WO2023142619A1 - 一种扩张型心肌病基因检测标志物及其应用 - Google Patents
一种扩张型心肌病基因检测标志物及其应用 Download PDFInfo
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Definitions
- the invention belongs to the field of molecular biology, and in particular relates to a dilated cardiomyopathy gene detection marker and application thereof.
- Dilated cardiomyopathy is a serious myocardial disease characterized by ventricular dilatation and systolic dysfunction leading to progressive heart failure. Dilated cardiomyopathy is the leading cause of heart failure, accounting for 40-50% of all heart failure causes. The incidence of cardiomyopathy, especially dilated cardiomyopathy, is on the rise, and the prognosis is extremely poor, with a 5-year mortality rate greater than 50.0%. Dilated cardiomyopathy can be divided into two types: familial inheritance and non-familial inheritance, and about 30-50% of dilated cardiomyopathy patients are familial.
- the main inheritance mode of dilated cardiomyopathy is autosomal dominant inheritance, accompanied by X-linked, autosomal recessive inheritance and mitochondrial inheritance.
- Related mutant genes mainly encode cellular components such as sarcomere, cytoskeleton, desmosome and nuclear membrane.
- sarcomere cytoskeleton
- desmosome cytoskeleton
- nuclear membrane Currently, more than 100 genes related to dilated cardiomyopathy have been reported.
- the traditional methods for the differential diagnosis of dilated cardiomyopathy mainly rely on imaging examination methods and the investigation of the patient's medical history. Due to the difficulty in distinguishing the clinical features of dilated cardiomyopathy from myocarditis, arrhythmic cardiomyopathy, and patients with advanced heart failure in terms of cardiac structure and function, and the difficulty in obtaining patient history, there is great uncertainty in the clinical diagnosis of dilated cardiomyopathy . With the continuous maturity of next-generation sequencing technology, the diagnosis rate of dilated cardiomyopathy has increased significantly. At the same time, the reduction in the cost of sequencing has enabled the wider promotion of genetic testing for clinical diseases.
- Chinese patent 201910824992.2 discloses a base site variation that affects the auxiliary diagnosis and clinical intervention of human dilated cardiomyopathy and its application.
- the mutation site of the base site variation that affects the auxiliary diagnosis and prognosis of human dilated cardiomyopathy corresponds to the 44610-2 nucleotide site A>C in the coding region of the human TTN gene, named c.44610-2A >C.
- the invention also provides a primer for identifying the above-mentioned base site variation, discloses the use of identifying the above-mentioned base site variation in the auxiliary diagnosis and clinical intervention of human dilated cardiomyopathy, and the identification of the base site Variant approach for prenatal diagnosis of human dilated cardiomyopathy. It provides a new therapeutic target for the prevention and treatment of dilated cardiomyopathy, and provides a powerful molecular biology tool for the early diagnosis and prognosis of dilated cardiomyopathy.
- Chinese patent 201910315496.4 discloses dilated cardiomyopathy pathogenic gene TTN c.75250C>T mutation and its application.
- Next-generation high-throughput sequencing technology was used to detect gene mutations in the exon and exon regions of the family with dilated cardiomyopathy, and a nonsense mutation (p.R25084X) in exon 326 of the TTN gene (NM_001267550) was found to be associated with dilated cardiomyopathy related.
- Sanger sequencing further verified that the TTN gene c.75250C>T mutation was associated with dilated cardiomyopathy.
- the discovery of this heterozygous mutation provides a basis for analyzing the pathogenic mechanism of dilated cardiomyopathy, and also provides a new direction for the clinical diagnosis and treatment of dilated cardiomyopathy.
- the present invention provides a mutant gene and its new use, which is a homozygous mutation of the human SGCB gene c.243+6A>T site, and the use of the mutation as a molecular marker for dilated cardiomyopathy .
- it provides the use of related reagents for detecting homozygous mutation of human SGCB gene c.243+6A>T site in preparing a detection kit for dilated cardiomyopathy.
- the homozygous mutation of the c.243+6A>T site of SGCB gene is applied to the preparation of dilated cardiomyopathy gene detection kit, which can achieve the purpose of clinical auxiliary diagnosis of dilated cardiomyopathy. Genetic screening is provided for families with pathogenic variants, making it possible to achieve eugenics.
- the present invention provides an application of a gene mutation in the preparation of a detection kit for dilated cardiomyopathy.
- the gene mutation is the pure and mutation of SGCB c.243+6A>T site.
- the meaning of the mutation is that a homogeneous mutation of A>T occurs at the 6th base downstream of the second exon of the human SGCB gene.
- the mutation site is a specific site for East Asian population.
- the present invention provides the application of a reagent for detecting the SGCB c.243+6A>T mutation site in the preparation of a dilated cardiomyopathy detection kit.
