CN113981068B - 一种同时检测心脑血管药物代谢能力基因的引物探针系统及检测试剂盒 - Google Patents
一种同时检测心脑血管药物代谢能力基因的引物探针系统及检测试剂盒 Download PDFInfo
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Abstract
本发明公开了一种同时检测心脑血管药物代谢能力基因的引物探针系统及检测试剂盒，其中，所述引物探针系统同时检测以下24个SNP位点：rs5065、rs12041331、rs5186、rs730012、rs776746、rs2070744、rs12248560、rs4986893、rs4244285、rs1057910、rs1801253、rs2238032、rs2239050、rs2239128、rs2306283、rs4149056、rs671、rs1799752、rs16960228、rs4149601、rs429358、rs7412、rs1065852和rs9923231的基因多态性。本发明采用全新的次黄嘌呤替代技术优化了探针结构，同时加强了探针修饰，因此整体的引物探针系统的特异性更高，检测准确度更好，本发明的探针系统和检测试剂盒具有通量高、准确、特异性高的特点，将多种目标基因联合检测，单位时间和反应空内同时获得多个代谢基因的检测结果。
Description
技术领域
本发明涉及一种同时检测心脑血管药物代谢能力基因的引物探针系统及检测试剂盒，属于基因检测领域。
背景技术
目前心血管疾病已成为人类健康的第一大杀手，市面上针对心脑血管疾病的药品种类很多，根据药理机制不同，可大致如下分类：1、抗血小板聚集药物，如阿司匹林、氯吡格雷、奥扎格雷。但是脑出血时，这类药物禁用。2、调脂稳定斑块的他汀类药物，阿托伐他汀、瑞舒伐他汀等。3、改善细胞代谢的药物，如曲美他嗪、辅酶Q10等。4、降低氧耗和氧自由基的药物，如美托洛尔、比索洛尔、依达拉奉等。上述药物的选择与联合应用，在基因水平参数等指导下“量体裁衣”。基因芯片技术在心血管疾病中的研究可以同时快速地分析大量的基因信息，方便心血管疾病的检测，在心血管疾病诊断和用药等方面都具有重要的价值。
目前，对于基因多态性检测的方法主要有直接测序法、高分辨率熔解曲线法、等位基因特异性扩增法、taqman荧光探针法等。其中，此类方法普遍扩增通量不高。因此，需要建立一种简单、价格低廉的高通量检测基因多态性的方法。基因芯片(Gene Chip)通常指DNA芯片，其基本原理是将大量寡核苷酸分子固定于支持物上，然后与标记的样品进行杂交，通过检测杂交信号的强弱进而判断样品中靶分子的数量，具有高通量、简便、微缩、集约化、平行化等优点。基因芯片的概念现已泛化到生物芯片、微阵列(Microarray)、DNA芯片(DNAchip)，甚至蛋白芯片。心血管病是一种与多基因有关的复杂疾病，其发生、发展与多基因位点变异有关。基因芯片技术在心血管疾病可进行多位点、高通量的药物相关的基因多态性检测。
研究显示，多数基因多态性位点与心血管疾病药物代谢均有一定的关联性，但单个位点与药物代谢之间的强关联性不足，需要多个位点同时检测后，汇总测序结果进行综合判断，因此，生物芯片将成为未来的心血管疾病预测、辅助诊断、药物筛选等方面的重要参考依据。将研究成果产业化，是目前摆在基因芯片领域的一个主要问题。但是目前的基因芯片与其他检测方法相比存在检测灵敏度较低的问题，多个基因同时互补匹配有可能会造成假阳性或假阴性的产生。因此为了将生物芯片在心血管疾病领域产业化，需要结合现有的基因芯片技术和已知基因多态性位点进行筛选研究，并针对性的改善灵敏度低的问题，设计开发出一款可以产业化应用的心脑血管药物代谢相关的基因多态性生物芯片是十分必要的。
发明内容
针对现有技术存在的上述问题，本发明的目的是获得一种同时检测心脑血管药物代谢能力基因的引物探针系统及检测试剂盒。
为实现上述发明目的，本发明采用的同时检测心脑血管药物代谢能力基因的引物探针系统及检测试剂盒的技术方案如下：
所述引物探针系统可同时检测以下24个SNP位点：rs5065、rs12041331、rs5186、rs730012、rs776746、rs2070744、rs12248560、rs4986893、rs4244285、rs1057910、rs1801253、rs2238032、rs2239050、rs2239128、rs2306283、rs4149056、rs671、rs1799752、rs16960228、rs4149601、rs429358、rs7412、rs1065852、rs9923231的基因多态性。
所述检测试剂盒包括扩增反应液、检测反应液及阳性对照，所述扩增反应液包括如序列表SEQ ID NO:1～SEQ ID NO:23所示的上游引物序列和如序列表SEQ ID NO:24～SEQ ID NO:46所示的下游引物序列，所述下游引物序列的5`端均采用CY5标记。定位探针序列如序列表SEQ ID NO:47所示。具体的序列表如下：
所述的24个SNP位点rs5065、rs12041331、rs5186、rs730012、rs776746、rs2070744、rs12248560、rs4986893、rs4244285、rs1057910、rs1801253、rs2238032、rs2239050、rs2239128、rs2306283、rs4149056、rs671、rs1799752、rs16960228、rs4149601、rs429358、rs7412、rs1065852、rs9923231的检测探针序列如序列表SEQ ID NO:48～SEQ IDNO:95所示；所述阳性探针序列如序列表SEQ ID NO:96～SEQ ID NO:119所示。所述定位参照序列如序列表SEQ ID NO:120所示。
更优选的，所述的检测探针多态性分辨位点采用LNA标记，“+”表示，可提高单碱基的分辨率。为进一步提高检测探针的分辨率探针中有碱基采用次黄嘌呤替代(I)，替代后的探针对特异性更优，同时可以大大提高对捕获效率和减少探针的二级结构的形成。
