CN103290118B - 一种非缺失型α地中海贫血实时荧光PCR检测试剂盒 - Google Patents
一种非缺失型α地中海贫血实时荧光PCR检测试剂盒 Download PDFInfo
- Publication number
- CN103290118B CN103290118B CN201310191447.7A CN201310191447A CN103290118B CN 103290118 B CN103290118 B CN 103290118B CN 201310191447 A CN201310191447 A CN 201310191447A CN 103290118 B CN103290118 B CN 103290118B
- Authority
- CN
- China
- Prior art keywords
- pcr
- probe
- seq
- pcr reaction
- type
- 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.)
- Active
Links
- 201000006288 alpha thalassemia Diseases 0.000 title claims abstract description 34
- 238000012217 deletion Methods 0.000 title claims abstract description 30
- 230000037430 deletion Effects 0.000 title claims abstract description 30
- 238000001514 detection method Methods 0.000 title abstract description 15
- 238000003752 polymerase chain reaction Methods 0.000 title abstract 7
- 239000000523 sample Substances 0.000 claims abstract description 58
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 16
- 238000012360 testing method Methods 0.000 claims abstract description 15
- KWIUHFFTVRNATP-UHFFFAOYSA-N glycine betaine Chemical compound C[N+](C)(C)CC([O-])=O KWIUHFFTVRNATP-UHFFFAOYSA-N 0.000 claims abstract description 11
- 108090000790 Enzymes Proteins 0.000 claims abstract description 10
- 102000004190 Enzymes Human genes 0.000 claims abstract description 10
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 claims abstract 4
- 229910001629 magnesium chloride Inorganic materials 0.000 claims abstract 2
- 238000006243 chemical reaction Methods 0.000 claims description 42
- 108090000623 proteins and genes Proteins 0.000 claims description 13
- 238000002944 PCR assay Methods 0.000 claims description 10
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 claims description 9
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 claims description 9
- 230000003321 amplification Effects 0.000 claims description 9
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 9
- 238000011144 upstream manufacturing Methods 0.000 claims description 5
- AHCYMLUZIRLXAA-SHYZEUOFSA-N Deoxyuridine 5'-triphosphate Chemical compound O1[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C[C@@H]1N1C(=O)NC(=O)C=C1 AHCYMLUZIRLXAA-SHYZEUOFSA-N 0.000 claims description 4
- SUYVUBYJARFZHO-RRKCRQDMSA-N dATP Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-RRKCRQDMSA-N 0.000 claims description 4
- SUYVUBYJARFZHO-UHFFFAOYSA-N dATP Natural products C1=NC=2C(N)=NC=NC=2N1C1CC(O)C(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-UHFFFAOYSA-N 0.000 claims description 4
- RGWHQCVHVJXOKC-SHYZEUOFSA-J dCTP(4-) Chemical compound O=C1N=C(N)C=CN1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)C1 RGWHQCVHVJXOKC-SHYZEUOFSA-J 0.000 claims description 4
- HAAZLUGHYHWQIW-KVQBGUIXSA-N dGTP Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 HAAZLUGHYHWQIW-KVQBGUIXSA-N 0.000 claims description 4
- 239000011259 mixed solution Substances 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 3
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 3
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 3
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 3
- 125000001309 chloro group Chemical group Cl* 0.000 claims description 3
- 239000012807 PCR reagent Substances 0.000 abstract description 10
- 238000013211 curve analysis Methods 0.000 abstract description 2
- 102000053602 DNA Human genes 0.000 abstract 2
