CN102586433A - Deafness predisposing gene 12S rRNA (ribosomal ribonucleic acid) 1494C>T fluorescence detection kit and application thereof - Google Patents
Deafness predisposing gene 12S rRNA (ribosomal ribonucleic acid) 1494C>T fluorescence detection kit and application thereof Download PDFInfo
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Abstract
The invention discloses a fluorescence detection kit for detecting deafness predisposing gene 12S rRNA (ribosomal ribonucleic acid) 1494C>T. The kit comprises a reagent before amplification and a reagent after amplification, wherein the reagent before amplification comprises a reaction mixture of a PCR (polymerase chain reaction) buffer solution, MgCl2 and dNTPs (deoxyribonucleoside triphosphate), Taq enzyme, ultrapure water, and a primer mixture for high-specific amplification of 12SrRNA1494C>T and amelogenin locus; and the reagent after amplification comprises an allelic ladder and an internal standard. According to the invention, 12SrRNA1494C>T and amelogenin locus are used as detecting objects, so that the fluorescence detection kit has important meaning to detection of deafness predisposing genes, particularly screening of newborn deafness genes.
Description
Technical field
The present invention relates to the fluorescence detection reagent kit of a kind of deaf sick tumor susceptibility gene 12S rRNA 1494C＞T of detection and Amelogenin locus, belong to technical field of biological.
Background technology
The paathogenic factor research of in the world wide hearing and speech handicapped person being carried out shows that about 60-80% patient's the cause of disease is relevant with inherited genetic factors, and wherein the clinical study data of developed country show that hereditary hearing impairment accounts for 80% in deafness patient.Therefore in recent ten years, the pathogenesis of hereditary hearing impairment and the research of molecular epidemiology thereof become one of deaf sick research important contents.Along with the Human Genome Project is accomplished; The location and the clone of deaf ospc gene have obtained great advance, the molecule genetics research of deaf disease and the data of molecular epidemiology make investigators progressively recognize deaf sick susceptibility gene mutation safeguard hearing healthy with find the importance of hearing in unusual.
The ototoxicity of medicine is an important factor of hearing loss before causing speaking; Part is relevant with plastosome 12S rRNA transgenation; In Chinese colony, found the relation of 12S rRNA gene 1494C＞T sudden change, had been found that three big familys are owing to 1494C＞T sudden change causes at present at least with drug induced deafness.Often hearing is normal at birth for 1494C＞T sudden change person, is difficult to found in advance through the hearing examination and predict, but has the hidden danger that hearing loss takes place because of the use of ototoxic drug.
Gene tester commonly used at present has: (direct sequencing directly checks order; DS), ligase enzyme detection reaction (ligase detection reaction; LDR), restriction fragment length polymorphism analysis (restriction fragment length polymorphism; RFLP), dhplc analysis (denaturing high performance liquid chromotography, DHPLC), quantitative PCR, gene chip.All there is different defects in these methods, and for example complex operation, result are difficult for problems such as interpretation, poor repeatability, false negative false positive be many.
Adopt fluorescent mark technology, capillary electrophoresis technique; Through the band fluorescent marker primer to the site-specific amplification of 12S rRNA 1494C＞T; And the Tm value of in this primer, mixing nucleic acid analog raising primer; Further strengthen the specificity of primer, and, can realize detection the deaf gene site after capillary electrophoresis post analysis PCR product goes out the peak situation.This method is highly sensitive, interpretation is clear accurately only need take a small amount of blood sample or buccal swab sample, can detect the convenient collection that especially is fit to the newborn infants sample of sampling.
Summary of the invention
The object of the invention provides a kind of deaf sick tumor susceptibility gene 12S rRNA 1494C＞T fluorescence detection reagent kit.
To achieve these goals, the technical scheme of taking: the gene type after a kind of deaf sick tumor susceptibility gene 12S rRNA 1494C＞T fluorescence detection reagent kit, this test kit comprise amplifing reagent and increase is used reagent;
Reagent comprises before the said amplification: PCR damping fluid, MgCl
2, dNTPs mixture, Taq enzyme, the 2 pairs of primer mixtures and ultrapure water;
Said amplification back reagent comprises: be used for detecting and the mixture and the interior mark ROX-300 of the allelotrope standard substance of Amelogenin sex identification locus corresponding to 12S rRNA 1494C＞T of gene type;
Use described test kit to carry out the condition of PCR composite amplification reaction: the pH value of pcr amplification system is 8.0-9.0; Magnesium ion concentration is 1.5-3.5mM; The final concentration of 4 kinds of dNTP respectively is 200-300 μ M; The consumption of Taq enzyme is 0.1-0.4U/ μ L, and the list in the 2 pairs of primer mixtures is 0.2-0.4 μ M to the primer final concentration.
