CN1847408A - Thrombocyte glucoprotein HPA-1 and HPA-2 genotype detecting chip and its use - Google Patents
Thrombocyte glucoprotein HPA-1 and HPA-2 genotype detecting chip and its use Download PDFInfo
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Abstract
The present invention provides one kind of gene chip for detecting thrombocyte glucoprotein HPA-1 and HPA-2 genotype, one kit including the gene chip and the method of detecting thrombocyte glucoprotein HPA-1 and HPA-2 genotype with the chip and the kit. The product and method of the present invention can detect the genotypes of the said genes so as to provide information essential for predicting disease risk related to the genes and provide measures essential for developing and evaluating the related medicines.
Description
(1) technical field
The present invention relates to the gene analysis test product, gene detecting chip specifically, saying so more specifically is applicable to and detects platelet glycoprotein HPA-1 and the genotypic gene detecting chip of HPA-2.The invention still further relates to platelet glycoprotein HPA-1 and the genotypic method of HPA-2 of detecting.
(2) background technology
Thrombocyte is seedless hemocyte.Hematoblastic function mainly is to promote hemostasis and quicken blood coagulation, and thrombocyte is safeguarded the function of capillary vessel wall integrity in addition simultaneously.Thrombocyte has the formation thrombus in hemostasis and coagulation process, stop up wound, discharges the functions such as the various factors relevant with blood coagulation.In quick mobile artery of blood and the thrombosed process of arteriole system, thrombocyte plays central role.
Platelet membrane contains multiple proteins, and these protein often are connected with a large amount of carbohydrate side chains and become glycoprotein.Platelet glycoprotein is not only most important to keeping thrombocyte form and integrity, and has constituted hematoblastic various acceptor, makes thrombocyte performance hemostasis and function associated.When the platelet glycoprotein gene morphs, can change platelet glycoprotein structure and expression level, so influence hematoblasticly stick, gathering and thrombosis.
Along with the progress of molecular biology and monoclonal antibody technique, the gene order of many platelet glycoproteins is confirmed, and makes the further investigation to the platelet glycoprotein polymorphism become possibility.After this, aspect biological chemistry essence, function and the molecular biology research thereof of platelet glycoprotein, making great progress.(human platelet alloantigen, HPA), each antigen systems is to be controlled by the codominance diallele human thrombocyte allogenic antigen system on the platelet glycoprotein gene.Gene order to platelet glycoprotein IIIa and glycoprotein ibalpha is discovered, have HPA-1 1a/1b allelotrope on No. 2 exon of platelet glycoprotein IIIa gene, there is HPA-2 2a/2b allelotrope in the receptor binding domain of platelet glycoprotein Ib gene.
The thrombocyte infusion has important effect in clinical.Yet, owing to have specificity platelet antigen on the thrombocyte, often do not take place because of antigen conforms to that the thrombocyte infusion is invalid during clinical use, post-transfusion purpura etc.Even more serious is that many patients are owing to thrombopenia, and infusion is invalid, has influenced carrying out and carrying out of clinical treatment, even has taken place hemorrhage, dead.Therefore, detected the specificity platelet antigen of acceptor and donor in the past, and helped avoid because of antigen and do not conform to the adverse consequences that causes at platelet transfusion.In addition, have bibliographical information platelet glycoprotein HPA-1, HPA-2 polymorphism relevant with myocardial infarction and ischemic cerebrovascular disease in recent years, HPA-1, HPA-2 genovariation are myocardial infarction and risk factor for isehemic stroke.Thereby it is significant to the exploitation of clinical individualized treatment, medical research and novel antithrombotic reagent-platelet glycoprotein inhibitor, evaluation etc. to detect HPA-1, HPA-2 genovariation quickly and accurately.
In recent years, the research of detection HPA-1, HPA-2 genotype method receives much concern.But present detection method mainly adopts polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), craft or methods such as automatic sequencing, sequence specific primers PCR, complex operation not only, sense cycle is long, and the factor that influences detected result is many, wayward, be difficult to satisfy requirement of actual application, make drug research unit also can't extensively carry out relevant HPA-1 and the genotypic detection of HPA-2 so far with clinical.
