CN103602672A - Primer, probe and kit for detection of gene expression amounts and use methods thereof - Google Patents
Primer, probe and kit for detection of gene expression amounts and use methods thereof Download PDFInfo
- Publication number
- CN103602672A CN103602672A CN201310537253.8A CN201310537253A CN103602672A CN 103602672 A CN103602672 A CN 103602672A CN 201310537253 A CN201310537253 A CN 201310537253A CN 103602672 A CN103602672 A CN 103602672A
- Authority
- CN
- China
- Prior art keywords
- seq
- probe
- primer
- gene
- sequence
- 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.)
- Granted
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a primer, a probe and a kit for detection of gene expression amounts and uses method thereof, and the invention also discloses the kit comprising the primer and the probe and a use method of the kit. The sequences of the primer are shown as SEQ ID NO:1 to SEQ ID NO:8, the sequences of the probe are shown as SEQ ID NO:11 to SEQ IDNO:14 shows, the kit includes the primer, the probe, a primer and a probe of a reference gene, a real-time fluorescent quantitative PCR (polymerase chain reaction) MIX reaction solution, deionized water, bottles or tubes for separated and concentrated packaging of the reagents, and a packing box. The primer, the probe and the kit can detect the expression amounts of four genes, the detection range is wide, the detection process is simple and quick, and the primer, the probe and the kit help a doctor to conduct a comprehensive analysis and judgment of a medication strategy of a patient.
Description
Technical field
The invention belongs to technical field of molecular biology, relate to primer, probe and the test kit of gene test, relate in particular to primer, probe and test kit, using method and the application of a kind of ERCC1 of detection, RRM1, TUBB3, TYMS gene expression amount.
Background technology
Along with medical research and medicament research and development level constantly improve, increasing treatment tumour class medicine is developed, but, some patients effect when using some drugs to treat is unsatisfactory, show as tumour low to Susceptibility, resistance is strong etc., and research shows that this phenomenon is relevant with some gene overexpression in human body.
Platinum medicine is one of the most frequently used tumor chemotherapeutic drug, and it is mainly by causing the adducts that is cross-linked to form of DNA in tumour cell, hinder DNA synthetic with copy, thereby the growth of inhibition tumor cell.ERCC1 is nucleic acid excision repair system NER(nucleotide excision repair) a key gene, NER can excise adducts and repair the DNA being damaged, thereby reduction chemotherapy effect, therefore, the high expression level of ERCC1 in mRNA is relevant with the resistance of platinum medicine.
Gemcitabine is a kind of antitumor drug that is used for clinically treating nonsmall-cell lung cancer, mammary cancer, carcinoma of the pancreas and other noumenal tumours, medicine enters after cell, can activate by a series of phosphokinase, generate nucleoside triphosphate analogue, thereby the substrate competition synthetic with DNA mixes DNA chain, interrupt the synthetic of DNA.Clinical study shows, the expression level of RRM1 gene and gemcitabine curative effect negative correlation, the Patients with Non-small-cell Lung that RRM1mRNA expression level is low, after accepting gemcitabine and platinum medicine combined chemotherapy, have preferably and reply, its meta Overall survival is also long than the high patient of RRM1mRNA expression level, in addition, the high expression level of RRM1 in carcinoma of the pancreas also can cause patient to produce resistance to gemcitabine, the comprehensive cancer network of American National nonsmall-cell lung cancer clinical practice guideline (2011) is the prediction biomarker using RRM1 expression level as treatment, therefore, detecting RRM1 expression level is very important for the curative effect of predicting gemcitabine, it is that main chemotherapeutic efficacy is better that the patient that RRM1 expression level is low adopts gemcitabine.
Anti-microtubule based chemotherapy medicine is to act on cell microtubule, by affecting spindle body, form, a thereby class spectrum chemotherapeutic that suppresses cell mitogen, intracellular canaliculus albumen is divided into α, two hypotypes of β, it is the Chinese medicine integral part of cytoskeleton, the basic composition unit of spindle body while being mitotic division is also the Main Function target spot of breast cancer.Show according to the study, β-Tubulin-III of TUBB3 genes encoding (3 type beta tubulin) is the closest with the relation of anti-microtubule chemosensitivity perception, it is better that the tumour patient of low TUBB3mRNA expression level is accepted the effect of anti-microtubule class medicine chemotherapy, median survival interval is longer, otherwise the anti-microtubule class medicine chemotherapeutic efficacy of the tumour patient of high TUBB3mRNA expression level is poor.
