CN106399470A - 一种脑海绵状血管瘤突变基因及检测试剂盒 - Google Patents
一种脑海绵状血管瘤突变基因及检测试剂盒 Download PDFInfo
- Publication number
- CN106399470A CN106399470A CN201610503723.2A CN201610503723A CN106399470A CN 106399470 A CN106399470 A CN 106399470A CN 201610503723 A CN201610503723 A CN 201610503723A CN 106399470 A CN106399470 A CN 106399470A
- Authority
- CN
- China
- Prior art keywords
- cerebral cavernous
- cavernous angioma
- mutant gene
- gene
- angioma
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Analytical Chemistry (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Hospice & Palliative Care (AREA)
- Biophysics (AREA)
- Oncology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
本发明公开了一种脑海绵状血管瘤突变基因,所述突变基因的序列如SEQ ID NO.1所示。该突变基因可用作检测脑海绵状血管瘤的生物标志物,还可以用于制备检测脑海绵状血管瘤试剂、试剂盒或生物芯片等。同时,本发明提供了一种检测脑海绵状血管瘤的试剂盒,所述试剂盒中包括如下引物对:上游引物:5’‑GCCCGGCCAGTAAAATGTTT‑3’(SEQ ID NO.1),下游引物:5’‑TTGAATGAAGCAGGGAACGG‑3’(SEQ ID NO.2),该引物对即是检测所述突变基因的引物对。
Description
技术领域
本发明涉及一种脑海绵状血管瘤突变基因及检测试剂盒。
背景技术
脑海绵状血管瘤(Cerebral Cavernous Malformations,CCM)是中枢神经系统隐匿性血管畸形的一种,临床上分为家族性和散发性两大类,目前统计人群发病率约为0.5%,占中枢神经系统血管畸形的10—20%,其中家族性脑海绵状血管瘤约50%。该病的主要临床表现为癫痫(占37-—50%)、脑出血(占20—40%)、非特异性反复头痛(占10—30%)和局灶性中枢神经功能缺损症状(占35—40%)。约9%的患者在10岁前发病,60—70%的患者发病年龄在10—40岁间,有19%左右的患者在40岁后发病,而25%有脑海绵状血管瘤的个体终生无症状,此类患者大多数在因其他疾病做头部影像学检查及尸体解剖时发现。
脑海绵状血管瘤患者临床特点多样而且无特异性,因此CT(Computed Tomography,计算机断层扫描)和MRI(Magnetic Resonance Imaging,磁共振成像)为术前诊断的主要方法,术后的病理检查为金标准。该病CT平扫特征性表现为脑实质中边界清楚的圆形或不规则形高密度或混杂密度灶,增强后70%-94%的病变可有轻、中度增强,典型表现为不均匀的斑点状增强,但是灶周水肿一般不明显。磁共振扫描检查对CCM的诊断具有较高的特异性:在T2WI序列表现混杂信号,周围可有一均匀的低信号环;在T1WI序列多表现为均匀的低或等信号影。病灶周围无明显水肿,增强扫描无明显强化效应,无血管流空效应,多数临床医师认为上述特征性的表现是诊断颅内脑海绵状血管瘤的主要依据,一般DSA检查脑海绵状血管瘤呈阴性,故又称隐匿性血管畸形。脑海绵状血管瘤是由不规则的、大小不等的血管腔隙聚集成一堆的血管组织,其管壁仅为一层扁平血管内皮细胞和少量成纤维细胞,无成形的血管壁,病灶内无神经组织,其病理结构类是于肝窦结构,剖面呈海绵状或蜂窝状。术后的病理检查仍是CCM诊断的金标准。
临床上脑海绵状血管瘤分为家族性脑海绵状血管瘤和散发性脑海绵状血管瘤。家族性脑海绵状血管瘤多发病灶常见,散发性脑海绵状血管瘤多为单发病灶。家族性脑海绵状血管瘤呈常染色体不完全显性遗传,目前被已明确并克隆的致病基因有CCM1(KRIT1,7q11.2-q21),CCM2(MGC4607,7p15-p13)和CCM3(PDCD10,3q26.1)。虽然散发性脑海绵状血管瘤无遗传性,但国外也相继有新发现突变位点的报导。更值得注意的是,目前大多数研究者认为散发性多发病灶脑海绵状血管可能是遗传性病例,来自无症状父母。
CCM1基因含有20个外显子,其中外显子4到外显子19共16个外显子编码合成含有736个氨基酸的Krit1蛋白,因此又称之为KRIT1基因。1982年Hayman等对一个庞大的墨西哥家系进行遗传学研究,证实该病呈常染色体显性遗传后,1995年Dubovsky等对一西班牙裔人脑海绵状血管瘤大家系采用连锁分析和短串联重复序列多态性的方法将CCM1定位到7q11-q22。Gunel等也用连锁分析方法将致病基因CCM1更精细地定位到7q11.2-q21,发现CCM1基因5’端存在8个新的外显子,使得CCM1的完整基因充分被人们所认识。