- 108020004414 DNA Proteins 0.000 abstract 2
- 239000002674 ointment Substances 0.000 abstract 2
- 229960003237 betaine Drugs 0.000 abstract 1
- 239000003814 drug Substances 0.000 abstract 1
- 238000009396 hybridization Methods 0.000 abstract 1
- 230000035945 sensitivity Effects 0.000 abstract 1
- 238000012163 sequencing technique Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 26
- 238000000034 method Methods 0.000 description 22
- 238000012408 PCR amplification Methods 0.000 description 13
- 238000010586 diagram Methods 0.000 description 12
- 238000013461 design Methods 0.000 description 8
- 208000002903 Thalassemia Diseases 0.000 description 7
- 108091005902 Hemoglobin subunit alpha Proteins 0.000 description 6
- 238000001712 DNA sequencing Methods 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 238000010791 quenching Methods 0.000 description 4
- 230000000171 quenching effect Effects 0.000 description 4
- 102100027685 Hemoglobin subunit alpha Human genes 0.000 description 3
- 108010054147 Hemoglobins Proteins 0.000 description 3
- 102000001554 Hemoglobins Human genes 0.000 description 3
- 208000005980 beta thalassemia Diseases 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 208000035185 Hemolytic Congenital Anemia Diseases 0.000 description 2
- 206010060893 Hereditary haemolytic anaemia Diseases 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 230000000740 bleeding effect Effects 0.000 description 2
- 201000001516 congenital hemolytic anemia Diseases 0.000 description 2
- -1 dNTPs Proteins 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 208000024987 familial hemolytic anemia Diseases 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 210000005259 peripheral blood Anatomy 0.000 description 2
- 239000011886 peripheral blood Substances 0.000 description 2
- 238000007894 restriction fragment length polymorphism technique Methods 0.000 description 2
- 210000001113 umbilicus Anatomy 0.000 description 2
- 101150084750 1 gene Proteins 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 101150047078 G6PD gene Proteins 0.000 description 1
- 206010064571 Gene mutation Diseases 0.000 description 1
- 206010018910 Haemolysis Diseases 0.000 description 1
- 208000015710 Iron-Deficiency Anemia Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 125000003275 alpha amino acid group Chemical group 0.000 description 1
- 108010053584 alpha-Globins Proteins 0.000 description 1
- 210000001776 amniocyte Anatomy 0.000 description 1
- 210000004381 amniotic fluid Anatomy 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 238000012252 genetic analysis Methods 0.000 description 1
- 108060003196 globin Proteins 0.000 description 1
- 102000018146 globin Human genes 0.000 description 1
- 230000008588 hemolysis Effects 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 238000002205 phenol-chloroform extraction Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- 239000012745 toughening agent Substances 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
Claims (3)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201310191447.7A CN103290118B (zh) | 2013-05-22 | 2013-05-22 | 一种非缺失型α地中海贫血实时荧光PCR检测试剂盒 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201310191447.7A CN103290118B (zh) | 2013-05-22 | 2013-05-22 | 一种非缺失型α地中海贫血实时荧光PCR检测试剂盒 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN103290118A CN103290118A (zh) | 2013-09-11 |
CN103290118B true CN103290118B (zh) | 2014-07-30 |
Family