更优选的，所述的检测探针和阳性探针的5’端采用C12氨基修饰和(dT)15修饰，有效减少了探针与PCR产物在特异性结合时的空间位阻效应。
优选的，所述的扩增反应液，包括特异性扩增引物，还包括多重PCR预混液(MultiEnzyme Mix)，Multi Buffer Mix、Multi酶系Mix、Solution II为Multi Enzyme Mix组分；
优选的，所述的阳性对照，包括24个位点浓度为10E5copies/ul的突变型和野生型质粒等比混合物，对未知样本的型别判定提供参考，同时对反应液的有效性进行质控。
优选的，所述的反应体积为50ul，反应条件为：预变性95℃，10min；变性95℃，15s，退火60℃，20s，延伸72℃，30s，扩增35个循环；最终延伸72℃，7min；保存4℃∞。
本发明还公开了一种采用上述检测试剂盒的心脑血管药物相关检测相关的基因多态性检测方法，所述检测方法包括以下步骤：
a.将所述扩增反应液与4ul待测DNA均匀进行PCR扩增；
b.PCR产物及杂交液等比例混匀后在95℃条件下变性10min后，立即置于冰浴中5min；
c.取变性的扩增产物10μl，加入检测试剂盒反应区中使其分布均匀，将检测试剂盒平置于杂交盒中，在待定杂交温度的水浴中杂交1小时；
d.将检测试剂盒在SSC洗涤液中漂洗；
e.数据处理和图像分析。
本发明还公开了一种采用上述的同时检测心脑血管药物代谢能力基因的检测试剂盒，其中，所述检测试剂盒包括扩增反应液、检测反应液及阳性对照，所述检测反应液中包括上述的引物探针系统。
与现有技术相比，本发明采用全新的次黄嘌呤替代技术优化了探针结构，同时加强了探针修饰，因此整体的引物探针系统的特异性更高，检测准确度更好。并且，本发明的探针系统和检测试剂盒具有通量高、准确、特异性高的特点，将多种目标基因联合检测，单位时间和反应空内同时获得多个代谢基因的检测结果。本发明的检测方法快速、准确及超高灵敏度地检测24个SNP位点，根据检测结果，可实时、快速监测心脑血管药物代谢的基因位点的基因型，及时评估待测者的状况，为临床医师提供辅助治疗建议。
附图说明
图1是本发明提供的基因芯片生产工艺流程图；
图2是本发明提供的基因芯片矩阵示意图；
图3是本发明提供的阳性对照芯片检测结果图。
具体实施方式
下面结合实施例对本发明提供的同时检测心脑血管药物代谢能力基因的引物探针系统及检测试剂盒作进一步详细、完整地说明。下面描述的实施例是示例性的，仅用于解释本发明，而不能理解为对本发明的限制。
下述实施例中的实验方法，如无特殊说明，均为常规方法。下述实施例中所用的实验材料如无特殊说明，均为市场购买得到。
实施例1、检测试剂盒的制备
本发明的检测试剂盒针对24个SNP位点rs5065、rs12041331、rs5186、rs730012、rs776746、rs2070744、rs12248560、rs4986893、rs4244285、rs1057910、rs1801253、rs2238032、rs2239050、rs2239128、rs2306283、rs4149056、rs671、rs1799752、rs16960228、rs4149601、rs429358、rs7412、rs1065852、rs9923231的多态性设计特异性扩增引物。基于多重扩增技术设计引物和芯片探针是本发明的关键之一，基因多态性序列以Genebank内的公开序列为准。
(一)本实施例的扩增引物序列如下：
所述检测试剂盒包括扩增反应液、检测反应液及阳性对照，所述扩增反应液包括如序列表SEQ ID NO:1～SEQ ID NO:23所示的上游引物序列和如序列表SEQ ID NO:24～SEQ ID NO:46所示的下游引物序列，所述下游引物序列的5`端均采用CY5标记。定位探针序列如序列表SEQ ID NO:47所示。具体的序列表如下：
所述的24个SNP位点rs5065、rs12041331、rs5186、rs730012、rs776746、rs2070744、rs12248560、rs4986893、rs4244285、rs1057910、rs1801253、rs2238032、rs2239050、rs2239128、rs2306283、rs4149056、rs671、rs1799752、rs16960228、rs4149601、rs429358、rs7412、rs1065852、rs9923231的检测探针序列如序列表SEQ ID NO:48～SEQ IDNO:95所示；所述阳性探针序列如序列表SEQ ID NO:96～SEQ ID NO:119所示。所述定位参照序列如序列表SEQ ID NO:120所示。
更优选的，所述的检测探针多态性分辨位点采用LNA标记，“+”表示，可提高单碱基的分辨率。为进一步提高检测探针的分辨率探针中有碱基采用次黄嘌呤替代(I)，替代后的探针对特异性更优，同时可以大大提高对捕获效率和减少探针的二级结构的形成。
更优选的，所述的检测探针和阳性探针的5’端采用C12氨基修饰和(dT)15修饰，有效减少了探针与PCR产物在特异性结合时的空间位阻效应。
(二)本实施例的芯片探针序列如下：
芯片的制备工艺如图1所示。如图2所示，图中从上至下1～24号检测区对应的检测位点对应如下：
(三)本实施例的检测试剂盒PCR反应液单人份配置体系如下：
PCR反应液各组分浓度分别为：24个SNP位点rs12041331、rs730012、rs4149056、rs2306283、rs7412、rs429358、rs671、rs9923231、rs12248560、rs4986893、rs4244285、rs5065、rs1799752、rs4149601、rs16960228、rs2239128、rs2239050、rs2238032、rs776746、rs5186、rs1057910、rs1065852、rs2070744、rs1801253前后引物(0.1uM)，Multi BufferMix(1×)，Multi酶系Mix 8.5ul，Solution II 0.25ul，其余用RNase Free-H2O补足；其中Multi Buffer Mix、Multi酶系Mix、Solution II为Multi Enzyme Mix组分，Multi EnzymeMix购自上海生工生物科技有限公司。