- the kit is a gene detection kit.
- the reagents mentioned are sequencing reagents.
- the sequencing reagents include but not limited to Sanger sequencing, cycle array sequencing by synthesis, direct sequencing or fluorescent quantitative PCR.
- the purpose of the sequencing is to detect whether the SGCB c.243+6A>T mutation site occurs.
- the sequencing reagent is a Sanger sequencing reagent, and the sequencing reagent includes primers, and the primers are SEQ ID NO.1-2; wherein SEQ ID NO.1 is an upstream primer, and SEQ ID NO.2 is a downstream primer .
- the present invention provides a kit for detecting dilated cardiomyopathy.
- the kit includes reagents for detecting SGCBc.243+6A>T mutation sites.
- the reagents are Sanger sequencing reagents, cycle array sequencing by synthesis reagents, direct sequencing reagents or fluorescent quantitative PCR reagents.
- the reagents include primers for detecting SGCBc.243+6A>T variant sites.
- the primers can be any primers designed according to the general method that can detect the SGCBc.243+6A>T variant site.
- the primers are SEQ ID NO.1-2.
- the kit provided by the invention can assist in clinical diagnosis of dilated cardiomyopathy.
- the present invention provides a preparation method of a kit for detecting dilated cardiomyopathy, characterized in that the preparation method includes synthesizing primers for detecting the SGCBc.243+6A>T mutation site.
- the present invention provides a method for detecting the aforementioned variant sites using Sanger sequencing.
- the primer combination is SEQ ID NO.1-2.
- SEQ ID NO.1 is a forward primer
- SEQ ID NO.2 is a reverse primer
- the method can be detected by the aforementioned gene detection kit.
- the homozygous mutation of the SGCB c.243+6A>T mutation site in the present invention can be used for the detection of dilated cardiomyopathy.
- the application of the SGCB c.243+6A>T variant site in the detection of dilated cardiomyopathy is different from that in the detection of neuromuscular disease.
- Neuromuscular disease is a genetic disease characterized by motor dysfunction and has a large number of causative genes , the number of mutations is also high, and the disease is not strongly associated with dilated cardiomyopathy.
- Figure 1 shows the comparison results of the sequencing results of patients with cardiomyopathy and the human genome hg19.
- Figure 2 is the Sanger sequencing map of the homozygous mutation at the c.243+6A>T site of SGCB.
- Figure 3 shows the sequencing results of non-cardiomyopathy patients (homozygous).
- Figure 4 shows the sequencing results of non-cardiomyopathy patients (heterozygotes).
- Embodiment 1 A kind of dilated cardiomyopathy gene detection kit
- Upstream primer SEQ ID NO.1 Downstream primer SEQ ID NO.2;
- Embodiment 2 A kind of dilated cardiomyopathy gene detection method
- PCR Mix contains the following components: Taq DNA Polymerase, PCR Buffer, Mg 2+ , dNTPs, PCR stabilizers and enhancers, and other components required for conventional PCR.
- Embodiment 3 Mutation site detection effect verification
- the detection effect is verified by a certain number of irrelevant samples.
- the sample included 500 patients without cardiomyopathy and 11 patients with DCM. It was verified by Sanger sequencing that no homozygous mutation (TT) of SGCB c.243+6A>T site was detected in 500 cases of non-cardiomyopathy population, only wild type (AA) and heterozygous mutation (AT) existed, 11 cases All patients with dilated cardiomyopathy carry homozygous mutations at this site (TT).
- TT homozygous mutation of SGCB c.243+6A>T site was detected in 500 cases of non-cardiomyopathy population, only wild type (AA) and heterozygous mutation (AT) existed, 11 cases All patients with dilated cardiomyopathy carry homozygous mutations at this site (TT).
- test results of 500 cases of non-cardiomyopathy population are as follows:
- serial number result serial number result serial number result serial number result serial number result serial number result serial number result serial number result serial number result serial number result serial number result 38 AA homozygous 171 AA homozygous 289 AA homozygous 428 AA homozygous 549 AA homozygous 39 AA homozygous 172 AA homozygous 290 AA homozygous 429 AA homozygous 550 AA homozygous 40 AA homozygous 173 AT heterozygous 291 AA homozygous 430 AA homozygous 551 AA homozygous 42 AA homozygous 176 AA homozygous 292 AA homozygous 432 AA homozygous 552 AA homozygous 43 AA homozygous 177 AA homozygous 293 AA homozygous 433 AA homozygous 553 AA homozygous 44 AA homozygous 178 AA homozygous 294 AA homozygous 434 AA
- Figure 3-4 The detection results of non-cardiomyopathy patients are shown in Figure 3-4.
