CN104099402A - 单管荧光PCR技术检测GJB2基因235delC突变的方法及其试剂盒 - Google Patents
单管荧光PCR技术检测GJB2基因235delC突变的方法及其试剂盒 Download PDFInfo
- Publication number
- CN104099402A CN104099402A CN201310116474.8A CN201310116474A CN104099402A CN 104099402 A CN104099402 A CN 104099402A CN 201310116474 A CN201310116474 A CN 201310116474A CN 104099402 A CN104099402 A CN 104099402A
- Authority
- CN
- China
- Prior art keywords
- wild
- probe
- type probe
- gjb2 gene
- gjb2
- 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.)
- Pending
Links
- 101150034593 Gjb2 gene Proteins 0.000 title claims abstract description 35
- 238000000034 method Methods 0.000 title claims abstract description 31
- 230000035772 mutation Effects 0.000 title claims abstract description 15
- 238000005516 engineering process Methods 0.000 title abstract description 3
- 239000000523 sample Substances 0.000 claims abstract description 96
- 238000006243 chemical reaction Methods 0.000 claims abstract description 26
- 230000003321 amplification Effects 0.000 claims abstract description 16
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 16
- 239000003086 colorant Substances 0.000 claims abstract description 7
- 238000003908 quality control method Methods 0.000 claims description 35
- 238000012360 testing method Methods 0.000 claims description 24
- 101000954092 Homo sapiens Gap junction beta-2 protein Proteins 0.000 claims description 16
- 102100037156 Gap junction beta-2 protein Human genes 0.000 claims description 15
- 238000013461 design Methods 0.000 claims description 14
- 239000000243 solution Substances 0.000 claims description 14
- 239000002773 nucleotide Substances 0.000 claims description 10
- 125000003729 nucleotide group Chemical group 0.000 claims description 10
- 239000007850 fluorescent dye Substances 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 210000004369 blood Anatomy 0.000 claims description 8
- 239000008280 blood Substances 0.000 claims description 8
- 108090000790 Enzymes Proteins 0.000 claims description 6
- 102000004190 Enzymes Human genes 0.000 claims description 6
- 238000003556 assay Methods 0.000 claims description 6
- 230000000869 mutational effect Effects 0.000 claims description 6
- 239000013600 plasmid vector Substances 0.000 claims description 6
- 241000972773 Aulopiformes Species 0.000 claims description 4
- 238000012408 PCR amplification Methods 0.000 claims description 4
- 239000012634 fragment Substances 0.000 claims description 4
- 238000010791 quenching Methods 0.000 claims description 4
- 230000000171 quenching effect Effects 0.000 claims description 4
- 235000019515 salmon Nutrition 0.000 claims description 4
- -1 dNTP Substances 0.000 claims description 3
- 239000008367 deionised water Substances 0.000 claims description 3
- 229910021641 deionized water Inorganic materials 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 238000000605 extraction Methods 0.000 claims description 3
- 108020004707 nucleic acids Proteins 0.000 claims description 3
- 150000007523 nucleic acids Chemical class 0.000 claims description 3
- 102000039446 nucleic acids Human genes 0.000 claims description 3
- 239000002504 physiological saline solution Substances 0.000 claims description 3
- 210000001776 amniocyte Anatomy 0.000 claims description 2
- 238000012217 deletion Methods 0.000 claims description 2
- 230000037430 deletion Effects 0.000 claims description 2
- 230000008034 disappearance Effects 0.000 claims description 2
- 238000003752 polymerase chain reaction Methods 0.000 claims description 2
- 210000003296 saliva Anatomy 0.000 claims description 2
- 238000011144 upstream manufacturing Methods 0.000 claims description 2
- 239000000463 material Substances 0.000 claims 2
- 206010011878 Deafness Diseases 0.000 abstract description 11
- 231100000895 deafness Toxicity 0.000 abstract description 7
- 208000016354 hearing loss disease Diseases 0.000 abstract description 7
- 238000003745 diagnosis Methods 0.000 abstract description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 4