CCM2基因表达的蛋白产物为MGC4607,所以称为MGC4607基因,被Denier等在2004年对30个无CCM1基因突变的脑海绵状血管瘤家系进行遗传连锁分析,而确定在7p15-p13上,含有10个外显子,编码含有444个氨基酸的MGC460蛋白。
2005年Bergametti等[14]对无CCM1和CCM2突变的20个脑海绵状血管瘤家用18个基因微卫星标记,将CCM3基因位置逐步确定在3q25.2-27,确定CCM3基因就是PCD10基因,含有10个外显子,其中有七个外显子编码形成含有222个氨基酸的PDCD10蛋白。
到目前为止已发现报导了110余个CCM1突变位点,近40个CCM2突变位点,约60个CCM3突变位点(本文所附综述中有详细介绍)。迄今的报道,仅欧美国家有较大样本量的研究,其他地区,比如我国、日本,对脑海绵状血管瘤遗传基因报道多仅是在个别家系发现了新的突变位点。目前国内研究仅报道了CCM1基因上8个突变位点,尚无CCM2、CCM3基因突变的报道。
目前我国对脑海绵状血管瘤基因突变方向的研究较少,多发性脑海绵状血管瘤尤其弥漫性较少见,而较多研究发现散发性多发性脑海绵状血管瘤其实也是遗传病例,而也有散发单发病灶病例为来自无症状父母的遗传病例。
发明内容
本发明的发明人通过调查病例资料和电话随访,收集了2011年-2015年在我院神经内外科住院的汉族脑海绵状血管瘤患者78例,男性43例,女性35例。发病年龄在2-77岁,平均发病年龄45.73±17.45岁。多发病灶3例,单发病灶75例。所有病例经神经内外科及影像科3位副主任或者主任医师诊断,符合周良辅主编第二版《中国现代神经外科学》脑海绵状血管瘤的诊断标准,均具有典型临床症状和影像学特征,其中经术后病理证实28例。通过详系调查患者家属的所有就诊经历、免费MRI检查和测序分析,本研究采集血样的22例患者证实均是散发性脑海绵状血管瘤。通过详系调查患者家属的所有就诊经历,其余56例患者也基本明确为是散发性脑海绵状血管瘤。
抽取22例(3例多发病灶,19例单发病灶)病人和9位可疑家属的外周血3-4ml,放置EDTAK2(或EDTANa4)抗凝管中,均放置在-80℃超低温冰箱保存。选取100位健康人群作为对照组,经知情同意后抽取外周血3-4ml,同样方法保存。样本送至华大基因科技有限公司武汉分公司进行CCM1、CCM2和CCM3基因全场测序。
测序结果发现,多发病病灶病例S29检测到的CCM1基因c.1784delC,导致框移突变,导致KRIT1蛋白在595位氨基酸开始出现错位,并提前出现终止密码,KRIT1蛋白功能缺失。在100位对照组中没有检测CCM1基因c.1784delC,变该变异位点在突变数据库及SNP数据库都没有收录,既往也无文献报道,该突变基因序列如SEQ ID NO.1所示,即为本发明所述的脑海绵状血管瘤突变基因,该突变基因可用作检测脑海绵状血管瘤的生物标志物,还可以用于制备检测脑海绵状血管瘤试剂、试剂盒或生物芯片等。同时,本发明提供了一种检测脑海绵状血管瘤的试剂盒,所述试剂盒中包括如下引物对:上游引物:5’-GCCCGGCCAGTAAAATGTTT-3’(SEQ ID NO.1),下游引物:5’-TTGAATGAAGCAGGGAACGG-3’(SEQ ID NO.2),该引物对即是检测所述突变基因的引物对。另外,本发明还对该突变基因进行了SNP验证:收集100例健康人的血样标本,提取全基因组DNA。然后对CCM1基因13号外显子进行测序分析,发现100例健康人种均没有发现c.1784delC突变,因此认定该突变不是单核苷酸多态性(SNP)。
附图说明
图1为KRIT1基因c.1784delC突变测序图,图中:突变测序结果及正常对照结果(箭头示突变位点);
具体实施方式
对多发病病灶病例S29号标本进行CCM1、CCM2和CCM3基因全场测序,检测到的CCM1基因c.1784delC(图1),检测CCM1基因c.1784delC的引物对为:
5’-GCCCGGCCAGTAAAATGTTT-3’(SEQ ID NO.1),下游引物:
5’-TTGAATGAAGCAGGGAACGG-3’(SEQ ID NO.2),导致框移突变,导致KRIT1蛋白在595位氨基酸开始出现错位,并提前出现终止密码,KRIT1蛋白功能缺失。在100位对照组中没有检测CCM1基因c.1784delC,变该变异位点在突变数据库及SNP数据库都没有收录,既往也无文献报道。
Claims (4)
1.一种脑海绵状血管瘤突变基因,其特征在于,所述突变基因的序列如SEQ ID NO.1所示。
2.一种检测脑海绵状血管瘤的生物标志物,其特征在于,所述生物标志物为权利要求1所述的突变基因。
3.如权利要求1所述的突变基因在制备检测脑海绵状血管瘤试剂、试剂盒或生物芯片中的应用。
4.一种检测脑海绵状血管瘤的试剂盒,其特征在于,所述试剂盒中包括如下引物对:
上游引物:5’-GCCCGGCCAGTAAAATGTTT-3’;
下游引物:5’-TTGAATGAAGCAGGGAACGG-3’。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610503723.2A CN106399470A (zh) | 2016-06-30 | 2016-06-30 | 一种脑海绵状血管瘤突变基因及检测试剂盒 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610503723.2A CN106399470A (zh) | 2016-06-30 | 2016-06-30 | 一种脑海绵状血管瘤突变基因及检测试剂盒 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN106399470A true CN106399470A (zh) | 2017-02-15 |