Gene chip is one of great science and technology progress of the tool characteristics of the times that occurred in high-tech area in recent years.It is the gene probe (oligonucleotide probe, cDNA clone, PCR product etc.) with a large amount of gene informations in the energy reflected sample, being fixed on solid support (as the slide glass of aldehyde radical, amino, sulfydryl, carboxyl isoreactivity base group modification or silicon chip, nylon membrane, nitrocellulose filter) in order goes up and forms array, by carrying out hybridization with actual sample (or amplified production), only need once experiment, but obtain the information of all genes to be checked with regard to high-throughput.The characteristics of this parallel detection, high information flux make its application at aspects such as genetic expression, gene pleiomorphism detections be subjected to extensive attention.Detect HPA-1 and HPA-2 genotype with gene chip, performance gene chip detecting operation is simple, characteristic of accurate as a result, has important practical significance for drug research unit and clinical related application.
(3) summary of the invention
The present invention is directed to the deficiencies in the prior art, a kind of genotypic gene detecting chip of platelet glycoprotein HPA-1, HPA-2 that is used to detect is provided.
The present invention also provides a kind of and detects platelet glycoprotein HPA-1, the genotypic detection method of HPA-2 with gene detecting chip.
Gene chip provided by the invention comprises solid support and is fixed on oligonucleotide probe on the described solid support in order, and described oligonucleotide probe comprises one section amido modified 16 poly-poly-deoxythymidylic acids (poly-dT) at 5 ' end, and each probe sequence is:
HPA1-1a allele：NH
2-5’-tttttttttttttttt-ggtgagcccGgaggcagggcct-3’
(SEQ ID NO：1)
HPA1-1b allele：NH
2-5’-tttttttttttttttt-ggtgagcccAgaggcagggcct-3’
(SEQ ID NO：2)
HPA2-2a allele：NH
2-5’-tttttttttttttttt-gggctcctgaTgcccacacccaa-3’
(SEQ ID NO：3)
HPA2-2b allele：NH
2-5’-tttttttttttttttt-gggctcctgaCgcccacacccaa-3’
(SEQ ID NO：4)
In order to strengthen the intensity of detection signal, improve the accuracy rate of detected result, the base that capitalization is represented in described HPA-1 and the HPA-2 mutational site sequence preferably is positioned at the middle part of described probe.
Sequence oligonucleotide probe of the present invention is to design acquisition according to the continuous nucleotide sequence that comprises HPA-1 and HPA-2 polymorphic site in following sequence or its complementary sequence:
HPA1-1a allele：ctccttcaggtcacagcgaggtgagcccGgaggcagggcctgtaagacagga
(SEQ ID NO：5)
HPA1-1b allele：ctccttcaggtcacagcgaggtgagcccAgaggcagggcetgtaagacagga
(SEQ ID NO：6)
HPA2-2a allele：gaagaccctgcccccagggctcctgaTgcccacacccaagctggagaagctc
(SEQ ID NO：7)
HPA2-2b allele：gaagaccctgcccccagggctcctgaCgcccacacccaagctggagaagctc
(SEQ ID NO：8)
Described solid support can adopt the common used material of range gene chip field, as but be not limited to nylon membrane, the slide of slide of modifying through active group or silicon chip, unmodified, plastic sheet etc.
The slide of above-mentioned modification or the active group of silicon chip as but be not limited to aldehyde radical, amino, isothiocyanic acid base etc., preferred aldehyde group modified slide or silicon chip.