- Figure 3 shows a case of AA homozygosity (sample 38), and
- Figure 4 shows a case of AT heterozygosity (sample 276).
Abstract
本发明提供一种基因突变在制备扩张型心肌病检测试剂盒中的应用,所述的基因突变为SGCB c.243+6A>T变异位点。SGCB c.243+6A>T位点的纯合突变可用于临床扩张型心肌病的基因诊断或辅助诊断,为携带扩张型心肌病致病变异的家庭提供遗传筛查,实现优生优育提供可能。
Description
本发明属于分子生物学领域,具体涉及一种扩张型心肌病基因检测标志物及其应用。
扩张型心肌病(dilated cardiomyopathy,DCM),是一种严重的心肌疾病,主要临床特征为心室扩张和收缩功能障碍导致进程性的心力衰竭。扩张型心肌病是导致心力衰竭的主要原因,占所有心力衰竭病因中40-50%。心肌病尤其是扩张型心肌病的发病率有上升趋势,并且预后极差,5年病死率大于50.0%。扩张型心肌病可分为家族遗传性和非家族遗传性两种,其中大约30-50%的扩张型心肌病患者属于家族性。扩张型心肌病主要遗传方式为常染色体显性遗传,同时伴有X连锁、常染色体隐性遗传和线粒体遗传模式。相关突变基因主要编码肌节,细胞骨架,桥粒和核膜等细胞组分。目前,已报道的与扩张型心肌病相关的基因超过100个。
对于扩张型心肌病的鉴别诊断传统的方法主要依靠影像学检查手段以及对患者病史的排查。由于扩张型心肌病临床特征与心肌炎、心律失常性心肌病以及晚期心衰患者在心脏结构和功能上难以区分以及患者病史较难获得等原因,扩张型心肌病临床诊断存在很大的不确定性。随着二代测序技术的不断成熟,扩张型心肌病诊断率显著提高。同时测序成本的降低使临床疾病基因检测得到更加广泛的推广。
中国专利201910824992.2公开了一种影响人类扩张型心肌病辅助诊断及临床干预的碱基位点变异及其应用。所述影响人类扩张型心肌病辅助诊断及预后的碱基位点变异的突变位点对应于人类TTN基因编码区第44610-2位核苷酸位点A>C,命名为c.44610-2A>C。该发明还提供了一种用于鉴定上述碱基位点变异的引物,公开了鉴定前述碱基位点变异在人类扩张型心肌病辅助诊断及临床干预中的用途,以及鉴定该碱基位点变异用于人类扩张型心肌病产前诊断的方法。为防治扩张型心肌病提供了新的治疗靶点,为扩张型心肌病的早期诊断及预后判断提供了强有力的分子生物学工具。
中国专利201910315496.4公开了扩张型心肌病致病基因TTN c.75250C>T突变及其应用。采用新一代高通量测序技术检测扩张性心肌病家系全基因组范围内外显子区域基因突变,发现TTN基因(NM_001267550)外显子326存在1个无义突变(p.R25084X)与扩张型心肌病有关。通过Sanger测序进一步验证了TTN基因c.75250C>T突变与扩张型心肌病有关。该 杂合突变的发现为解析扩张型心肌病的致病机制提供了依据,也为扩张型心肌病临床诊断和治疗提供了新的方向。
但扩张型心肌病发病关联的基因较多,为了能够提高检出效果,提供新的突变位点用于检测诊断或治疗扩张型心肌病是十分必要的。
发明内容
为了解决上述问题,本发明提供了一种突变基因及其新用途,它是人类SGCB基因c.243+6A>T位点的纯合变异,以及该变异作为扩张型心肌病的分子标记的用途。同时提供了检测人类SGCB基因c.243+6A>T位点纯合变异的相关试剂在制备扩张型心肌病的检测试剂盒中的用途。本发明将SGCB基因c.243+6A>T位点的纯合变异应用于扩张型心肌病基因检测试剂盒的制备中,可以达到扩张型心肌病临床辅助诊断的目的,为携带扩张型心肌病致病变异的家庭提供遗传筛查,实现优生优育提供可能。
一方面,本发明提供了一种基因突变在制备扩张型心肌病检测试剂盒中的应用。
所述的基因突变为SGCB c.243+6A>T位点的纯和突变。
所述的突变的含义为人类SGCB基因第二外显子下游第6个碱基处,发生A>T的纯和突变。
所述的突变位点是东亚人群特异位点。
序列比对分析表明SGCB c.243+6A>T位点在多物种中保守,表明该位点在不同物种之间可能具有功能保守性。
另一方面,本发明提供了检测SGCB c.243+6A>T变异位点的试剂在制备扩张型心肌病检测试剂盒中的应用。
所述的试剂盒为基因检测试剂盒。
所述的的试剂为测序试剂。
所述的测序试剂用于包括但不限于Sanger测序、循环阵列合成测序、直接测序或荧光定量PCR。
所述的测序的目的为检测SGCB c.243+6A>T变异位点是否发生。
所述的测序试剂为Sanger测序试剂,所述的测序试剂中包括引物,所述的引物为SEQ ID NO.1-2;其中SEQ ID NO.1为上游引物,SEQ ID NO.2为下游引物。
再一方面,本发明提供了一种用于检测扩张型心肌病的试剂盒。
所述的试剂盒中包括用于检测SGCBc.243+6A>T变异位点的试剂。
所述的试剂为Sanger测序试剂、循环阵列合成测序试剂、直接测序试剂或荧光定量PCR 试剂。
所述的试剂包括用于检测SGCBc.243+6A>T变异位点的引物。
