid-phase chip according to claim 1, it is characterized in that described ASPE primer is: the sequence of forming at the sequence of forming by SEQ ID NO.1 and SEQ ID NO.5 in A145G SNP site and by SEQ ID NO.2 and SEQ IDNO.6 and/or at the sequence of forming by SEQ ID NO.3 and SEQ ID NO.7 in C1165G SNP site and the sequence of forming by SEQ ID NO.4 and SEQ ID NO.8.
4. according to each described ADRB1 gene SNP detection liquid-phase chip of claim 1-3, it is characterized in that described spacerarm sequence is 5-10 T.
5. be used for the Auele Specific Primer that the ADRB1 gene SNP detects, it is characterized in that, mainly include: at the SEQID NO.5 in A145G SNP site and SEQ ID NO.6 and/or at the SEQ ID NO.7 and the SEQ ID NO.8 in C1165G SNP site.
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| CN105441436A (en) * | 2015-10-10 | 2016-03-30 | 北京晋祺生物科技有限公司 | Individualized medication detection kit for receptor blocker |
| CN105586406A (en) * | 2016-01-15 | 2016-05-18 | 汪道文 | Method for detecting gene polymorphism of ADRB1 and GRK5 |
| CN106520979A (en) * | 2016-11-30 | 2017-03-22 | 武汉海吉力生物科技有限公司 | Nucleic acid, kit and method for detecting G1165C polymorphism of human ADRB1 gene |
| CN106834466A (en) * | 2017-02-08 | 2017-06-13 | 重庆迪威纳生物技术有限公司 | The method and kit in a kind of detection ADRB1 gene single nucleotide polymorphisms site |
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| CN101354343A (en) * | 2007-07-27 | 2009-01-28 | 上海主健生物工程有限公司 | Reagent kit for detecting adiposity genetic susceptibility |
| CN101560555A (en) * | 2009-03-18 | 2009-10-21 | 上海中优医药高科技有限公司 | Gene detection method used for individualistic education and health guidance of children |
| CN101565749A (en) * | 2009-04-15 | 2009-10-28 | 广州益善生物技术有限公司 | CYP2C19 and ABCB1 gene SNP detection liquid-phase chip and detection method thereof |
| CN101671739A (en) * | 2009-10-19 | 2010-03-17 | 广州益善生物技术有限公司 | Specific sequence, liquid phase chip and method for SNP detection of TPMT gene |
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| CN105441436A (en) * | 2015-10-10 | 2016-03-30 | 北京晋祺生物科技有限公司 | Individualized medication detection kit for receptor blocker |
| CN105441436B (en) * | 2015-10-10 | 2019-01-11 | 北京晋祺生物科技有限公司 | A kind of personalized medicine detection kit of beta blocker |
| CN105586406A (en) * | 2016-01-15 | 2016-05-18 | 汪道文 | Method for detecting gene polymorphism of ADRB1 and GRK5 |
| CN106520979A (en) * | 2016-11-30 | 2017-03-22 | 武汉海吉力生物科技有限公司 | Nucleic acid, kit and method for detecting G1165C polymorphism of human ADRB1 gene |
| CN106834466A (en) * | 2017-02-08 | 2017-06-13 | 重庆迪威纳生物技术有限公司 | The method and kit in a kind of detection ADRB1 gene single nucleotide polymorphisms site |
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| CN102021238B (en) | 2013-02-27 |
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