(四)本实施例的检测试剂盒芯片制备：
1.探针配制
将探针母液与点样缓冲液进行混合，探针点样终浓度10uM。
2.醛基片清洗
1)把醛基片取出依次放入芯片架中；
2)超声波清洗机装入纯化水达到水位线，再放入已装好芯片的芯片架，浸泡10分钟；
3)浸泡10分钟后，打开超声波清洗机超声清洗5分钟；
4)清洗结束后，取出芯片依次放入玻片离心机中进行离心干燥。
3.芯片点制
保证相对湿度为45％～65％，从2～8℃冰箱内取出384孔板，对384孔板加样，各个位点加样量60μl，将384孔板放入点样仪384孔板槽内。点样仪设置进行点样，点制矩阵示意图如图3所示。
4.芯片水合
温度：37℃；湿度：65％～75％；水合时间：12小时。
5.芯片后处理
将芯片盒内水合好的芯片取出，依次放入24格芯片架内。将芯片架放入0.25％SDS溶液中，5min后取出，再放入纯化水中，上下涮洗10次，以3500rpm离心30s甩干。芯片贴膜和包装。
实施例2、检测试剂盒检测
本发明中采用的仪器如下：扩增仪、杂交仪、GenePix 4100A微阵列基因芯片扫描仪。(1)试剂准备(试剂准备室)
提前将试剂取出，并将PCR反应液涡旋振荡15秒，低速离心待用。确定反应数N，N＝待检样本数(n)+质控品数(1)+空白对照。建议每次PCR实验同时进行阳性对照、空白对照分析。然后将反应液按46μL/管分装至PCR反应管中。
(2)加样检测(样本制备间)
将EDTA抗凝全血、阳性对照和空白对照按4μL加样量加入到PCR反应管中，盖紧管盖，低速离心15秒将管壁上的液体全部甩至管底，然后立即进行PCR扩增反应。
(3)PCR扩增(扩增间)
采用PCR仪进行扩增，反应体系为50μL，反应条件为：预变性95℃，10min；变性95℃，15s，退火60℃，20s，延伸72℃，30s，扩增35个循环；最终延伸72℃，7min；保存4℃∞。
(4)芯片杂交检测
1)将PCR产物及杂交液等比例混匀后在95℃条件下变性10min后，立即置于冰浴中5min；
2)取变性的扩增产物10μl，小心加入基因芯片反应区中，使其分布均匀；
3)将基因芯片平置于杂交盒中，42℃杂交1小时；
4)将基因芯片在SSC洗涤液中漂洗。取出基因芯片，甩去残留在基因芯片上的液体，室温晾干；
5)采用激光共聚焦基因芯片扫描仪对基因芯片进行扫描，并采用信号分析软件进行分析。
(5)结果判读
有效性判定：本检测试剂盒定位参照有信号，空白参照无信号；无模板对照的不通过，阳性对照品的检出结果为杂合型。
最后有必要在此说明的是：以上实施例只用于对本发明的技术方案作进一步详细地说明，不能理解为对本发明保护范围的限制，本领域的技术人员根据本发明的上述内容作出的一些非本质的改进和调整均属于本发明的保护范围。
序列表
<110> 上海普然生物科技有限公司
<120> 一种同时检测心脑血管药物代谢能力基因的引物探针系统及试剂盒
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<212> DNA
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<212> DNA
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<212> DNA
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<212> DNA
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<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 70
caacgctggg ctgcacgcta ctcaccaccc tgccactgc 39
<210> 71
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 71
aatgctagga ttataggcgt gagccaacct ggccaatgg 39
<210> 72
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 72
gtcctccctg gctgttatct tcggtaaaag agaacaca 38
<210> 73
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 73
tcccttctgc tgtctcactt ccgtcttact ctctgctt 38
<210> 74
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 74
cttcactacc aaatgagcct tagctttcag aattgaag 38
<210> 75
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 75
caggggttgc caggaacagc ctggagaccg ggaacaga 38
<210> 76
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 76
gctcttttgt ctttcagtat ctcttgtttg gaccacat 38
<210> 77
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 77
tcaagctctt ccctggccgg ctgagcctca gccctagt 38
<210> 78
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 78
ttgtgtcttc tgttctcaaa gcatgatgta agagataa 38
<210> 79
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 79