- 238000004458 analytical method Methods 0.000 abstract description 3
- 201000010099 disease Diseases 0.000 abstract description 3
- 239000000969 carrier Substances 0.000 abstract description 2
- 206010011882 Deafness congenital Diseases 0.000 abstract 1
- 239000013062 quality control Sample Substances 0.000 abstract 1
- 238000012216 screening Methods 0.000 abstract 1
- 239000000047 product Substances 0.000 description 31
- 238000001514 detection method Methods 0.000 description 10
- 239000007788 liquid Substances 0.000 description 9
- 238000007400 DNA extraction Methods 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 5
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 210000003743 erythrocyte Anatomy 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000002093 peripheral effect Effects 0.000 description 4
- 229910001425 magnesium ion Inorganic materials 0.000 description 3
- 230000007018 DNA scission Effects 0.000 description 2
- 238000011529 RT qPCR Methods 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 238000013016 damping Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 238000001502 gel electrophoresis Methods 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 238000012856 packing Methods 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- 238000007894 restriction fragment length polymorphism technique Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108010014303 DNA-directed DNA polymerase 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
- 206010064571 Gene mutation Diseases 0.000 description 1
- 238000002944 PCR assay Methods 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 238000007844 allele-specific PCR Methods 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- 208000021024 autosomal recessive inheritance Diseases 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 206010025482 malaise Diseases 0.000 description 1
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Natural products C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 1
- 201000006790 nonsyndromic deafness Diseases 0.000 description 1
- 230000035935 pregnancy Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000009863 secondary prevention Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 230000001360 synchronised effect Effects 0.000 description 1
- ABZLKHKQJHEPAX-UHFFFAOYSA-N tetramethylrhodamine Chemical compound C=12C=CC(N(C)C)=CC2=[O+]C2=CC(N(C)C)=CC=C2C=1C1=CC=CC=C1C([O-])=O ABZLKHKQJHEPAX-UHFFFAOYSA-N 0.000 description 1
- 238000012073 universal newborn hearing screening Methods 0.000 description 1
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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6851—Quantitative amplification
-
- 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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6858—Allele-specific amplification
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
Claims (7)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201310116474.8A CN104099402A (zh) | 2013-04-07 | 2013-04-07 | 单管荧光PCR技术检测GJB2基因235delC突变的方法及其试剂盒 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201310116474.8A CN104099402A (zh) | 2013-04-07 | 2013-04-07 | 单管荧光PCR技术检测GJB2基因235delC突变的方法及其试剂盒 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN104099402A true CN104099402A (zh) | 2014-10-15 |
Family
ID=51667940
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201310116474.8A Pending CN104099402A (zh) | 2013-04-07 | 2013-04-07 | 单管荧光PCR技术检测GJB2基因235delC突变的方法及其试剂盒 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN104099402A (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111321218A (zh) * | 2020-04-14 | 2020-06-23 | 上海交通大学医学院附属第九人民医院 | 迟发性耳聋GJB2基因pV37I突变的检测试剂盒 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101684493A (zh) * | 2008-09-26 | 2010-03-31 | 上海裕隆生物科技有限公司 | 一种儿童先天性耳聋基因分析检测试剂盒 |
CN101684497A (zh) * | 2008-09-23 | 2010-03-31 | 中国人民解放军总医院 | 一种耳聋易感基因筛查试剂盒 |