所述的引物可以是根据一般方法设计的任意可对SGCBc.243+6A>T变异位点进行检测的引物。
优选地,所述的引物为SEQ ID NO.1-2。
本发明提供的试剂盒可在临床辅助诊断扩张型心肌病。
又一方面,本发明提供了一种用于检测扩张型心肌病的试剂盒的制备方法,其特征在于,所述的制备方法中包括合成检测SGCBc.243+6A>T变异位点的引物。
又一方面,本发明提供了使用Sanger测序法对前述变异位点进行检测的方法。
所述的方法中包括如下步骤:
(1)基因组DNA提取;
(2)使用经设计的引物组合对SGCB基因进行扩增;
(3)PCR产物进行Sanger测序;
(4)测序结果分析。
所述的引物组合为SEQ ID NO.1-2。其中SEQ ID NO.1为正向引物,SEQ ID NO.2为反向引物。
所述的方法可以通过前述的基因检测试剂盒进行检测。
本发明的有益效果:
本发明中SGCB c.243+6A>T变异位点的纯合突变可用于扩张型心肌病的检测。SGCB c.243+6A>T变异位点应用于扩张型心肌病的检测与应用于神经肌肉病的检测不同,神经肌肉病是一种表现为运动功能障碍的遗传性疾病,致病基因数目众多,突变数量也众多,且该疾病与扩张型心肌病关联不大。
图1为心肌病患者测序结果与人的基因组hg19比对结果。
图2为SGCB c.243+6A>T位点的纯合突变Sanger测序图。
图3为非心肌病患者测序结果(纯合子)。
图4为非心肌病患者测序结果(杂合子)。
下面结合具体实施例,对本发明作进一步详细的阐述,下述实施例不用于限制本发明,仅用于说明本发明。以下实施例中所使用的实验方法如无特殊说明,实施例中未注明具体条件的实验方法,通常按照常规条件,下述实施例中所使用的材料、试剂等,如无特殊说明,均可从商业途径得到。
基础实验例变异位点的筛选
对593例样本(包括扩张型心肌病和先天性心脏病对照)进行全外显子组位点关联分析,得到了SGCB c.243+6A>T变异位点在全基因组范围内与扩张型心肌病具有显著相关性,因此将SGCB c.243+6A>T纯和变异作为扩张型心肌病检测标志物。
实施例1一种扩张型心肌病基因检测试剂盒
本实施例的试剂盒中包括以下组分:
(1)上游引物SEQ ID NO.1;下游引物SEQ ID NO.2;
(2)DNA提取试剂;
(3)Taq DNA Polymerase;
(4)PCR Buffer;
(5)Mg
2+;
(6)dNTPs;
(7)PCR稳定剂和增强剂。
实施例2一种扩张型心肌病基因检测方法
(1)基因组DNA提取
对人类心脏组织样本进行全基因组DNA的提取,对DNA的浓度和纯度进行检测。
(2)使用经设计的引物组合对SGCB基因进行扩增
利用已设计好的引物(表1),配置到PCR扩增反应体系(表2)中,放置于PCR反应仪器中,利用扩增程序(表3),将包含突变位点的一段DNA序列从基因组中扩增出来。
表1.引物信息
引物名称 | 引物序列(5’-3’) | |
上游引物 | SGCB-F | SEQ ID NO.1 |
下游引物 | SGCB-R | SEQ ID NO.2 |
表2.PCR扩增反应体系
试剂名称 | 体积(μL) | 试剂供应商 |
PCR Mix(2×) | 10 | 南京诺唯赞生物科技股份有限公司 |
SGCB-F(10um) | 1 | 北京天一辉远生物科技有限公司 |
SGCB-R(10um) | 1 | 北京天一辉远生物科技有限公司 |
基因组DNA | 1 | 本室提取 |
ddH 2O 2 | 补至20 | 本室配置 |
注:PCR Mix中包含如下成分:Taq DNA Polymerase、PCR Buffer、Mg
2+、dNTPs、PCR稳定剂和增强剂等常规PCR所需要的组分。
表3.PCR反应程序
(3)PCR产物进行Sanger测序;将扩增出的PCR产物进行核酸电泳,判断片段大小是否正确,片段大小正确后送公司(北京天一辉远生物科技有限公司)进行Sanger一代测序。
(4)测序结果分析。将测序得到的reads比对到人的基因组hg19上,在比对结果bam文件中,纯合变异个体(扩张型心肌病患者)的chr4:52899591位置显示只有突变碱基T的存在,不存在野生型等位基因(图1)。测序结果分析显示携带SGCB c.243+6A>T位点的纯合突变结果如图2,不携带此位点的纯合突变的个体为非扩张型心肌病患者。
实施例3突变位点检测效果验证
通过一定数量的无关样本进行检测效果的验证。
基于扩张型心肌病发病的罕见性,样本包括500例非心肌病患者和11例扩张型心肌病患者。经Sanger测序验证证实:500例非心肌病人群未检测到SGCB c.243+6A>T位点的纯合突变(TT),仅存在野生型(AA)和杂合突变(AT),11例扩张型心肌病患者全部携带此位点的纯合突变(TT)。
500例非心肌病人群检测结果如下:
编号 | 结果 | 编号 | 结果 | 编号 | 结果 | 编号 | 结果 | 编号 | 结果 |
38 | AA纯合 | 171 | AA纯合 | 289 | AA纯合 | 428 | AA纯合 | 549 | AA纯合 |