tgaaaacatc aggattgtaa gcactggatc caggtaag 38
<210> 80
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 80
actatcattg attatttccc gggaataaca aattactt 38
<210> 81
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 81
cacgaggtcc agagatacct tgactcccca ccagcctg 38
<210> 82
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 82
gacttccgca aggccttcca gggatctgct gcgcgcgca 39
<210> 83
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 83
tggaccagaa aaggccactg tgggcttggg ggtcatgt 38
<210> 84
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 84
atgtctggca aggggctaat tagcaacgtc ttctgaat 38
<210> 85
<211> 38
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 85
ctccagagtc cctgggagcc ccgcctctca ctgcatgt 38
<210> 86
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 86
gtggataagg tcgatgttga atttttgaat cgatattag 39
<210> 87
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 87
aggaatctgg gtcatacatg tggatgcgtt catgggtaa 39
<210> 88
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 88
gggctgcagg catacactga agtgctgtga gtgtgggac 39
<210> 89
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 89
aattggcgaa accacataaa agtgtatagg cagcaggtc 39
<210> 90
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 90
cagctgtggg ctggattctg tattcacacg taaaggaga 39
<210> 91
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 91
gagacgtctc gcatttgagc aggtctcggt aagactttg 39
<210> 92
<211> 40
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 92
gcgatgccga tgacctgcag aagcggcagt gtaccaggcc 40
<210> 93
<211> 40
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 93
aagcggctcc tccgcgatgc cgatgacaga agcgcctggc 40
<210> 94
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 94
caacgctggg ctgcacgcta cccaccaccc tgccactgc 39
<210> 95
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 95
aatgctagga ttataggcgt gagccaaccc ggccaatgg 39
<210> 96
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 96
gagcctcttg cagtctgtcc cta 23
<210> 97
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 97
gacttcccag agaagctgga agg 23
<210> 98
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 98
tgtggactga accgactttt cta 23
<210> 99
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 99
ggaagggctt ccgcagagga gggt 24
<210> 100
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 100
aacgaatgct ctactgtcat ttct 24
<210> 101
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 101
gaactgtagt ttccctagtc cccc 24
<210> 102
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 102
tcagaagacc tcagctcaaa tccc 24
<210> 103
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 103
accacttaca gtcttttttt ctgg 24
<210> 104