When using said test kit to carry out pcr amplification, amplification elementary reaction in a composite amplification reaction system, increase simultaneously 12S rRNA 1494C＞T and Amelogenin locus.
Being used for the primer that 12S rRNA 1494C＞the T sudden change detects is: SEQ NO.01 and SEQ NO.02.
SEQ NO.02 is the primer (the LNA nucleosides is represented with italics) that has mixed the LNA nucleosides:
SEQ?NO.1：5’-AGTGGGTTTGGGGCTAGGTTTA-3’
SEQ?NO.2：5’-GACACACCGCCCGTCTCT-3’
The primer that is used for the detection of Amelogenin locus is: SEQ NO.03:5 '-CCCTGGGCTCTGTAAAGAAT-3 ', SEQ NO.04:5 '-GCAGAGCTTAAACTGGGAAGC-3 '.
Described 2 pairs of primers wherein have 5 ' end of a primer to carry out fluorochrome label in every pair of primer, and used optical dye can be identical or different resorcinolphthalein.
Said composite amplification adopts the polymerase chain reaction to realize, adopts multiple tracks or single track capillary electrophoresis to detect.
The human gene group DNA who is wherein detected is: use Chelex method, magnetic bead extraction method or phenol/chloroform extraction method that the source sample is handled the DNA that obtains; Described source sample is: derive from human: filter paper blood cake/buccal swab sample, FTA card blood cake/buccal swab sample, buccal swab sample, blood/trace, tissue, amniotic fluid.
When using described test kit to carry out pcr amplification, the amplification elementary reaction carries out amplification program on the PCR of any model appearance: 94-98 ℃ of 1-5min; 26-32 round-robin 94-98 ℃ 15-60s, 55-65 ℃ of 15-60s, 72 ℃ of 15-60; 60 ℃ of 30-60min.
For detecting of 12S rRNA 1494C＞T sudden change; The band fluorescent mark that uses also mixes the primer of modification through the LNA nucleoside monomers, has improved the Tm value of primer, has shortened primer length; Thereby increased the sensitivity and the specificity of primer, prevented false positive and false-negative appearance.
For detecting of Amelogenin, be the effect of confidential reference items on the one hand: whether the indication template DNA is effectively or concentration range; Can effectively indicate whether to exist the PCR product pollution on the other hand, prevent the false positive that produces because of polluting.
Description of drawings
Fig. 1 proves conclusively the somatotype result of case different concns DNA through the primer of modifying to the same male sex.
Fig. 2 is that the primer of unmodified is proved conclusively the somatotype result of case different concns DNA to the same male sex.
Fig. 3 is through the primer the modified somatotype result to same male sex's normal individual different concns DNA.
Fig. 4 is the somatotype result of the primer of unmodified to same male sex's normal individual different concns DNA.
Embodiment
Below in conjunction with accompanying drawing and specific embodiment the present invention is described further, but not as to qualification of the present invention.
Embodiment 1
Deaf sick tumor susceptibility gene 12S rRNA 1494C＞T fluorescence detection reagent kit detects the dna sample of sudden change and normal individual, is used for the primer that deaf sick tumor susceptibility gene 12S rRNA 1494C＞T detects, the primer that is used for the amplification of Amelogenin locus all adopts Yellow fluorochrome TAMRA mark; Interior mark adopts the red fluorescence dyestuff mark, and fluorescent marker is ROX.Wherein, be used for the primer use that deaf sick tumor susceptibility gene 12S rRNA 1494C＞T detects: the primer (SEQ NO.2:5 '-GACACACCGCCCGTCTCT-3 ') and the general primer (SEQ NO.2.0:5 '-GACACACCGCCCGTCTCT-3 ') that mix modification through the LNA nucleoside monomers carry out the comparison of expanding effect.
1, sample to be tested is 100 parts, and the technological method that all uses " DNA extraction-pcr amplification-order-checking " carried out order-checking to 12S rRNA 1494 sites and detects, and 2 parts in sample wherein suddenlys change.