39 | AA纯合 | 172 | AA纯合 | 290 | AA纯合 | 429 | AA纯合 | 550 | AA纯合 |
40 | AA纯合 | 173 | AT杂合 | 291 | AA纯合 | 430 | AA纯合 | 551 | AA纯合 |
42 | AA纯合 | 176 | AA纯合 | 292 | AA纯合 | 432 | AA纯合 | 552 | AA纯合 |
43 | AA纯合 | 177 | AA纯合 | 293 | AA纯合 | 433 | AA纯合 | 553 | AA纯合 |
44 | AA纯合 | 178 | AA纯合 | 294 | AA纯合 | 434 | AA纯合 | 554 | AA纯合 |
45 | AA纯合 | 179 | AA纯合 | 295 | AA纯合 | 435 | AA纯合 | 555 | AA纯合 |
46 | AA纯合 | 180 | AA纯合 | 296 | AA纯合 | 437 | AA纯合 | 556 | AT杂合 |
47 | AA纯合 | 181 | AA纯合 | 298 | AA纯合 | 439 | AA纯合 | 557 | AA纯合 |
48 | AA纯合 | 182 | AA纯合 | 299 | AA纯合 | 440 | AA纯合 | 558 | AA纯合 |
49 | AA纯合 | 183 | AA纯合 | 300 | AA纯合 | 441 | AA纯合 | 559 | AT杂合 |
50 | AA纯合 | 184 | AA纯合 | 302 | AA纯合 | 442 | AA纯合 | 560 | AA纯合 |
52 | AA纯合 | 185 | AA纯合 | 303 | AA纯合 | 443 | AA纯合 | 561 | AA纯合 |
53 | AA纯合 | 186 | AA纯合 | 305 | AA纯合 | 444 | AA纯合 | 562 | AA纯合 |
54 | AA纯合 | 187 | AA纯合 | 306 | AA纯合 | 445 | AA纯合 | 563 | AA纯合 |
55 | AA纯合 | 188 | AA纯合 | 307 | AA纯合 | 446 | AA纯合 | 564 | AA纯合 |
56 | AA纯合 | 189 | AA纯合 | 308 | AA纯合 | 447 | AT杂合 | 565 | AA纯合 |
57 | AA纯合 | 190 | AA纯合 | 309 | AA纯合 | 448 | AA纯合 | 567 | AA纯合 |
58 | AA纯合 | 191 | AT杂合 | 310 | AA纯合 | 449 | AA纯合 | 568 | AA纯合 |
59 | AA纯合 | 192 | AA纯合 | 312 | AA纯合 | 450 | AA纯合 | 569 | AA纯合 |
60 | AA纯合 | 193 | AA纯合 | 314 | AA纯合 | 451 | AA纯合 | 570 | AA纯合 |
61 | AA纯合 | 194 | AA纯合 | 315 | AA纯合 | 452 | AA纯合 | 571 | AA纯合 |
62 | AA纯合 | 196 | AA纯合 | 317 | AA纯合 | 453 | AA纯合 | 572 | AA纯合 |
63 | AA纯合 | 197 | AA纯合 | 318 | AA纯合 | 454 | AA纯合 | 573 | AA纯合 |
64 | AA纯合 | 199 | AA纯合 | 331 | AA纯合 | 455 | AA纯合 | 574 | AA纯合 |
66 | AT杂合 | 200 | AA纯合 | 332 | AA纯合 | 456 | AA纯合 | 575 | AA纯合 |
67 | AA纯合 | 201 | AA纯合 | 333 | AA纯合 | 457 | AA纯合 | 576 | AA纯合 |
68 | AA纯合 | 202 | AA纯合 | 334 | AA纯合 | 459 | AA纯合 | 577 | AA纯合 |
69 | AA纯合 | 203 | AA纯合 | 337 | AA纯合 | 460 | AA纯合 | 578 | AA纯合 |
70 | AA纯合 | 204 | AA纯合 | 338 | AA纯合 | 461 | AA纯合 | 579 | AA纯合 |
71 | AA纯合 | 205 | AA纯合 | 339 | AA纯合 | 462 | AA纯合 | 580 | AA纯合 |
73 | AA纯合 | 206 | AA纯合 | 340 | AA纯合 | 464 | AA纯合 | 581 | AA纯合 |
74 | AA纯合 | 207 | AA纯合 | 341 | AT杂合 | 465 | AA纯合 | 582 | AA纯合 |
75 | AA纯合 | 208 | AA纯合 | 342 | AA纯合 | 466 | AA纯合 | 583 | AA纯合 |
76 | AA纯合 | 209 | AA纯合 | 343 | AA纯合 | 467 | AA纯合 | 584 | AA纯合 |