The preparation of gene chip of the present invention can be carried out according to the conventional manufacture method of biochip.For example, if what solid support adopted is to modify slide or silicon chip, 5 of probe ' end contains amido modified poly-dT string, oligonucleotide probe can be mixed with solution, with point sample instrument its point is being modified slide or silicon chip then, be arranged in predetermined sequence or array, spending the night by placement then fixes, and just can obtain gene chip of the present invention.If it is amido modified that oligonucleotide probe does not contain, then its preparation method also can reference: Wang Shenwu chief editor's " gene diagnosis technology-on-radiation operational manual " and the upright people of horse, Jiang Zhonghua edits. biochip. Beijing: Chemical Industry Press, 2000,1-130.
It is a kind of non-to be diagnosed as purpose detection HPA-1 and the genotypic method of HPA-2 that the present invention also provides, and may further comprise the steps:
(1) preparation chromosomal DNA;
(2) with the gene fragment that comprises HPA-1 and HPA-2 polymorphic site in polymerase chain reaction (PCR) method amplification platelet glycoprotein IIIa and the glycoprotein ibalpha gene, comprise design of primers;
(3) the above-mentioned amplified production of mark;
(4) with the amplified production of above-mentioned mark and above-mentioned gene chip hybridization;
(5) hybridization signal of detection gene chip.
The method that preparation is used for the chromosomal DNA of pcr amplification has a lot, can prepare from whole blood, also can prepare from root of hair, can also prepare from the cast of oral mucosa.Concrete grammar can be consulted the fourth promise of shaking, the Su Mingquan chief editor. " clinical PCR gene diagnosis technology ", the .1998 of world book publishing company first version.
With the specific segmental method of PCR method amplification chromogene has been techniques well known.Key wherein is design of primers, and method, the software of relevant design of primers all can obtain from commercial channels.The amplimer of HPA-1 that the present invention relates to and HPA-2 gene pleiomorphism can independently be finished by the user according to above method and relevant literature method with system.As in Mu KB this, in F. takes, R. gibbs etc. chief editor " PCR polymerase chain reaction ", Science Press.
Amplified production is carried out the method that mark can increase by the primer that adopts 5 ' end tape label group, the method of mononucleotide that also can be by mixing the tape label group in amplification procedure is realized that described labelling groups includes but not limited to: digoxin molecule (DIG), biotin molecule (Bio), fluorescein and derivative molecular (FITC etc.) thereof, other fluorescence molecules (as Cy3, Cy5 etc.), alkaline phosphatase (AP), horseradish peroxidase (HRP) etc.The detection method of these marks and marking method thereof and each marker all has been routine techniques well-known in the art, also can be with reference to Wang Shenwu chief editor's " gene diagnosis technology-on-radiation operational manual "; J. Sa nurse Brooker, E.F. is the Ritchie not, T. Manny A Disi chief editor, " molecular cloning experiment guide ", Science Press, nineteen ninety-five; Horse stands the people, Jiang Zhonghua chief editor, " biochip ", Beijing: Chemical Industry Press, 2000,1-130.
The sex change of amplified production can be adopted conventional denaturation method, and the present invention preferably is heated to 94～98 ℃, and is incubated 2～10min, puts on ice rapidly then.
The amplified production of getting an amount of sex change adds in the hybridization buffer and gene chip hybridization.During with gene chip hybridization, can earlier gene chip and prehybridization damping fluid be carried out prehybridization.
Solid-phase hybridization between amplified production of the present invention and the gene chip carries out according to the classical way of this area, and the general personnel in this area determine relevant damping fluid, probe and the optimum condition of sample concentration, prehybridization temperature, hybridization temperature and time etc. easily according to experience.Perhaps also can be with reference to Wang Shenwu chief editor's " gene diagnosis technology-on-radiation operational manual "; J. Sa nurse Brooker, E.F. is the Ritchie not, T. Manny A Disi chief editor, " molecular cloning experiment guide ", Science Press, nineteen ninety-five.
