CN112342293B - 家系致病性冠心病易感基因及其体外检测的试剂和应用 - Google Patents
家系致病性冠心病易感基因及其体外检测的试剂和应用 Download PDFInfo
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Abstract
本发明涉及家系致病性冠心病检测技术领域，是一种家系致病性冠心病易感基因及其体外检测的试剂和应用，该家系致病性冠心病易感基因为发生突变的SelS基因，SelS基因的核苷酸序列如序列表SEQ ID NO:1所示，SelS基因发生突变时，其核苷酸序列表第2269位发生可剪切突变。本发明公开了基于冠心病易感基因SelS基因可剪切突变位点而开发的冠心病筛查试剂盒，检测精度和灵敏度高，利用本发明所述试剂盒可以进行冠心病的预测与辅助诊断，对于冠心病的防治具有重要的意义。
Description
技术领域
本发明涉及家系致病性冠心病检测技术领域，是一种家系致病性冠心病易感基因及其体外检测的试剂，还包括所述试剂在制备用于体外检测家系致病性冠心病易感基因的试剂盒中的应用。
背景技术
冠心病(CoronaryArteryDisease，CAD)是指冠状动脉血管发生动脉粥样硬化病变，引起血管腔狭窄、阻塞或冠状动脉血管功能性改变即冠状动脉痉挛，造成心肌缺血、缺氧或坏死而导致的心脏病，又称冠状动脉粥样硬化性心脏病。CAD是一种复杂疾病，对CAD病因学、早期诊断、治疗及预防等方面的研究是目前的前沿问题。研究表明家族史是CAD的独立危险因素，遗传变异被认为是该疾病发生的直接诱因之一，因而，亟需从遗传变异角度开展CAD易感基因的研究。
SelS基因是CAD的主要易感基因已被广泛证实，但SelS基因与CAD具体作用机制尚不十分明确，在SelS基因中，任何一个碱基片段、编码蛋白的突变都会影响其在CAD疾病中的生物过程，对SelS基因致病位点或片段的精确定位有利于冠心病防治药物的筛选和研发。而新近研究表明在导致包括冠心病在内的多种人类疾病的突变中，超过90％不在蛋白质编码范围之内。作为基因调控的关键步骤之一,可变剪接广泛存在于真核生物，并参与各种生物过程,包括细胞分化凋亡和细胞程序性死亡，它也在成熟的组织和细胞压力起着重要的作用，多项研究表明，可变剪接异常可导致许多疾病。因此，研究选择性剪接事件的功能对于理解生物系统的多样性和人类疾病的分子生物学机制至关重要。
发明内容
本发明提供了一种家系致病性冠心病易感基因及其体外检测的试剂和应用，克服了上述现有技术之不足，发明人通过对一个冠心病家系进行了全外显子测序及家系连锁分析，发现了SelS基因的一个可剪切突变(IVS3+3A>T)，人群中验证后，通过构建细胞模型阐明了该突变对于蛋白翻译产生的影响，为冠心病的防治研究提供了参考。鉴于此，本发明以此突变位点为检测靶点，设计引物，开发了一种SelS基因家系突变检测试剂盒，该试剂盒检测精确，灵敏度高，对冠心病的诊断与防治具有重要的指导意义。
本发明的技术方案之一是通过以下措施来实现的：一种家系致病性冠心病易感基因，该易感基因为发生突变的SelS基因，SelS基因的核苷酸序列如序列表SEQ ID NO:1所示，SelS基因发生突变时，其核苷酸序列表第2269位发生可剪切突变。
本发明的技术方案之二是通过以下措施来实现的：一种体外检测家系致病性冠心病易感基因的试剂，试剂包括引物，引物包括上游引物和下游引物，
上游引物的核苷酸序列：TATCCCAGGCTTTATGAATCGG；
下游引物的核苷酸序列：CAGAACAAACTTCACCCTCGTT。
本发明的技术方案之三是通过以下措施来实现的：一种技术方案之二所述试剂在制备用于体外检测家系致病性冠心病易感基因的试剂盒中的应用。
下面是对上述发明技术方案之三的进一步优化或/和改进：
上述试剂盒为聚合酶链式反应与限制性片段长度多态性分析相结合的试剂或聚合酶链式反应与直接测序法相结合的试剂盒。
上述试剂盒中包含PCR扩增酶及相应缓冲液。
本发明公开了基于冠心病易感基因SelS基因可剪切突变位点而开发的冠心病筛查试剂盒，检测精度和灵敏度高，利用本发明的试剂盒可以进行冠心病的预测与辅助诊断，对于冠心病的防治具有重要的意义。
附图说明
附图1为冠心病家系图谱及测序结果。
附图2为Pspl3SELENOSWT、MUT酶切电泳图，图2中，1：质粒；2:XhoI-NheI双酶切后的质粒；M:DNAMarker。
附图3-1为Pspl3–SELENOS-WT(野生型)的质粒测序图谱。
附图3-2为Pspl3–SELENOS-Mut(突变型)的质粒测序图谱。
附图4为Pspl3、Pspl3-mut和Pspl3-wt转染产物的质粒凝胶电泳和测序结果。
具体实施方式
本发明不受下述实施例的限制，可根据本发明的技术方案与实际情况来确定具体的实施方式。
除非特别说明，本发明中的％均为质量百分数；除非特别说明，制备过程在常温、常压状态下进行；除非特别说明，本发明中采用的试剂、方法和设备为本技术领域常规试剂、方法和设备；除非特别说明，本发明中采用的试验条件为本技术领域常规试验条件；除非特别说明，本发明中所用试剂均为市购；除非特别说明，本发明中的水为去离子水。
下面结合实施例对本发明作进一步描述：
实施例1：一种体外检测冠心病易感基因(SelS基因)突变的试剂盒，含有SelS基因的待测样品可以从来自试验者的血液获得。