2, the extracting genome DNA of sample
Chelex extraction method: cut 1～3mm blood cake and place the 1.5mL centrifuge tube, add sdH
2O 1mL, it is centrifugal to vibrate, and abandons supernatant, and repeating step twice is abandoned supernatant, and draw 200 μ L with the rifle head of cutting fast after 5% Chelex-100 concussion is suspended and add in the centrifuge tube, the vibration several seconds.Behind 56 ℃ of water bath heat preservation 30min, the vibration several seconds.95 ℃ of boiling water bath 10min slightly vibrate the several seconds.The centrifugal 5min of 2000rpm, the DNA for extracting in the supernatant.
3, the check and analysis of amplification and amplified production
3.1 pcr amplification system:
3.2 pcr amplification program:
4, amplified production fluoroscopic examination on genetic analyzer
Form appearance mixture ((mark in the 0.2 μ L molecular weight) * (sample introduction number)+(9.8 μ L deionized formamide) * (sample introduction number)) by mark in deionized formamide and the molecular weight.Appearance mixture on the 10 μ L is mixed with 1 μ L amplified production or allelotrope analytical standard, avoid producing bubble.95 ℃ of sex change 5min, ice bath 5min, and electrophoresis as early as possible.Use the genetic analyzer check and analysis.
5, conclusion
Like Fig. 1, shown in 2, for male sex's confirmed cases,
Primer SEQ NO.2 (test kit primer of the present invention) the gained result (Fig. 1) who has mixed the LNA nucleoside monomers is with consistent without the SEQ NO.2.0 primer gained result (Fig. 2) who modifies; Promptly under each DNA concentration, all can obtain correct result of determination: 12S rRNA 1494C＞interpretation position, T site goes out the peak; Sex goes out X, Y is bimodal; All the increase peak value with DNA concentration increases, and is specific as follows:
Like Fig. 3, shown in 4, for male sex's normal individual,
Mixed primer SEQ NO.2 (test kit primer of the present invention) the gained result (Fig. 3) of LNA nucleoside monomers and inconsistent to 12S rRNA 1494C＞T site without the SEQ NO.2.0 primer gained result (Fig. 4) who modifies: the former does not all go out the peak under each DNA concentration; And the latter is when the amount of DNA during greater than 2ng, and 1494C＞interpretation position, T site goes out the peak, and is inconsistent with sequencing result, promptly uses without the SEQ NO.2.0 primer of modifying and is prone to false-positive result.Both are consistent for the judgement of sex: sex goes out X, Y is bimodal, and all the increase peak value with DNA concentration increases.Specific as follows:
Therefore, test kit primer of the present invention can detect 12S rRNA 1494C＞T site mutation specifically, and the result is consistent with sequencing result, false positive do not occur.
2.5 * PCR damping fluid of used different pH among the above embodiment, with the Tris-HCl damping fluid preparation of different pH values, the Tris-HCl concentration in 1 * PCR damping fluid is 10mM, KCl concentration is 50mM; Used Taq polysaccharase and other reagent and material are the commercially available prod among the present invention.
Claims (7)
1. deaf sick tumor susceptibility gene 12S rRNA 1494C＞T fluorescence detection reagent kit is characterized in that comprising the gene type standard substance and the interior mark of primer, gene mutation site 12S rRNA 1494C＞T and the Amelogenin locus of archaeal dna polymerase and buffered soln, amplification 12S rRNA 1494C＞T and Amelogenin locus.
2. test kit according to claim 1 is characterized in that the primer that mutantional hotspot 12S rRNA 1494C＞T detects is: SEQ ID NO.1 and SEQ ID NO.2;
SEQ?NO.1：5’-AGTGGGTTTGGGGCTAGGTTTA-3’
SEQ?NO.2：5’-GACACACCGCCCGTCTCT-3’
Wherein the LNA nucleosides is represented with italics.
3. test kit according to claim 1 and 2 is characterized in that the primer that said Amelogenin locus detects is:
SEQ?NO.3：5’-CCCTGGGCTCTGTAAAGAAT-3’
SEQ?NO.4：5’-GCAGAGCTTAAACTGGGAAGC-3’。
4. test kit according to claim 1 and 2 is characterized in that: the detection primer of said 12S rRNA 1494C＞T and Amelogenin has 5 ' end of a primer to carry out fluorochrome label in every pair of primer.
5. test kit according to claim 1 is characterized in that: the primer that said 12S rRNA 1494C＞T detects can adopt identical or different fluorochrome label with the primer that is used for the amplification of Amelogenin locus; Interior mark is selected for use and is different from above-mentioned fluorochrome label.
6. test kit according to claim 1 is characterized in that: the composite amplification of said 12S rRNA 1494C＞T and Amelogenin locus adopts the polymerase chain reaction to realize, adopts multiple tracks or single track capillary electrophoresis to detect.