Fluorine class medicine is the fluoro derivatives of uridylic, can in cell, change 5 FU 5 fluorouracil deoxynucleotide (5F-dUMP) into, thereby suppress deoxynucleoside acid enzyme, stop deoxynucleotide (dUMP) methyl to turn to deoxythymidylic acid (dTMP), and then affect the synthetic of DNA, in addition, also can mix the synthetic of interferencing protein in RNA, thereby reach the effect that suppresses tumor growth.TYMS(Thymidylate Synthetase) thymidylate synthetase of genes encoding (TS) is the synthetic rate-limiting enzyme of pyrimidine nucleotide, it is the important factor of tumor growth, it is the target enzyme that fluorine class medicine acts on performance cytotoxicity in vivo simultaneously, fluorine class medicine is converted into 5-FU in vivo, the metabolite of 5-FU is combined with TS, hinder the normal function of TS, thereby it is synthetic to suppress DNA.Numerous clinical study results all show the mrna expression level of TYMS gene and fluorine class curative effect of medication closely related, it is better that the tumour patient of low TYMS mrna expression level is accepted the effect of fluorine class medicine chemotherapy, otherwise it is poor that the patient of TYMS high expression level accepts the effect of fluorine class medicine chemotherapy.
Above-mentioned result of study all shows, the expression amount that detects ERCC1, RRM1, TUBB3 and TYMS gene in the malignant tumor patient of first-line treatment has important clinical meaning, is the correct prerequisite to patient's medication treatment.
Past is mainly passed through Nothern blot method to the detection method of above-mentioned four gene expression amounts, the method need to be extracted RNA, use agarose gel electrophoresis, the RNA that molecular size range is different separate, subsequently its in-situ transesterification is moved on solid support, use again DNA or the rna probe of radioactivity (or on-radiation) mark, according to its homology, hybridize, finally carry out radioautograph (or chemical development), with target RNA position, indicate the size of its molecular weight, with the relative content of intensity prompting target RNA in institute's test sample product that develop.This method is loaded down with trivial details time-consuming, and RNA is had relatively high expectations, and the very easy degraded of RNA, causes unstable result, and sensitivity is not high, the very large guarantee detected result of sample rna amount needing.
Real-Time Fluorescent Quantitative PCR Technique is flourish at present, it refers in PCR reaction system and adds fluorophor, utilize the whole PCR process of fluorescent signal accumulation Real-Time Monitoring, finally by typical curve, unknown template is carried out the method for quantitative analysis, this technology has not only realized the detection by quantitative of PCR, and compare with conventional PCR, it has specificity and more by force, effectively solves PCR pollution problem, level of automation high.
Summary of the invention
In order to solve the problems of the technologies described above, the object of the present invention is to provide a kind of primer and probe for detection of ERCC1, RRM1, TUBB3 and TYMS gene expression amount, the invention also discloses the test kit and the using method thereof that include this primer, probe.
Present inventor retrieves the information of ERCC1, RRM1, TUBB3 and TYMS gene at GenBank, according to the gene coded sequence retrieving, conserved regions design primer and probe at gene, the two ends of its middle probe are mark fluorescent reporter group (R) and fluorescent quenching group (Q) respectively, by real-time fluorescence quantitative PCR, increase the expression of ERCC1, RRM1 in rapid detection sample to be tested, TUBB3 and TYMS gene, can detect accurately, rapidly and sensitively the expression amount of ERCC1, RRM1, TUBB3 and TYMS gene.
According to embodiments of the invention, a kind of primer and probe for detection of ERCC1, RRM1, TUBB3 and TYMS gene expression amount is provided, described primer sequence is as follows:
SEQ?ID?NO:15’-ATGGCCCTCACGGAGTTTTGT-3’
SEQ?ID?NO:25’-CATCTGGGCACTGAAAGGGA-3’
SEQ?ID?NO:35’-ACTAGCTGATGCATGTGA-3’
SEQ?ID?NO:45’-TCCATAACAAATTCTGGATTCG-3’
SEQ?ID?NO:55’-GCGGGAGGGATTTGGATT-3’
SEQ?ID?NO:65’-GGAAGGGTGGTTCCAGGC-3’
SEQ?ID?NO:75’-GAGGCGGTGGAGCACATTT-3’
SEQ?ID?NO:85’-CGTTTGTTGTGAGCAGAGG-3’
Described probe two ends are mark fluorescent reporter group and fluorescent quenching group respectively, and the sequence of described probe is as follows, and wherein, R represents fluorescence report group, and Q represents fluorescent quenching group:
SEQ?ID?NO:115’-R-ACGCCCTAGCTCACTCCCCT-Q-3’
SEQ?ID?NO:125’-R-GCCGCCAAGAAAGTCATGTTTGA-Q-3’
SEQ?ID?NO:135’-R-AAAGTTGTCTGGCTCCAA-Q-3’
SEQ?ID?NO:145’-R-CTGGCACCTGTCGGTGTT-Q-3’。
According to embodiments of the invention, a kind of test kit for detection of ERCC1, RRM1, TUBB3 and TYMS gene expression amount is also provided, it mainly comprises:
1) primer and the probe of ERCC1, RRM1, TUBB3 and TYMS gene test, wherein, primer sequence is as shown in SEQ ID NO:1～SEQ ID NO:8, and probe sequence is as shown in SEQ ID NO:11～SEQ ID NO:14;
2) primer of reference gene and probe; Preferably, reference gene is ATCB gene, and the primer sequence of this gene is as shown in SEQ ID NO:9 and SEQ ID NO:10, and probe sequence is as shown in SEQ ID NO:15, and described probe two ends are mark fluorescent reporter group and fluorescent quenching group respectively.