实施例2：一种SelS基因家系突变检测试剂盒，所述试剂盒中包括引物、探针及扩增试剂：
所述引物序列如下：
上游引物(其核苷酸序列如序列表SEQ ID NO:2所示)：TATCCCAGGCTTTATGAATCGG；
下游引物其核苷酸序列如序列表SEQ ID NO:3所示)：CAGAACAAACTTCACCCTCGTT。
目前关于复杂性状疾病的研究有两种策略，连锁分析的方法和病例-对照研究。由于冠心病是一种多因素疾病，是由遗传和环境因素共同作用的结果，因而采用连锁分析较为困难。从遗传的角度分析，冠心病/心肌梗死属于复杂遗传性状疾病。对该疾病的发生起作用的因素中，除环境因素外，起作用的基因总数估计多达上百条，各个基因之间又存在着相互作用，基因又和环境之间存在相互作用。冠心病的发生具有一定的家族聚集性，但更多的是一些散发病例。所以本发明采用病例对照研究。所述病例对照研究就是在采用随机挑选的两组人群中(病例和对照)比较某等位基因的频率。
实施例3：家系致病性冠心病易感基因(SelS基因)突变位点筛查
在临床工作中，收集了一个患有遗传性冠心病的家庭(图1)。所有患病个体均存活，并接受了药物治疗。该家系为中国汉族，经一中年成年男性因不稳定型心绞痛，入院就诊询问家族史时发现。通过对家族成员进行相关检查，获得了所有亲属的详细临床资料，最终确诊6人患有冠心病，11人无冠心病。对所有血液进行了DNA提取后，通过全外显子测序及生物信息学分析后，确定了该家系的冠心病致病SelS基因一个可剪切突变(IVS3+3A>T)，即核苷酸序列表(SEQ ID NO:1)第2269位发生可剪切突变。
实施例4：SelS基因可剪切突变人群验证
本研究选取2015年1月-2017年12月在新疆医科大学第一附属医院心内科住院CAD患者和无CAD的对照受试者作为验证人群进行研究。
纳入标准：CAD组：按照我院经皮冠状动脉造影证实至少有一支冠状动脉血管狭窄≥50％且无其他方面的心脏病疾患。对照组：选择2015年至2018年新疆地区人群慢性病调查中无任何心脏疾病者，两组在纳入研究前签署知情同意书。
剔除标准：CAD组：临床资料不全者及同时合并以下疾病之一者予以剔除。(如：主动脉夹层、风湿性心脏病、先天性心脏病者、多脏器功能衰竭以及具有精神障碍不能配合者)。
对照组：1、2015年至2018年新疆地区人群慢性病调查中证实患有任何心脏疾病者2、有心血管病家族史者3、吸毒者4、精神障碍者，予以剔除。
采用聚合酶链反应-直接测序的方法检测本发明位点的多态性。
方法：PCR反应体系(20μl)：基因组模板DNA50ng(来自血液标本，按照常规方法将白细胞内的DNA用苯酚-氛仿法或者用盐析法提取出来即可)
最终通过576例CAD与452例进行人群验证。我们并未在人群中发现该位点的突变，证实该位点是一个罕见突变。人群验证基线资料见表1。
反应条件如下表所示：
反应体系如下表：
| ddH2O | 8.0μl |
| 2*TaqPCRmastermix | 10.0μl |
| PrimerF(10nM) | 0.5μl |
| PrimerR(10nM) | 0.5μl |
| DNA | 1.0μl |
| 20.0μl |
实施例5：SelS基因可剪切突变功能研究
Minigene：通指包含一个或者几个外显子以及周围调控区得序列片段。常用来研究不同的蛋白对剪切的调控功能。选择目标的外显子和内含子并进行PCR扩增，把得到的序列片段转染到多种不同的细胞中，通过杂交技术对得到的外显子移除和插入事件进行定量分析为明确该可剪切突变在细胞中的真实剪切情况。我们构建了Pspl3–SELENOS-WT(野生型)和Pspl3–SELENOS-Mut(突变型)质粒。Pspl3-SELENOSWT、Pspl3-SELENOSMut测序结果与参比序列比对一致，质粒构建成功(见图2、3)。
实施例6SelS基因可剪切突变功能研究
将Pspl3-SELENOS野生与突变型质粒转染后Hek293T细胞后，进行采用微小基因分析技术。通过逆转录PCR扩增转录产物，并进行测序分析，确定该突变是否导致内含子剪接异常及其真正的剪接模式。从测序结果可知，c.318+3A(IVS3+3T)发生突变后，内含子剪切的位置发生了变化，向下多剪切了143个碱基，将对蛋白的翻译产生显著影响(见图4)。
以上技术特征构成了本发明的实施例，其具有较强的适应性和实施效果，可根据实际需要增减非必要的技术特征，来满足不同情况的需求。
表1病例对照组基线
| 基线 | 对照组(452例) | 冠心病(576例) | 卡方/t检验 | P值 |
| 年龄 | 54.00(11.70) | 59.00(11.12) | 6.989 | <0.001 |
| 男性比例 | 229(50.7) | 427(74.1) | 60.485 | <0.001 |
| 吸烟 | 122(33.3) | 289(65.8) | 84.357 | <0.001 |
| 饮酒 | 112(24.8) | 201(34.9) | 12.241 | <0.001 |
| 家族史 | 8(1.8) | 20(3.5) | 2.77 | 0.096 |
| 高血压 | 166(36.7) | 323(56.1) | 38.023 | <0.001 |
| 糖尿病 | 49(10.8) | 162(28.1) | 46.382 | <0.001 |
| 甘油三酯 | 1.77(1.36) | 1.99(1.67) | 2.165 | 0.031 |