Obtain according to information such as the position of marking signal on gene chip, intensity then and treat measurement information.The method of detection gene chip hybridization signal of the present invention is based on alkaline phosphatase or the tetrazole indigo plant (NBT) of horseradish peroxidase enzyme catalytic and the color reaction of 5-bromo-4-chloro-3-indolol-phosphoric acid-4-toluene amine salt (BCIP) or tetramethyl benzidine (TMB) that is combined with each other with anti-biotin antibodies or affinity element or streptavidin or anti digoxin antibody or anti-fluorescein antibody.Concrete grammar can be with reference to Wang Shenwu chief editor's " gene diagnosis technology-on-radiation operational manual ".If amplified production fluorophor mark, also can be directly obtain and treat measurement information with fluorescence detection device (as laser confocal scanning instrument Scanarray3000 etc.).
The present invention also provides a kind of be used to detect HPA-1 and the genotypic test kit of HPA-2, this test kit comprises said gene chip of the present invention and working instructions, and working instructions comprise non-to be diagnosed as the associated viscera that purpose detects HPA-1 and genotypic method of HPA-2 and above-mentioned prior art.
Gene chip provided by the invention and test kit thereof can be used for HPA-1 and HPA-2 genotype, and this provides a kind of simple and easy to do solution for related application such as drug research unit and clinical treatments targetedly.
(4) description of drawings
Fig. 1: a kind of point sample array on the gene chip of the present invention;
Fig. 2: the photo that detects HPA-1 and HPA-2 genotype gained result with gene chip of the present invention.
(5) embodiment
Following examples are to more detailed description of the present invention, rather than limiting the scope of the invention.
Embodiment 1: the preparation of gene chip
Purchase aldehyde group modified slide glass (production code member: BSM03011, Baiao Science and Technology Co. Ltd., Shanghai).The following probe of synthetic (Shanghai Sangon Biological Engineering Technology And Service Co., Ltd), water is dissolved into (100pmol/ul) concentration, uses 2 * point sample buffer (production code member: BST02010, Baiao Science and Technology Co. Ltd., Shanghai) geometric ratio to mix then.Then, with the array of the described method point system of the GMS417 point sample instrument by specification of Affymetrix company as Fig. 1.Room temperature is placed and is spent the night then.
Wherein each probe sequence is:
HPA1-1a allele：NH
2-5’-tttttttttttttttt-ggtgagcccGgaggcagggcct-3’
(SEQ ID NO：1)
HPA1-1b allele：NH
2-5’-tttttttttttttttt-ggtgagcccAgaggcagggcct-3’
(SEQ ID NO：2)
HPA2-2a allele：NH
2-5’-tttttttttttttttt-gggctcctgaTgcccacacccaa-3’
(SEQ ID NO：3)
HPA2-2b allele：NH
2-5’-tttttttttttttttt-gggctcctgaCgcccacacccaa-3’
(SEQ ID NO：4)
Embodiment 2: the preparation of chromosomal DNA
Get person's whole blood 500 μ l to be checked, extract with sterile hypodermic needle for single use, be collected in the aseptic 1.5ml centrifuge tube of the 2%EDTA solution that contains 50ul, will manage and cover, the several that turns upside down makes mixing.Get aseptic 1.5ml centrifuge tube, to whole blood to be checked 100 μ l that wherein add mixing and pure water 300 μ l, abundant mixing, room temperature left standstill 8 minutes; Centrifuge tube is put in the whizzer centrifugal 1 minute of 6000g; Take out centrifuge tube, the supernatant liquor that inclines, the visible a small amount of white precipitate in centrifuge tube bottom; Add extract (production code member: BST01010, Baiao Science and Technology Co. Ltd., Shanghai) 60 μ l in centrifuge tube, fully vibration makes resolution of precipitate; In the boiling water bath 15 minutes; Take out centrifuge tube, put in the whizzer, 12, centrifugal 15 minutes of 000g.Centrifugal back supernatant liquor can be directly used in pcr amplification.