3) standard substance of ATCB, ERCC1, RRM1, TUBB3 and TYMS gene, the recombinant plasmid vector of described standard substance for containing ATCB, ERCC1, RRM1, TUBB3 and TYMS gene coded sequence respectively;
Wherein, the GenBank of ATCB gene coded sequence is numbered NM_001101.3, the GenBank of ERCC1 gene coded sequence is numbered NM_202001.2, the GenBank of RRM1 gene coded sequence is numbered NM_001033.3, the GenBank of TUBB3 gene coded sequence is numbered NM_001033.3, and the GenBank of TYMS gene coded sequence is numbered NM_021288.4;
4) real-time fluorescence quantitative PCR MIX reaction solution, deionized water;
5) separate and concentrate bottle or pipe and the packing box of packing mentioned reagent.
Wherein, the concentration of above-mentioned primer and probe is 5～10 μ mol/L.
Wherein, the empty carrier that contains ATCB, ERCC1, RRM1, TUBB3 and TYMS gene order plasmid vector described in step 3) is pGEM-T carrier.
Wherein, the real-time fluorescence quantitative PCR MIX reaction solution that described real-time fluorescence quantitative PCR MIX reaction solution is this area routine; Preferably, described real-time fluorescence quantitative PCR MIX reaction solution is the PrimeScript of Takara company
tMrT Master Mix.
The using method of primer of the present invention, probe and test kit, comprises the following steps:
(1) according to ordinary method, from testing sample, extract the total RNA of sample, and reverse transcription becomes cDNA;
(2) respectively by the ATCB of known copy number, ERCC1, RRM1, TUBB3 and TYMS gene standard substance gradient dilution to 10
6, 10
5, 10
4, 10
3, 10
2copy number/μ l;
(3) respectively with the cDNA of step (1) gained with take the gene standard substance of step (2) gradient dilution as template, add primer, probe, real-time fluorescence quantitative PCR MIX reaction solution, deionized water to set up reaction system;
Wherein, the primer sequence using for the detection of ERCC1 gene is as shown in SEQ ID NO:1 and SEQ ID NO:2, and probe sequence is SEQ ID NO:11 as shown;
The primer sequence using for the detection of RRM1 gene is as shown in SEQ ID NO:3 and SEQ ID NO:4, and probe sequence is SEQ ID NO:12 as shown;
The primer sequence using for the detection of TUBB3 gene is as shown in SEQ ID NO:5 and SEQ ID NO:6, and probe sequence is SEQ ID NO:13 as shown;
The primer sequence using for the detection of TYMS gene is as shown in SEQ ID NO:7 and SEQ ID NO:8, and probe sequence is SEQ ID NO:14 as shown;
The primer sequence using for reference gene ATCB is as shown in SEQ ID NO:9 and SEQ ID NO:10, and probe sequence is SEQ ID NO:15 as shown;
(4) use real-time fluorescence quantitative PCR instrument, according to routine operation method, carry out real-time fluorescence quantitative PCR amplification;
(5) analyzing and testing result: because the logarithm of the cycle threshold of template and the initial copy number of this template exists linear relationship, the initial copy number of each standard substance is taken the logarithm as X-coordinate, by its corresponding cycle threshold, it is ordinate zou, make corresponding typical curve, the initial copy number of the target gene according to the cycle threshold of cDNA to be measured, it being contained is again analyzed, and from typical curve, reads the initial copy number of its target gene containing.
By above technical scheme, beneficial effect of the present invention is as follows:
(1) can detect the expression amount of these four genes of ERCC1, RRM1, TUBB3 and TYMS, detect widely, truly reflect and the expression of these genes in patient body contribute to doctor that patient's pesticide application strategy is comprehensively analyzed, judged.
(2) testing process simple and fast, cheap, and detection sensitivity is high, can judge directly, rapidly and detected result obtain clinical required information in the very first time.
Embodiment
Below in conjunction with embodiment, the specific embodiment of the present invention is further described, advantage and disadvantage of the present invention will be more clear along with description.But these embodiment are only exemplary, scope of the present invention are not formed to any restriction.