| 胆固醇 | 4.05(1.07) | 3.70(1.19) | 4.835 | <0.001 |
| 高密度脂蛋白 | 1.16(0.36) | 1.06(0.71) | 2.634 | 0.009 |
| 低密度脂蛋白 | 2.61(0.83) | 2.45(3.26) | 0.986 | 0.324 |
序列表
<110>新疆医科大学第一附属医院
<120>家系致病性冠心病易感基因及其体外检测的试剂和应用
<130>
<160>3
<170>
<210> 1
<211> 6612
<212>DNA
<213> 人类（Homo sapiens）
<400> 1
gaccacgacc gcgtccgggc gcgccacgtg tgcgtaggag cgcaccggaa gcgcccggct 60
ggaggcgcgg cggcagggct gggcggcggc ggcggcggcg gtcatggaac gccaagagga 120
gtctctgtcc gcgcggccgg ccctggagac cgaggggctg cgcttcctgc acaccacggg 180
tgagtcgttg cggggcagcc gggcgcgcgc cgccactttt gcgacgcgca gccatgatgg 240
gtgggtcgtc cgccgctgca ccgggcgccg gagcctggga ggcctgggaa cggtcgggcg 300
ttggcgctta cgcggacctt gggcagcagg cccggacctt gcgcggaggc ttctcgggag 360
ccgcacttcc ctgggcggct cggctgtccc ttgtttgcgc aagtcttttt tgcgaaccaa 420
gcccttcctg tggtagttac tggggtcact cggccgttgg cgtttgcctc tgggacccgt 480
cccacacagc cccatacaca ctcctgactc cccgcgctgt cacccctttc tatgtggctc 540
tgaaaggcct ttgccttcct gattcagatt agttgctctt cattcttcaa aacccagttg 600
ctgtgccctc cacactctaa ctgcccccga ctccccagat ggttgggaag tctcacttct 660
cagtgatccc tgaattgtcg cacttcttga gttcgtgttt taacgatcta cttaggaggc 720
tttttcctca gcctagacca tgaaggcttt gagggcagga gttacacttt gtgtttgttg 780
agtcttatgg aaaggtcaac tagtagtgtc atttttagtt ttttgaaaac tgtttttctt 840
ttcagtgggc tccctgctgg ccacctatgg ctggtacatc gtcttcagct gcatccttct 900
ctacgtggtc tttcagaagc tttccgcccg gctaagagcc ttgaggcaga ggcagctgga 960
ccgagctgcg gctgctgtgg gttagtgcct gataaccgaa atgaaagcgg tggttttgca 1020
cctcctttat attaagagtt agtctcttag taaaagtaag aggggccaca caggaagacc 1080
ctgtctctat ttaaaaaaaa aaaaaatagc cgggagtggc ggcacgcacc tgtagtccca 1140
gctgctcagg aggctgaggc gggataatca cttgagtcca gggagtcaaa gctgcagtgg 1200
gctatgctcg ggccacacta cactccagcc tgggcaattg attgagacct tgtctttaaa 1260
aaaaaaaaaa aaaaaaaagt aggaagtata tggttctcgg tggggcgcgg tggctcacac 1320
ctgtaatccc agcactttgg gaagccgagg caggaggatg acttgaggtc aggggttcga 1380
gaacagcctg gccaacatgg tgaaaccctg tctctactaa aaatacaaat attagtgggg 1440
cgtggtgacg ggcacctgta atcccagcta ttagggtggc tgaggcagga gaaatcgctt 1500
gaacctggga gctggagatt gcagtgagct gagattgtgc cactgcactc cagcctgggc 1560
aacagagtga gactgtcttt tctttctttt tttttttttt ttctatgaga tggagtctag 1620
ccttgttgca aagagcgaga ctctatgagt agacgttatg aatagaaatg agttcatttc 1680