77 | AA纯合 | 210 | AA纯合 | 344 | AA纯合 | 468 | AA纯合 | 585 | AA纯合 |
78 | AA纯合 | 211 | AA纯合 | 346 | AA纯合 | 469 | AA纯合 | 586 | AA纯合 |
79 | AA纯合 | 212 | AA纯合 | 348 | AA纯合 | 470 | AA纯合 | 587 | AA纯合 |
80 | AA纯合 | 213 | AA纯合 | 349 | AA纯合 | 471 | AA纯合 | 588 | AA纯合 |
83 | AA纯合 | 214 | AA纯合 | 351 | AA纯合 | 473 | AA纯合 | 589 | AA纯合 |
85 | AT杂合 | 215 | AA纯合 | 352 | AA纯合 | 474 | AA纯合 | 590 | AA纯合 |
86 | AT杂合 | 216 | AA纯合 | 353 | AA纯合 | 475 | AA纯合 | 591 | AA纯合 |
87 | AA纯合 | 217 | AA纯合 | 355 | AA纯合 | 476 | AA纯合 | 592 | AA纯合 |
88 | AA纯合 | 218 | AA纯合 | 356 | AA纯合 | 477 | AA纯合 | 593 | AT杂合 |
89 | AA纯合 | 219 | AA纯合 | 357 | AA纯合 | 478 | AA纯合 | 596 | AA纯合 |
90 | AA纯合 | 220 | AA纯合 | 358 | AA纯合 | 480 | AA纯合 | 597 | AA纯合 |
91 | AA纯合 | 221 | AA纯合 | 359 | AA纯合 | 481 | AT杂合 | 598 | AA纯合 |
92 | AA纯合 | 222 | AA纯合 | 360 | AA纯合 | 483 | AA纯合 | 599 | AA纯合 |
94 | AA纯合 | 223 | AA纯合 | 362 | AA纯合 | 484 | AA纯合 | 600 | AA纯合 |
96 | AA纯合 | 224 | AA纯合 | 363 | AA纯合 | 486 | AA纯合 | 601 | AA纯合 |
97 | AA纯合 | 225 | AA纯合 | 364 | AA纯合 | 487 | AA纯合 | 602 | AA纯合 |
98 | AA纯合 | 226 | AA纯合 | 365 | AA纯合 | 488 | AA纯合 | 603 | AA纯合 |
99 | AA纯合 | 227 | AA纯合 | 366 | AA纯合 | 489 | AA纯合 | 604 | AA纯合 |
100 | AA纯合 | 228 | AA纯合 | 367 | AA纯合 | 490 | AA纯合 | 605 | AA纯合 |
101 | AA纯合 | 230 | AA纯合 | 368 | AA纯合 | 491 | AA纯合 | 606 | AA纯合 |
102 | AA纯合 | 231 | AA纯合 | 369 | AA纯合 | 492 | AA纯合 | 607 | AA纯合 |
103 | AA纯合 | 232 | AA纯合 | 370 | AA纯合 | 493 | AA纯合 | 608 | AA纯合 |
105 | AA纯合 | 235 | AA纯合 | 371 | AA纯合 | 494 | AA纯合 | 610 | AA纯合 |
110 | AA纯合 | 236 | AA纯合 | 373 | AA纯合 | 495 | AA纯合 | 611 | AA纯合 |
115 | AA纯合 | 237 | AA纯合 | 374 | AA纯合 | 496 | AA纯合 | 612 | AA纯合 |
116 | AA纯合 | 238 | AA纯合 | 375 | AA纯合 | 497 | AA纯合 | 613 | AA纯合 |
118 | AA纯合 | 239 | AA纯合 | 377 | AA纯合 | 498 | AA纯合 | 614 | AA纯合 |
120 | AA纯合 | 241 | AA纯合 | 378 | AA纯合 | 500 | AA纯合 | 616 | AA纯合 |
122 | AA纯合 | 242 | AT杂合 | 379 | AA纯合 | 501 | AA纯合 | 617 | AA纯合 |
123 | AA纯合 | 245 | AA纯合 | 381 | AA纯合 | 502 | AT杂合 | 618 | AA纯合 |
124 | AA纯合 | 246 | AA纯合 | 382 | AA纯合 | 503 | AA纯合 | 619 | AA纯合 |
125 | AA纯合 | 247 | AA纯合 | 383 | AA纯合 | 504 | AA纯合 | 620 | AA纯合 |
126 | AA纯合 | 248 | AA纯合 | 384 | AA纯合 | 505 | AA纯合 | 621 | AA纯合 |
127 | AA纯合 | 249 | AA纯合 | 385 | AA纯合 | 506 | AA纯合 | 622 | AA纯合 |