CN102465174A (zh) * | 2010-11-09 | 2012-05-23 | 王宝恒 | 荧光定量pcr技术检测gjb2基因突变的方法及其试剂盒 |
CN102618624A (zh) * | 2011-01-26 | 2012-08-01 | 苏州市立医院 | 中国人群耳聋基因筛查用试剂盒及其使用方法 |
-
2013
- 2013-04-07 CN CN201310116474.8A patent/CN104099402A/zh active Pending
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101684497A (zh) * | 2008-09-23 | 2010-03-31 | 中国人民解放军总医院 | 一种耳聋易感基因筛查试剂盒 |
CN101684493A (zh) * | 2008-09-26 | 2010-03-31 | 上海裕隆生物科技有限公司 | 一种儿童先天性耳聋基因分析检测试剂盒 |
CN102465174A (zh) * | 2010-11-09 | 2012-05-23 | 王宝恒 | 荧光定量pcr技术检测gjb2基因突变的方法及其试剂盒 |
CN102618624A (zh) * | 2011-01-26 | 2012-08-01 | 苏州市立医院 | 中国人群耳聋基因筛查用试剂盒及其使用方法 |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111321218A (zh) * | 2020-04-14 | 2020-06-23 | 上海交通大学医学院附属第九人民医院 | 迟发性耳聋GJB2基因pV37I突变的检测试剂盒 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN1582341A (zh) | 分离细胞的方法及其使用 | |
CN107058538B (zh) | 一种引物组合物及其组成的试剂盒和应用 | |
GB2488358A (en) | Enrichment of foetal DNA in maternal plasma | |
CN107254531B (zh) | 早发性结直肠癌辅助诊断的遗传生物标志物及其应用 | |
CN102465174A (zh) | 荧光定量pcr技术检测gjb2基因突变的方法及其试剂盒 | |
WO2014139330A1 (en) | Rapid genotyping analysis and kits thereof | |
CN105861641A (zh) | 一种用于检测cho细胞dna残留的引物、试剂盒及检测方法 | |
EP2964780B1 (en) | Discrimination of blood type variants | |
CN111118138A (zh) | 一种叶酸代谢能力基因mthfr和mtrr多态性检测试剂盒及方法 | |
CN106521023A (zh) | 一种检测中国人HPFH和δβ‑地中海贫血的试剂盒及方法 | |
CN101555528B (zh) | 一种染色体22q11.2区微缺失、微重复的测定方法 | |
US20150024390A1 (en) | Method and kit for detecting specific single nucleotide polymorphism associated with ankylosing spondylitis | |
Ramezanzadeh et al. | Detection of paternally inherited fetal point mutations for β-thalassemia in maternal plasma using simple fetal DNA enrichment protocol with or without whole genome amplification: an accuracy assessment | |
CN108715893B (zh) | 一组与放疗引起的放射性脑损伤相关的snp标志物及其应用 | |
CN104120166A (zh) | 荧光pcr技术检测pds基因2168 a> g突变的方法及其试剂盒 | |
CN109182493A (zh) | 人16p11.2微缺失综合征检测的引物和试剂盒及其检测方法 | |
CN104087672A (zh) | 一种多重实时荧光定量pcr技术快速检测人21号染色体数目的试剂盒 | |
Meaney et al. | Noninvasive prenatal diagnosis of early onset primary dystonia I in maternal plasma | |
CN103502469A (zh) | 强直性脊柱炎易感性单核苷酸多态性的检测方法、试剂盒及其应用 | |
CN104087671A (zh) | 一种用于检测人21号染色体数目的试剂盒 | |
CN104099402A (zh) | 单管荧光PCR技术检测GJB2基因235delC突变的方法及其试剂盒 | |
EP2450455A1 (en) | Method for determining presence or absence of epithelial cancer-origin cell in biological sample, and molecular marker and kit therefor | |
CN106566880A (zh) | 遗传性耳聋基因突变检测试剂盒 | |
CN108504731B (zh) | 诊断脂质代谢相关疾病或阿尔茨海默病标志物的方法 | |
CN104120167A (zh) | 荧光pcr技术检测pds基因ivs7-2a>g突变的方法及其试剂盒 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
ASS | Succession or assignment of patent right |
Owner name: SHI GUIZHI Free format text: FORMER OWNER: WANG BAOHENG Effective date: 20150213 Owner name: WANG BAOHENG Effective date: 20150213 |
|
C41 | Transfer of patent application or patent right or utility model | ||
C53 | Correction of patent of invention or patent application | ||
CB03 | Change of inventor or designer information |
Inventor after: Shi Guizhi Inventor after: Wang Baoheng Inventor before: Wang Baoheng Inventor before: Shi Guizhi |
|
COR | Change of bibliographic data |
Free format text: CORRECT: INVENTOR; FROM: WANG BAOHENG SHI GUIZHI TO: SHI GUIZHI WANG BAOHENG |
|
TA01 | Transfer of patent application right |
Effective date of registration: 20150213 Address after: 250002, room 2, building 9, building 501, provincial court dormitory, sunshine Shun two district, Shizhong District, Ji'nan City, Shandong Applicant after: Shi Guizhi Applicant after: Wang Baoheng Address before: 250002, room 2, building 9, building 501, provincial court dormitory, sunshine Shun two district, Shizhong District, Ji'nan City, Shandong Applicant before: Wang Baoheng |
|
DD01 | Delivery of document by public notice |
Addressee: Shi Guizhi Document name: Notification to Make Formalities Rectification |
|
WD01 | Invention patent application deemed withdrawn after publication | ||
WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20141015 |