tattcataat gctatttgga aggatttttc ttttctgtag aaacaaatac ttaagaatct 1740
tctgcgctaa ttaagggatg gataatgatt tagaaaactt tatatttcct tggtagtctt 1800
ccaggattct agtcagccta gagactgtgg gtgtcactga ggtatccaag atgtgctctg 1860
tgtggccact atcccaggct ttatgaatcg gaattgctca ggggaactca gaaattggca 1920
tttctaacag atttctggtg atgtagatat ttcgggctaa aatccgtggc tcagcaacag 1980
acccctgccc cctgaagcag taaaatgtat gcagaggggt taggagtact tatgtaaaaa 2040
tatgttgttt cattgtctga tatccatacc tctttatact tttaataata tggacactca 2100
aaagtttcta ttttatattg tacacagtgc tttatctcca tttttttctg acattttaga 2160
acctgatgtt gttgttaaac gacaagaagc tttagcagct gctcgactga aaatgcaaga 2220
agaactaaat gcgcaagttg aaaagcataa ggaaaaactg aaacaagtat gaactggttt 2280
cagtttgaat gtgtgcatag aaattgtctg aggtttagtg gctaacgatg cctgtgtctg 2340
tgttgtctat aagcttctag gaccaggtcc tatcccatta gattcaataa gcatttcagt 2400
tcctaccatg taagtattgg tgatatcaag aagaatacac gattgttagg gaacactaga 2460
tgtgtgaata tattaccatg aaaggtccag agcacaaaag gagggacagg ctggagcagg 2520
gagcatgtga gtgtgtgtgt gcatgtgcct gtgtcttccc cattaccaaa aatgtcctga 2580
caggagtgag tttcagaaga atggagtcag taatcttttt catgaaacat tttgctttct 2640
ttaatagtgt acaaaaacca aagctgctct atgtgagtta aactcacact accagatcac 2700
aacagtttta ttaactaaag aaaacgaggg tgaagtttgt tctgaaagac atttaaatta 2760
agaattatca gagttagctt tgtctttgag agaaatggca gcttctgaat tctttctgta 2820
aaatgtgatt gtttctcagc ttgaagaaga aaaaaggaga cagaagattg aaatgtggga 2880
cagcatgcaa gaaggaaaaa gttacaaagg aaatgcaaag aagccccagg tgactggaga 2940
cctcggccgg ctggcatgcg gtagatgaag attgccaagt agaatgtttt aattgcttct 3000
tacactactg tgtgtgttca aacaggagga agacagtcct gggccttcca cttcatctgt 3060
cctgaaacgg aaatcggaca gaaagccttt gcggggagga ggtaagcacc actgatgtca 3120
aatgttaaca gattttcaac acttacagga tatagttacc ttttaggaac aagattgttt 3180
gtttctttgt ccataaatta agactaattc cttaggattg tgaagattca ataaaggaaa 3240
cagatgcaaa tcacctccta ggtcctcact aagtacttag aaggattgta cttatagtat 3300
tctaacttga tccttctgca gccccgtaga gggagagcta agtagggtga ggaattgtct 3360
gccaatcttc agatgagtgt caaggagctg gaacacagtg gttttggtct ttctggctgg 3420
gaccaccttg tttcttgcaa ataacaagga gtagcagaca gatgctcatc caaagctgct 3480
tcctgtgtgc agcactgccc cggggactct ggatgatgcc acagcagtct gtcttcatcc 3540
catccctgag aatttcaaat ctgggaagat gggactcaca aacgaaaata agcaatcctt 3600
ggtgattctg gctaagagtt gcaagttact gctgaggaag gaaagaacaa acacactaga 3660