128 | AA纯合 | 250 | AA纯合 | 386 | AA纯合 | 507 | AA纯合 | 623 | AA纯合 |
130 | AA纯合 | 251 | AA纯合 | 387 | AA纯合 | 508 | AA纯合 | 625 | AT杂合 |
131 | AA纯合 | 253 | AA纯合 | 388 | AA纯合 | 511 | AA纯合 | 626 | AA纯合 |
132 | AA纯合 | 254 | AA纯合 | 389 | AA纯合 | 512 | AA纯合 | 627 | AA纯合 |
133 | AA纯合 | 258 | AA纯合 | 390 | AA纯合 | 513 | AA纯合 | 628 | AA纯合 |
134 | AA纯合 | 259 | AA纯合 | 391 | AA纯合 | 514 | AA纯合 | 629 | AA纯合 |
135 | AA纯合 | 260 | AA纯合 | 392 | AA纯合 | 516 | AA纯合 | 630 | AA纯合 |
136 | AA纯合 | 262 | AA纯合 | 396 | AA纯合 | 517 | AA纯合 | 631 | AA纯合 |
138 | AA纯合 | 265 | AA纯合 | 398 | AA纯合 | 520 | AA纯合 | 632 | AA纯合 |
139 | AA纯合 | 266 | AA纯合 | 399 | AA纯合 | 521 | AA纯合 | 633 | AA纯合 |
140 | AA纯合 | 267 | AA纯合 | 400 | AA纯合 | 522 | AA纯合 | 634 | AA纯合 |
141 | AT杂合 | 268 | AA纯合 | 401 | AA纯合 | 523 | AA纯合 | 635 | AA纯合 |
142 | AA纯合 | 269 | AA纯合 | 402 | AA纯合 | 524 | AA纯合 | 636 | AA纯合 |
143 | AA纯合 | 270 | AA纯合 | 404 | AA纯合 | 525 | AA纯合 | 637 | AA纯合 |
144 | AA纯合 | 271 | AA纯合 | 406 | AA纯合 | 526 | AA纯合 | 638 | AA纯合 |
148 | AA纯合 | 272 | AA纯合 | 407 | AA纯合 | 527 | AA纯合 | 639 | AA纯合 |
149 | AA纯合 | 273 | AA纯合 | 408 | AA纯合 | 528 | AA纯合 | 640 | AA纯合 |
151 | AT杂合 | 274 | AA纯合 | 409 | AA纯合 | 530 | AA纯合 | 641 | AA纯合 |
156 | AA纯合 | 275 | AA纯合 | 410 | AA纯合 | 531 | AA纯合 | 642 | AA纯合 |
157 | AA纯合 | 276 | AT杂合 | 411 | AA纯合 | 532 | AA纯合 | 649 | AA纯合 |
158 | AT杂合 | 277 | AA纯合 | 412 | AA纯合 | 533 | AA纯合 | 650 | AA纯合 |
160 | AA纯合 | 278 | AA纯合 | 413 | AA纯合 | 535 | AA纯合 | 651 | AA纯合 |
161 | AA纯合 | 279 | AA纯合 | 414 | AA纯合 | 536 | AA纯合 | 652 | AA纯合 |
162 | AA纯合 | 280 | AA纯合 | 415 | AA纯合 | 537 | AA纯合 | 653 | AA纯合 |
163 | AA纯合 | 281 | AA纯合 | 416 | AA纯合 | 539 | AA纯合 | 654 | AA纯合 |
164 | AA纯合 | 283 | AA纯合 | 417 | AA纯合 | 541 | AA纯合 | 655 | AA纯合 |
165 | AA纯合 | 284 | AA纯合 | 418 | AA纯合 | 543 | AA纯合 | 659 | AA纯合 |
166 | AA纯合 | 285 | AA纯合 | 419 | AA纯合 | 544 | AA纯合 | 660 | AA纯合 |
167 | AA纯合 | 286 | AA纯合 | 420 | AA纯合 | 545 | AA纯合 | 662 | AA纯合 |
168 | AA纯合 | 287 | AA纯合 | 423 | AT杂合 | 546 | AA纯合 | 663 | AA纯合 |
170 | AA纯合 | 288 | AA纯合 | 425 | AA纯合 | 548 | AA纯合 | 664 | AA纯合 |
非心肌病患者的检测结果见图3-4。其中图3为AA纯合一例(样本38),图4为AT杂合一例(样本276)。
以上说明,SGCB c.243+6A>T位点的纯合突变可用于扩张型心肌病的检测。
Claims (10)
- 一种基因突变在制备扩张型心肌病检测试剂盒中的应用,其特征在于,所述的基因突变为SGCB c.243+6A>T纯合变异。