Embodiment 3: with PCR method amplification HPA-1 and HPA-2 gene pleiomorphism fragment
Entrust the synthetic following primer of Shanghai Sangon Biological Engineering Technology And Service Co., Ltd:
HPA1 upstream primer: 5 '-ctgacttgagtgacctgggag-3 '
(SEQ ID NO：9)
HPA1 downstream primer: Biotin-5 '-cttagctattgggaagtggtagg-3 '
(SEQ ID NO：10)
HPA2 upstream primer: 5 '-gacgtctccttcaaccggctg-3 '
(SEQ ID NO：11)
HPA2 downstream primer: 5 '-Biotin-ggtattgtatacagcgagttctcttg-3 '
(SEQ ID NO：12)
Then with water dissolution and be diluted to 10pmol/ μ l.With the Taq enzyme (production code member of purchasing: Lot, CE1101-1, TaKaRa), 10 * damping fluid (production code member: Lot, A2401-2, TaKaRa), the pcr amplification template of dNTP (production code member: D0056, Shanghai Sangon Biological Engineering Technology And Service Co., Ltd), pure water and embodiment 2 acquisitions is pressed following formulated pcr amplification system:
| Reaction system | Volume (μ l) |
| 10 * damping fluid | 2.5 |
| DNTP (each 2.5mM) | 2.5 |
| Primer 1 (10pmol/ul) | 1 |
| Primer 2 (10pmol/ul) | 1 |
| Taq enzyme (2.5U/ul) | 0.8 |
| H 2O | 14.2 |
| Template | 3 |
| Cumulative volume | 25μl |
Increase by following program with pcr amplification instrument Tc-25/H (Dahe Thermomagnetic Electronic Co., Ltd., Hangzhou): 94 ℃, 5min, then by 94 ℃ of 25sec, 56 ℃ of 25sec, 72 ℃ of 25sec do 40 circulations, last 72 ℃ of 5min.
The hybridization of embodiment 4:PCR product and gene chip
Adopt the chip hybridization test kit and the chip colouring reagents box of Baiao Science and Technology Co. Ltd., Shanghai to carry out this cross experiment.Described chip hybridization test kit (production code member: BST05010, Baiao Science and Technology Co. Ltd., Shanghai) comprises: prehybridization solution, hybridization buffer, washing lotion 1, reaction cabin.Described chip colouring reagents box (production code member: BST06010, Baiao Science and Technology Co. Ltd., Shanghai) comprises: antibody liquid, washing lotion 2, washing lotion 3, colour developing liquid.
(1) with the gene chip of embodiment 1 prehybridization solution prehybridization 5min, removes prehybridization solution then.The prehybridization temperature of chip is 39 ℃.
(2) pcr amplification product that embodiment 3 is obtained mixes back 98 ℃ of sex change 5min earlier, gets 10 μ l then and adds to mixing in the 190 μ l hybridization buffers, and the gene chip that finishes with prehybridization carries out 30 minutes hybridizations, and hybridization temperature is 39 ℃.
(3) gene chip that hybridization is finished washes twice, each 10 minutes with the washing lotion 1 that is preheated to hybridization temperature;
(4) washed 2 minutes with washing lotion 2 room temperatures then;
(5) with gene chip and antibody liquid room temperature reaction 20 minutes;
(6) then gene chip was washed 5 minutes with washing lotion 2 room temperatures, repeated this step more once; Washed 2 minutes with washing lotion 3 room temperatures again.
(7) on gene chip, add colour developing liquid 200 μ l, placed 40 minutes for 39 ℃.Water is rinsed well then, dries.
Embodiment 5: the detection of gene chip hybridization signal
Place Baio BE3.0 biochip to distinguish the detected result that obtains after instrument (BSE01021, Baiao Science and Technology Co. Ltd., Shanghai) upward scans as shown in Figure 2 the gene chip of hybridizing and washing finishes.The result shows that person under inspection's HPA1 and HPA2 genotype are: HPA1:1a/1b; HPA2:2a.Detected result meets fully through the sequencing result checking.