acactgtagg aaccaaggcg gaagattttg tatcctccat aggaggagag gggcaccgca 3720
gaggccctga tggtgtcttt gaggactgag gaaagactgg ggcatgggct ccaaggcagc 3780
agggccacag acttggctga ccttaaacgc tgagctgtaa tcccctttgt gtcagaagac 3840
taaacctggc ttgctgtaga gaaggtgatg catctggaaa gaaaatgcta tttttaaatg 3900
gtcctgccgg aagcttattt ttagacacat agaggtgata tttaggagag gaatggaaat 3960
cgtagaagat ggaatgcagg gtgtgcttgc ctgcacggcc tctttcagca tccccagcat 4020
ttctgagctg ggacttttga ctagcctggc tttacaaata aggaaactga ggcacagtgt 4080
ttaattgccc aaagattcca ctataagtaa ggagtaaaag taacatttaa gttctgggtg 4140
gccctagaac cttagcactc aaccaggtta ccagttgtgc actgactttg ggaagctcat 4200
gagggagtgg ggtggttggg ggtagggaag gatacagaag accccgttct gactggtaga 4260
agtgacaagt ttgactcttg atttttttta atctgttttc tgtagcgtga acagccctta 4320
tttgaatgta tgagttttag taagcactgt gataggagga ttcatatact taaatcaggc 4380
cctcttgaga gagttttttg gtgacccttt tgcatgtgtt tcggaggttg ggacaaagaa 4440
gctgaatgac ttttttcccc accagacaat cagttcaaat ggcaatcaca atataaaggt 4500
tttttttttt ttcacatagc taaaaggttt ttttaaatgt cccttaggat ctgtatcttt 4560
gcagtgcttt gcgtgtcact ctcataattt tattgtggat atacaatgtt cccagatttt 4620
cagattttta tcaatactgt tgtgctgctt ttctgtcctc ccaggttata acccgttgtc 4680
tggtgaagga ggcggagctt gctcctggag acctggacgc agaggcccgt catctggcgg 4740
atgaggctaa gaatcttgtt agtgtcactt ttgacattag caagatgaac ccttaaccct 4800
cgattcaatt gccttacgca cgcttttcac agtgactagc caaggggagg tggggttgat 4860
ttctgttcct aactacacct gcatatgtca gggctccagt cagcaaaagg tatagatgtt 4920
gcctctaggc atgaggtcat tggtcacatt ctacttggag acagtgattg cattcattga 4980
tttcatggtt aattgctagt tggtaggtaa aggcctctag atgattagca atcttgataa 5040
aagaggccta gtaatgttct tttgaggtta gaaatccttg ctgctaggac agtctctgtg 5100
acaggttgcg ttgaatgatg tcttccttat caatggtgag cccaccagtg aggattactg 5160
atgtggacag ttgatggggt ttgtttctgt atatttattt ttatgtacag aactttgtaa 5220
aaacgaaact atttaaaaaa caagaataac atttttagca tctttattca aggagattta 5280
tggacttcaa tttgtctatc aaacattaaa tagcttttta ttacaacctc taactattac 5340
ttatttctct atttattatt actcacccaa aataagtctg gcaagtggca gacagctgcc 5400
gtgattttga aagttgacag actccttggg gttcatccac aaaattgggt gcttggagtc 5460
gagttgccga attactggga taaagaaaca tcgtgcctca aggttcctca taggctgacc 5520
aggtttctac gtgacctctc agttattccc ttacgccatg actcatcttt tacgctcttg 5580