- 检测SGCB c.243+6A>T变异位点的试剂在制备扩张型心肌病检测试剂盒中的应用。
- 根据权利要求1-2任一项所述的应用,其特征在于,所述的试剂盒为基因检测试剂盒。
- 根据权利要求2所述的应用,其特征在于,所述的的试剂为测序试剂。
- 根据权利要求4所述的应用,其特征在于,所述的测序试剂为Sanger测序试剂、循环阵列合成测序试剂、直接测序试剂或荧光定量PCR试剂。
- 根据权利要求5所述的应用,其特征在于,所述的测序试剂为Sanger测序试剂,所述的测序试剂中包括引物,所述的引物为SEQ ID NO.1-2。
- 一种试剂盒,其特征在于,包括用于检测SGCB c.243+6A>T变异位点的试剂。
- 根据权利要求7所述的试剂盒,其特征在于,所述的试剂为Sanger测序试剂、循环阵列合成测序试剂、直接测序试剂或荧光定量PCR试剂。
- 根据权利要求8所述的试剂盒,其特征在于,包括用于检测SGCB c.243+6A>T变异位点的引物,所述的引物为SEQ ID NO.1-2。
- 一种试剂盒的制备方法,其特征在于,所述的制备方法中包括合成检测SGCBc.243+6A>T变异位点的引物。
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US6201168B1 (en) * | 1999-08-20 | 2001-03-13 | University Of Iowa Research Foundation | Pathogenesis of cardiomyopathy |
US20120030779A1 (en) * | 2007-01-18 | 2012-02-02 | University Of Utah Research Foundtion | Compositions and methods for detecting, treating, or preventing reductive stress |
US20190350955A1 (en) * | 2016-06-30 | 2019-11-21 | Inserm (Institut National De La Sante Et De La Recherche Medicale | Methods and pharmaceutical compositions for the treatment of cardiomyopathies |
US20200261589A1 (en) * | 2017-10-20 | 2020-08-20 | Genethon | Use of a Syncytin for Targeting Drug and Gene Delivery to Regenerate Muscle Tissue |
CN113981066A (zh) * | 2021-11-02 | 2022-01-28 | 百世诺(北京)医疗科技有限公司 | 突变的扩张型心肌病致病基因ttn及其应用 |
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US6201168B1 (en) * | 1999-08-20 | 2001-03-13 | University Of Iowa Research Foundation | Pathogenesis of cardiomyopathy |
US20120030779A1 (en) * | 2007-01-18 | 2012-02-02 | University Of Utah Research Foundtion | Compositions and methods for detecting, treating, or preventing reductive stress |
US20190350955A1 (en) * | 2016-06-30 | 2019-11-21 | Inserm (Institut National De La Sante Et De La Recherche Medicale | Methods and pharmaceutical compositions for the treatment of cardiomyopathies |
US20200261589A1 (en) * | 2017-10-20 | 2020-08-20 | Genethon | Use of a Syncytin for Targeting Drug and Gene Delivery to Regenerate Muscle Tissue |
CN113981066A (zh) * | 2021-11-02 | 2022-01-28 | 百世诺(北京)医疗科技有限公司 | 突变的扩张型心肌病致病基因ttn及其应用 |
CN114507727A (zh) * | 2022-01-29 | 2022-05-17 | 中国医学科学院阜外医院 | 一种扩张型心肌病基因检测标志物及其应用 |
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