7. application rights requires 1 said test kit to detect the method for deaf sick tumor susceptibility gene 12S rRNA 1494C＞T, it is characterized in that through fluorescent primer PCR and deaf sick tumor susceptibility gene 12S rRNA 1494C＞T of capillary electrophoresis specific detection and Amelogenin locus; The primer that is used for 12S rRNA 1494C＞T sudden change detection is shown in SEQ NO.1 and SEQ NO.2; The primer that is used for the detection of Amelogenin locus is shown in SEQ NO.3, SEQ NO.4; The composite amplification of said 12S rRNA 1494C＞T and Amelogenin locus adopts the polymerase chain reaction to realize, adopts multiple tracks or single track capillary electrophoresis to detect.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102796822A (en) * | 2012-08-30 | 2012-11-28 | 山东百福基因科技有限公司 | Motion function gene ACTN3 (alpha-Actinin) fluorescent detection kit and detection method |
| CN102796823A (en) * | 2012-08-30 | 2012-11-28 | 山东百福基因科技有限公司 | Fluorescence detection kit and detection method for motor function related gene CKMM |
| CN103451302A (en) * | 2013-09-11 | 2013-12-18 | 步迅 | Deafness susceptible gene mitochondrion 12SrDNA 1555A>G and 1494C>T mutant ratio detection kit |
| CN104419757A (en) * | 2013-09-04 | 2015-03-18 | 中南大学湘雅医院 | Novel deafness related gene mutation detection system and kit |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1332805A (en) * | 1998-11-25 | 2002-01-23 | 普罗梅加公司 | Multiplex amplification of short tandem repeat loci |
| CN1361829A (en) * | 1999-03-18 | 2002-07-31 | 埃克西库恩公司 | Detection of mutations in genes by specific LNA primers |
| CN1733917A (en) * | 2004-09-17 | 2006-02-15 | 中国人民解放军总医院 | Human phonosensitive nerve deafness related WFS1 mutant gene and its uses in diagnosing genetic deafness |
| CN101597638A (en) * | 2008-06-04 | 2009-12-09 | 博奥生物有限公司 | A kind of method and dedicated kit thereof that detects gene multi-mutant site |
| CN101684497A (en) * | 2008-09-23 | 2010-03-31 | 中国人民解放军总医院 | Deafness susceptibility gene screen test kit |
| CN102304584A (en) * | 2011-09-15 | 2012-01-04 | 上海交通大学医学院附属新华医院 | GJB2 gene p.V37I mutation diagnostic kit for diagnosing late-onset deafness |
-
2012
- 2012-02-14 CN CN201210033106.2A patent/CN102586433B/en not_active Expired - Fee Related
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1332805A (en) * | 1998-11-25 | 2002-01-23 | 普罗梅加公司 | Multiplex amplification of short tandem repeat loci |
| CN1361829A (en) * | 1999-03-18 | 2002-07-31 | 埃克西库恩公司 | Detection of mutations in genes by specific LNA primers |
| CN1733917A (en) * | 2004-09-17 | 2006-02-15 | 中国人民解放军总医院 | Human phonosensitive nerve deafness related WFS1 mutant gene and its uses in diagnosing genetic deafness |
| CN101597638A (en) * | 2008-06-04 | 2009-12-09 | 博奥生物有限公司 | A kind of method and dedicated kit thereof that detects gene multi-mutant site |
| CN101684497A (en) * | 2008-09-23 | 2010-03-31 | 中国人民解放军总医院 | Deafness susceptibility gene screen test kit |
| CN102304584A (en) * | 2011-09-15 | 2012-01-04 | 上海交通大学医学院附属新华医院 | GJB2 gene p.V37I mutation diagnostic kit for diagnosing late-onset deafness |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102796822A (en) * | 2012-08-30 | 2012-11-28 | 山东百福基因科技有限公司 | Motion function gene ACTN3 (alpha-Actinin) fluorescent detection kit and detection method |
| CN102796823A (en) * | 2012-08-30 | 2012-11-28 | 山东百福基因科技有限公司 | Fluorescence detection kit and detection method for motor function related gene CKMM |
| CN104419757A (en) * | 2013-09-04 | 2015-03-18 | 中南大学湘雅医院 | Novel deafness related gene mutation detection system and kit |
| CN103451302A (en) * | 2013-09-11 | 2013-12-18 | 步迅 | Deafness susceptible gene mitochondrion 12SrDNA 1555A>G and 1494C>T mutant ratio detection kit |
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| CN102586433B (en) | 2014-01-29 |
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