tccagaggca ctatcagcag ttttggataa agagccaaac agtaaatatt ttagacttta 5640
caggccacat atgatctctg tcacatagtc ttaggttttt atatttttaa aggattgttt 5700
ttaaaaaaga ttctctgctc atgggctgga tttgggctga atgctgtagt gtgtggaccc 5760
cgtctagatc atcagaagga aaaacattaa ccttcagaat agggggtaat tcgctgcaaa 5820
agtactatag aatggtctca tacataaatt agtcaacatt tgttatctcc tgcaggtaga 5880
atattccgtg gttgcttctt gaccaaggga aatacagtct ggctgtgaga gacttaaaat 5940
ctcttgagga gcgctctgga gaatggctga aggagaggaa acaggagcct tgagcagtgt 6000
aattacaaac aaataggttg gcatagtctt aagtctttga gtctagagag atttgagttt 6060
tgttttgcct gtggaggcaa ttgggggttt caggtcaaag agagggtcag tggaaacaag 6120
ggtgggcctt gtgaggtgtg gggaagccct gggaccctca ctccccttcc agtgtgtaac 6180
tggattggct cccaccagcc cagaagattt acgacgtggg aaatggtata cttggattaa 6240
aatatttcat ccagatttgt ttacatctag agagaacccc ttgtaggtta taaggaaact 6300
ttttaacatt cttccttgaa tatattttct gtagctgaaa atgtttgtga agtggctgtc 6360
aactacacat cgcatgagta gaggggctct gggtcggggt ttcatatgca gcagagcagc 6420
tcccttgctg ccgtccatca agagccctca acacaagagt ttgttataaa tagaaataaa 6480
cgacaaaaag tagaggggta tgtttaagta cactattaga tgtgactgac taccttgaaa 6540
cctgattttt ttccatgggg agaaacagtg cttgaattct gcatgccccc cttgcatcta 6600
gcaccttttg gt 6612
<210> 2
<211>22
<212> DNA
<213> 人工序列
<400> 1
tatcccaggc tttatgaatc gg 22
<210> 3
<211>22
<212> DNA
<213> 人工序列
<400> 1
cagaacaaac ttcaccctcg tt 22
Claims (4)
1.一种体外检测家系致病性冠心病易感基因的试剂，其特征在于所述易感基因为发生突变的SelS基因，SelS基因的核苷酸序列如序列表SEQ ID NO:1所示，SelS基因发生突变时，其核苷酸序列第2269位发生可剪切突变；试剂包括引物，引物包括上游引物和下游引物，
上游引物的核苷酸序列：TATCCCAGGCTTTATGAATCGG；
下游引物的核苷酸序列：CAGAACAAACTTCACCCTCGTT。
2.一种根据权利要求1所述的试剂在制备用于体外检测家系致病性冠心病易感基因的试剂盒中的应用。
3.根据权利要求2所述的应用，其特征在于试剂盒为聚合酶链式反应与限制性片段长度多态性分析相结合的试剂或聚合酶链式反应与直接测序法相结合的试剂盒。
4.根据权利要求2或3所述的应用，其特征在于试剂盒中包含PCR扩增酶及相应缓冲液。
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| CN104513828A (zh) * | 2014-11-25 | 2015-04-15 | 新疆医科大学第一附属医院 | 冠心病相关基因即cyp17a1基因及其体外检测的试剂、制剂或试剂盒、应用 |
| CN108004313A (zh) * | 2017-12-20 | 2018-05-08 | 新疆医科大学第附属医院 | 早发冠心病致病基因及其体外检测的试剂、制剂或试剂盒和应用 |
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| CN104513828A (zh) * | 2014-11-25 | 2015-04-15 | 新疆医科大学第一附属医院 | 冠心病相关基因即cyp17a1基因及其体外检测的试剂、制剂或试剂盒、应用 |
| CN108004313A (zh) * | 2017-12-20 | 2018-05-08 | 新疆医科大学第附属医院 | 早发冠心病致病基因及其体外检测的试剂、制剂或试剂盒和应用 |
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