Sequence table
＜110〉Zhang Yong
＜120〉a kind of platelet glycoprotein HPA-1 and HPA-2 genotype detecting chip and uses thereof
<170>Patent In3.1
<212>PRT
<400>1-12
HPA1-1a allele：NH
2-5’-tttttttttttttttt-ggtgagcccGgaggcagggcct-3’
(SEQ ID NO：1)
HPA1-1b allele：NH
2-5’-tttttttttttttttt-ggtgagcccAgaggcagggcct-3’
(SEQ ID NO：2)
HPA2-2a allele：NH
2-5’-tttttttttttttttt-gggctcctgaTgcccacacccaa-3’
(SEQ ID NO：3)
HPA2-2b allele：NH
2-5’-tttttttttttttttt-gggctcctgaCgcccacacccaa-3’
(SEQ ID NO：4)
HPA1-1a allele：ctccttcaggtcacagcgaggtgagcccGgaggcagggcctgtaagacagga
(SEQ ID NO：5)
HPA1-1b allele：ctccttcaggtcacagcgaggtgagcccAgaggcagggcctgtaagacagga
(SEQ ID NO：6)
HPA2-2a allele：gaagaccctgcccccagggctcctgaTgcccacacccaagctggagaagctc
(SEQ ID NO：7)
HPA2-2b allele：gaagaccctgcccccagggctcctgaCgcccacacccaagctggagaagctc
(SEQ ID NO：8)
HPA1 upstream primer: 5 '-ctgacttgagtgacctgggag-3 '
(SEQ ID NO：9)
HPA1 downstream primer: Biotin-5 '-cttagctattgggaagtggtagg-3 '
(SEQ ID NO：10)
HPA2 upstream primer: 5 '-gacgtctccttcaaccggctg-3 '
(SEQ ID NO：11)
HPA2 downstream primer: 5 '-Biotin-ggtattgtatacagcgagttctcttg-3 '
(SEQ ID NO：12)
Claims (7)
1, a kind of platelet glycoprotein HPA-1 and HPA-2 genotype detecting chip, comprise solid support and be fixed on oligonucleotide probe on the described solid support in order, described oligonucleotide probe comprises one section 16 amido modified poly-deoxythymidylic acid that gather at 5 ' end, and each probe sequence is:
HPA1-1a allele：NH
2-5’-tttttttttttttttt-ggtgagcccGgaggcagggcct-3’
(SEQ ID NO：1)
HPA1-1b allele：NH
2-5’-tttttttttttttttt-ggtgagcccAgaggcagggcct-3’
(SEQ ID NO：2)
HPA2-2a allele：NH
2-5’-tttttttttttttttt-gggctcctgaTgcccacacccaa-3’
(SEQ ID NO：3)
HPA2-2b allele：NH
2-5’-tttttttttttttttt-gggctcctgaCgcccacacccaa-3’
(SEQ ID NO：4)。
2, platelet glycoprotein HPA-1 as claimed in claim 1 and HPA-2 genotype detecting chip is characterized in that described solid support adopts nylon membrane, the slide or the plastic sheet of the slide modified through active group or silicon chip, unmodified.
3, platelet glycoprotein HPA-1 as claimed in claim 2 and HPA-2 genotype detecting chip is characterized in that solid support is aldehyde group modified slide or silicon chip.
4, platelet glycoprotein HPA-1 as claimed in claim 2 and HPA-2 genotype detecting chip is characterized in that the slide modified or the active group of silicon chip are aldehyde radical, amino, isothiocyanic acid base.
5, a kind of non-to be diagnosed as purpose detection HPA-1 and the genotypic method of HPA-2, may further comprise the steps:
(1) preparation chromosomal DNA;
(2) with the gene fragment that comprises HPA-1 and HPA-2 polymorphic site in polymerase chain reaction method amplification platelet glycoprotein IIIa and the glycoprotein ibalpha gene, comprise design of primers;
(3) the above-mentioned amplified production of mark;
(4) with the amplified production of above-mentioned mark and the gene chip hybridization of claim 1;
(5) hybridization signal of detection gene chip.