ID=49091644
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201310191447.7A Active CN103290118B (zh) | 2013-05-22 | 2013-05-22 | 一种非缺失型α地中海贫血实时荧光PCR检测试剂盒 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN103290118B (zh) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104988218A (zh) * | 2015-06-24 | 2015-10-21 | 深圳益生堂生物企业有限公司 | 一种非缺失型α-地中海贫血基因检测的核酸膜条、试剂盒及检测方法 |
CN110257505B (zh) * | 2019-07-02 | 2020-06-05 | 广西壮族自治区人民医院 | 一种非缺失型α地中海贫血点突变快速检测试剂盒及检测方法 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101235415A (zh) * | 2007-01-30 | 2008-08-06 | 中山大学达安基因股份有限公司 | 以TaqMan探针定量多聚酶链反应技术检测基因单点突变的方法 |
CN102277419A (zh) * | 2011-04-06 | 2011-12-14 | 广东省妇幼保健院 | 基于液相芯片系统诊断地中海贫血的方法及试剂盒与应用 |
CN102409100A (zh) * | 2011-11-29 | 2012-04-11 | 潮州市中心医院 | 地中海贫血实时荧光定量pcr检测引物和试剂盒 |
-
2013
- 2013-05-22 CN CN201310191447.7A patent/CN103290118B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101235415A (zh) * | 2007-01-30 | 2008-08-06 | 中山大学达安基因股份有限公司 | 以TaqMan探针定量多聚酶链反应技术检测基因单点突变的方法 |
CN102277419A (zh) * | 2011-04-06 | 2011-12-14 | 广东省妇幼保健院 | 基于液相芯片系统诊断地中海贫血的方法及试剂盒与应用 |
CN102409100A (zh) * | 2011-11-29 | 2012-04-11 | 潮州市中心医院 | 地中海贫血实时荧光定量pcr检测引物和试剂盒 |
Non-Patent Citations (2)
Title |
---|
应用Taqman-MGB探针法检测地中海贫血Hb Westmead突变;陈文强等;《重庆医学》;20121130;第41卷(第32期);第3408页右1-2段 * |
陈文强等.应用Taqman-MGB探针法检测地中海贫血Hb Westmead突变.《重庆医学》.2012,第41卷(第32期),第3408页右1-2段. |
Also Published As
Publication number | Publication date |
---|---|
CN103290118A (zh) | 2013-09-11 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN103602752B (zh) | 一种检测稀有缺失型地中海贫血的引物组和试剂盒 | |
CN102943115B (zh) | α和β地中海贫血基因检测的核酸膜条及试剂盒 | |
CN104099425A (zh) | 一种用于检测B-raf基因突变的试剂盒 | |
CN102876776B (zh) | 检测y染色体微缺失的实时荧光定量pcr试剂盒及方法 | |
CN107119107A (zh) | 一种检测人类cyp2c19基因多态性的方法及试剂盒 | |
CN103436631A (zh) | 一种检测cyp3a5基因多态性的试剂盒及方法 | |
CN104830852B (zh) | 一种检测hla‑b*15:02等位基因的多重实时荧光pcr方法 | |
CN104232781A (zh) | 一种检测HLA-B*5801等位基因的TaqMan探针实时荧光PCR方法 | |
CN102146479A (zh) | 用于精神分裂症、双相情感障碍和重性抑郁症关联基因检测的引物、探针及其试剂盒及制备方法 | |
CN103740855A (zh) | 基于dna条形码的鳗鱼物种鉴别方法 | |
CN106701903A (zh) | 一种用于检测线粒体异质性的试剂盒及检测方法 | |
CN102965427A (zh) | 女性生殖器癌症相关snp位点多重检测方法的建立 | |
CN103290118B (zh) | 一种非缺失型α地中海贫血实时荧光PCR检测试剂盒 | |
CN102925560A (zh) | HRM法检测突变型α-地中海贫血基因的试剂盒和方法 | |
CN108070636A (zh) | 一种荧光pcr扩增样本的处理方法和试剂盒 | |
CN105018490A (zh) | 用于检测人类mthfr基因多态性的引物对、探针及试剂盒 | |
CN108949929A (zh) | 用于同时检测mthfr和mtrr基因多态性的产品及其方法和应用 | |
CN103205485B (zh) | 一种检测蜡样芽孢杆菌核苷酸的方法及检测用引物和探针 | |
CN101985659A (zh) | 检测精神分裂关联基因的试剂盒及其制备方法 | |
CN107022620A (zh) | N‑ras基因突变检测引物探针及其试剂盒 | |
CN103993089A (zh) | 一种耐万古霉素肠球菌的多重荧光定量pcr检测试剂盒及其检测方法 | |
CN104293968B (zh) | 一组用于检测非综合征性唇腭裂遗传易感性的多态性位点及检测试剂盒 | |
CN106755422B (zh) | 一种与黄牛生长性状相关的meg3基因snp的检测方法及其应用 | |
CN105586392A (zh) | 评估胎儿样本中母体细胞污染程度的方法 | |
CN115029460A (zh) | 一种副猪嗜血杆菌的即时可视化检测方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
TR01 | Transfer of patent right |
Effective date of registration: 20240226 Address after: 518000, Room 02, 43rd Floor, Building D1, Kexing Science Park, No. 15 Keyuan Road, Science Park Community, Yuehai Street, Nanshan District, Shenzhen, Guangdong Province Patentee after: CHUANGYI BIOTECHNOLOGY Co.,Ltd. Country or region after: China Address before: 518000, 2nd floor, Building 6, Nanyou Yuehai Industrial Zone, Nanshan District, Shenzhen, Guangdong Province Patentee before: YISHENGTANG BIOLOG ENTPR Co.,Ltd. Country or region before: China Patentee before: CHUANGYI BIOTECHNOLOGY Co.,Ltd. |
|
TR01 | Transfer of patent right |