Family
ID=58006286
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201610503723.2A Withdrawn CN106399470A (zh) | 2016-06-30 | 2016-06-30 | 一种脑海绵状血管瘤突变基因及检测试剂盒 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN106399470A (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114525338A (zh) * | 2022-01-17 | 2022-05-24 | 中山大学 | CCM3、p-Paxillin及p-FAK作为脑海绵状血管瘤的生物标记物的用途 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2001002604A1 (fr) * | 1999-07-01 | 2001-01-11 | Institut National De La Sante Et De La Recherche Medicale (Inserm) | Utilisation du gene krit1 dans le domaine de l'angiogenese |
CN1354185A (zh) * | 2000-11-17 | 2002-06-19 | 上海博德基因开发有限公司 | 一种新的多肽——Krev-1/rapla结合蛋白KRIT134.87和编码这种多肽的多核苷酸 |
-
2016
- 2016-06-30 CN CN201610503723.2A patent/CN106399470A/zh not_active Withdrawn
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2001002604A1 (fr) * | 1999-07-01 | 2001-01-11 | Institut National De La Sante Et De La Recherche Medicale (Inserm) | Utilisation du gene krit1 dans le domaine de l'angiogenese |
CN1354185A (zh) * | 2000-11-17 | 2002-06-19 | 上海博德基因开发有限公司 | 一种新的多肽——Krev-1/rapla结合蛋白KRIT134.87和编码这种多肽的多核苷酸 |
Non-Patent Citations (4)
Title |
---|
DBSNP: "Submitted SNP(ss) Details: ss1705730288", 《DBSNP》 * |
GENBANK: "NCBI Reference Sequence: NM_004912.3", 《GENBANK》 * |
任一昕等: "多发性脑海绵状血管瘤发生的新的CCM1基因突变位点鉴定", 《中国优生与遗传杂志》 * |
伍新田等: "脑海绵状血管瘤分子遗传学研究进展", 《国际神经病学神经外科学杂志》 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114525338A (zh) * | 2022-01-17 | 2022-05-24 | 中山大学 | CCM3、p-Paxillin及p-FAK作为脑海绵状血管瘤的生物标记物的用途 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Koike et al. | Clinicopathological features of acute autonomic and sensory neuropathy | |
Penney Jr et al. | CAG repeat number governs the development rate of pathology in Huntington's disease | |
Desai et al. | A survey of phenotypic features in juvenile polyposis. | |
Carmelli et al. | The Joint Effect of Apolipoprotein E... 4 and MRI Findings on Lower-Extremity Function and Decline in Cognitive Function. | |
Azad et al. | Long-term effectiveness of gross-total resection for symptomatic spinal cord cavernous malformations | |