6, as claimed in claim 5 non-to be diagnosed as purpose detection HPA-1 and the genotypic method of HPA-2, it is characterized in that the primer sequence described in the step (2) is as follows:
HPA1 upstream primer: 5 '-ctgacttgagtgacctgggag-3 '
SEQ ID NO：9
HPA1 downstream primer: Biotin-5 '-cttagctattgggaagtggtagg-3 '
SEQ ID NO：10
HPA2 upstream primer: 5 '-gacgtctccttcaaccggctg-3 '
SEQ ID NO：11
HPA2 downstream primer: 5 '-Biotin-ggtattgtatacagcgagttctcttg-3 '
SEQ ID NO：12。
7, a kind ofly be used to detect HPA-1 and the genotypic test kit of HPA-2, comprise the gene chip and the working instructions of claim 1, working instructions comprise the content of the detection method of claim 6 and relevant therewith prior art.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2006100423995A CN100371460C (en) | 2006-02-20 | 2006-02-20 | Thrombocyte glucoprotein HPA-1 and HPA-2 genotype detecting chip and its use |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2006100423995A CN100371460C (en) | 2006-02-20 | 2006-02-20 | Thrombocyte glucoprotein HPA-1 and HPA-2 genotype detecting chip and its use |
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|---|---|
| CN1847408A true CN1847408A (en) | 2006-10-18 |
| CN100371460C CN100371460C (en) | 2008-02-27 |
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|---|---|---|---|
| CNB2006100423995A Expired - Fee Related CN100371460C (en) | 2006-02-20 | 2006-02-20 | Thrombocyte glucoprotein HPA-1 and HPA-2 genotype detecting chip and its use |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101845520A (en) * | 2010-05-11 | 2010-09-29 | 上海市血液中心 | HPA allelic gene typing detection reagent kit |
| CN102230017A (en) * | 2011-06-23 | 2011-11-02 | 中国人民解放军第四军医大学 | Human platelet antigen genotyping liquid chip and human platelet antigen genotyping detection method thereof |
| CN111235261A (en) * | 2019-12-20 | 2020-06-05 | 江苏伟禾生物科技有限公司 | Kit for detecting human platelet-specific antigen HPA 1-29 genotyping |
| CN113249493A (en) * | 2021-03-05 | 2021-08-13 | 浙江省血液中心 | Real-time fluorescence PCR method, probe, primer and kit for typing human platelet alloantigen system alleles |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1591534A1 (en) * | 2004-04-01 | 2005-11-02 | Stichting Sanquin Bloedvoorziening | A method of genotyping blood cell antigens and a kit suitable for genotyping blood cell antigens |
-
2006
- 2006-02-20 CN CNB2006100423995A patent/CN100371460C/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101845520A (en) * | 2010-05-11 | 2010-09-29 | 上海市血液中心 | HPA allelic gene typing detection reagent kit |
| CN101845520B (en) * | 2010-05-11 | 2012-06-06 | 上海市血液中心 | HPA allelic gene typing detection reagent kit |
| CN102230017A (en) * | 2011-06-23 | 2011-11-02 | 中国人民解放军第四军医大学 | Human platelet antigen genotyping liquid chip and human platelet antigen genotyping detection method thereof |
| CN111235261A (en) * | 2019-12-20 | 2020-06-05 | 江苏伟禾生物科技有限公司 | Kit for detecting human platelet-specific antigen HPA 1-29 genotyping |
| CN111235261B (en) * | 2019-12-20 | 2022-11-04 | 江苏伟禾生物科技有限公司 | Kit for detecting human platelet-specific antigen HPA 1-29 genotyping |
| CN113249493A (en) * | 2021-03-05 | 2021-08-13 | 浙江省血液中心 | Real-time fluorescence PCR method, probe, primer and kit for typing human platelet alloantigen system alleles |
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| Publication number | Publication date |
|---|---|
| CN100371460C (en) | 2008-02-27 |
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