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 104
tatggaaagt gatattttgg agaa 24
<210> 105
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 105
atgcaagaca ggagccacat gcc 23
<210> 106
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 106
aactcggcct tcaaccccat ca 22
<210> 107
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 107
agcttccctg gaagaccctg tg 22
<210> 108
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 108
aaaacggaca tctgtatagg ca 22
<210> 109
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 109
gtgtgccttg cctgcctgtt ccct 24
<210> 110
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 110
acatcactga attaaacatt ttgc 24
<210> 111
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 111
aacactctct tatctacata ggtt 24
<210> 112
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 112
ggctacaaga tgtcggggag tgg 23
<210> 113
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 113
tcagagctgg aataaaattg gcg 23
<210> 114
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 114
gcgacattaa gatcaagatc tgc 23
<210> 115
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 115
aatgcactaa acctttaata ttg 23
<210> 116
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 116
gcctcccacc tgcgcaagct gcgt 24
<210> 117
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 117
ctcccacctg cgcaagctgc gta 23
<210> 118
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 118
gccatcttcc tgctcctggt gga 23
<210> 119
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 119
agtgatccac ccacctcggc ctc 23
<210> 120
<211> 18
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 120
cattcaacag ttacgact 18
Claims (1)
1.一种同时检测心脑血管药物代谢能力基因的试剂盒，其特征在于，所述试剂盒同时检测以下24个SNP位点：rs5065、rs12041331、rs5186、rs730012、rs776746、rs2070744、rs12248560、rs4986893、rs4244285、rs1057910、rs1801253、rs2238032、rs2239050、rs2239128、rs2306283、rs4149056、rs671、rs1799752、rs16960228、rs4149601、rs429358、rs7412、rs1065852和rs9923231的基因多态性，扩增所述SNP位点的扩增序列为如SEQ IDNO:1~ SEQ ID NO:23所示的上游引物序列和如SEQ ID NO:24~ SEQ ID NO:46所示的下游引物序列，所述定位探针序列如SEQ ID NO:47所示，所述阳性探针序列如SEQ ID NO:96~SEQ ID NO:119所示，所述定位探针和下游引物序列的5`端均采用CY5标记；所述定位参照序列为NH2-（dT）15-CATTCAACAGTTACGACT；所述24个SNP位点的检测探针序列如下：
其中所述检测探针多态性分辨位点采用LNA标记，用“+”表示，所述检测探针和阳性探针的5’端采用C12氨基修饰和（dT）15修饰，其中“I”表示次黄嘌呤。
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| CN108004312A (zh) * | 2017-12-20 | 2018-05-08 | 德诺杰亿（北京）生物科技有限公司 | 检测心血管疾病相关基因多态性的引物组、试剂盒及方法 |
| CN109402251A (zh) * | 2018-12-14 | 2019-03-01 | 武汉康昕瑞基因健康科技有限公司 | 心脑血管疾病药物相关基因分型检测引物组、试剂盒及检测方法 |
| CN109868314A (zh) * | 2018-12-29 | 2019-06-11 | 浙江迪谱诊断技术有限公司 | 一种检测心血管疾病用药基因的试剂盒及检测方法 |
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