Mijajlovic et al. | Neurosarcoidosis: Two case reports with multiple cranial nerve involvement and review of the literature | |
CN106399470A (zh) | 一种脑海绵状血管瘤突变基因及检测试剂盒 | |
CN110241208A (zh) | Trem2作为早期诊断冠心病的分子标志物的应用 | |
Klein et al. | Molecular genetic diagnostic program of multiple endocrine neoplasia type 2A and familial medullary thyroid carcinoma syndromes in Hungary | |
WO2023227121A1 (zh) | 一种环状rna在制备脑卒中诊断产品中的应用 | |
Sun et al. | Clinical and genetic analysis of the ABCA4 gene associated retinal dystrophy in a large Chinese cohort | |
Boyadjiev et al. | Linkage analysis narrows the critical region for oculodentodigital dysplasia to chromosome 6q22–q23 | |
Holmberg et al. | Charcot-Marie-Tooth disease in northern Sweden: pedigree analysis and the presence of the duplication in chromosome 17p11. 2. | |
Prasad et al. | Pediatric endocrine screening for von Hippel-Lindau disease: benefits and the challenge of compliance | |
CN104878079A (zh) | 一种用于体外检测Neurofibromastosis2疾病致病基因NF2的c.1598delA突变的试剂盒 | |
CN107247148A (zh) | Gp73的新用途及一种基于其的肝组织炎症活动度检测试剂盒 | |
CN109797218B (zh) | 整合素β4在制备区分结肠癌和直肠癌试剂或药中的应用 | |
Ekström et al. | Detection of alterations in all three exons of the peripherin/RDS gene in Swedish patients with retinitis pigmentosa using an efficient DGGE system. | |
Chan et al. | BamH I polymorphism in the Chinese: its potential usefulness in prenatal diagnosis of beta thalassaemia. | |
Bocquet et al. | Retinitis Punctata Albescens and RLBP1-Allied Phenotypes: Phenotype–Genotype Correlation and Natural History in the Aim of Gene Therapy | |
EP2793028A1 (en) | Composition including thioredoxin 1 as active ingredient, for diagnosis of ovarian cancer or pneumonia, and use thereof | |
Wang et al. | Is FGF23 or FGF21 a promising biomarker to indicate the aging process in COPD? | |
Squitieri et al. | Cavernous angiomas of the nervous system in Italy: clinical and genetic study | |
Ha et al. | Genetic testing identifies the potential risk of multiple endocrine neoplasia in a Vietnamese family | |
Beery et al. | A candidate locus approach identifies a long QT syndrome gene mutation |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
WW01 | Invention patent application withdrawn after publication | ||
WW01 | Invention patent application withdrawn after publication |
Application publication date: 20170215 |