The experimental technique using in following embodiment if no special instructions, is ordinary method.
In following embodiment, material used, reagent etc., if no special instructions, all can obtain from commercial channels.
Embodiment is for detection of test kit and the use thereof of ERCC1, RRM1, TUBB3 and TYMS gene expression amount
1. should in the test kit for detection of ERCC1, RRM1, TUBB3 and TYMS gene expression amount, be equipped with:
(1) for detection of primer and the probe of ERCC1, RRM1, TUBB3 and TYMS gene, primer sequence is as shown in SEQ ID NO:1～SEQ ID NO:8, the sequence of probe is as SEQ ID NO:11～SEQ ID NO:14, and described probe two ends are mark fluorescent reporter group and fluorescent quenching group respectively; The concentration of above-mentioned primer and probe is 5 μ mol/L.
(2) primer of reference gene ATCB and probe, primer sequence is as shown in SEQ ID NO:9 and SEQ ID NO:10, and probe sequence is as SEQ ID NO:15, and described probe two ends are mark fluorescent reporter group and fluorescent quenching group respectively; The concentration of above-mentioned primer and probe is 5 μ mol/L.
(3) standard substance of ATCB, ERCC1, RRM1, TUBB3 and TYMS gene, the recombinant plasmid vector of described standard substance for containing ATCB, ERCC1, RRM1, TUBB3 and TYMS gene coded sequence respectively; The empty carrier of described carrier is pGEM-T carrier.
Wherein, the GenBank of ATCB gene coded sequence is numbered NM_001101.3, the GenBank of ERCC1 gene coded sequence is numbered NM_202001.2, the GenBank of RRM1 gene coded sequence is numbered NM_001033.3, the GenBank of TUBB3 gene coded sequence is numbered NM_001033.3, and the GenBank of TYMS gene coded sequence is numbered NM_021288.4;
(4) real-time fluorescence quantitative PCR MIX reaction solution, deionized water; Described real-time fluorescence quantitative PCR Mix reaction solution is the PrimeScript of Takara company
tMrT Master Mix.
2. the using method of test kit is as follows:
(1) from paraffin section tissue sample, extract total RNA, and RNA reverse transcription is become to cDNA
The thin slice that paraffin-embedded tissue is cut into 5 – 10 μ m is placed in 1.5ml centrifuge tube, the dewaxing of use dimethylbenzene, add again dehydrated alcohol to process, room temperature is dried until residual ethanol has volatilized completely, with vacuum-drying instrument 45 degree, within 15 minutes, be dried, dry tissue is through protease K digesting and add Trizol again, and chloroform extracting obtains RNA, collect pure RNA and can directly be used for reverse transcription, also can be stored in-20 ℃ or-80 ℃;
Using total RNA of extracting as template, use random hexamers to become cDNA with reversed transcriptive enzyme reverse transcription.All operations all carries out on ice; In 0.2ml Eppendorf pipe, add following reaction system: total RNA5 μ g, random hexamers 1 μ l, RNase free water is to cumulative volume 12 μ l, and after centrifugal mixing, 70 ℃ of reactions 5 minutes, are placed on ice ice bath 5 minutes immediately; Add successively following composition, 5 * Reaction Buffer4 μ l, 10mM dNTP Mix2 μ l, RNA enzyme inhibitors (20u/ μ l) 1 μ l, slightly centrifugal each composition that mixes, 37 ℃ are reacted 5 minutes; Add reversed transcriptive enzyme (200u/ μ l) 1 μ l, 42 ℃ are reacted 60 minutes; 72 ℃ of 10 minutes deactivation reversed transcriptive enzymes.Be placed in rapidly ice bath 5 minutes, obtain cDNA ,-20 ℃ of preservations.
(2) respectively by the ATCB of known copy number, ERCC1, RRM1, TUBB3 and TYMS gene standard substance gradient dilution to 10
6, 10
5, 10
4, 10
3, 10
2copy number/μ l;
(3) respectively with the cDNA of step (1) gained with take the gene standard substance of step (2) gradient dilution as template, add primer, probe, real-time fluorescence quantitative PCR MIX reaction solution, deionized water to set up reaction system;
Wherein, the primer sequence using for the detection of ERCC1 gene is as shown in SEQ ID NO:1 and SEQ ID NO:2, and probe sequence is SEQ ID NO:11 as shown;
The primer sequence using for the detection of RRM1 gene is as shown in SEQ ID NO:3 and SEQ ID NO:4, and probe sequence is SEQ ID NO:12 as shown;
The primer sequence using for the detection of TUBB3 gene is as shown in SEQ ID NO:5 and SEQ ID NO:6, and probe sequence is SEQ ID NO:13 as shown;
The primer sequence using for the detection of TYMS gene is as shown in SEQ ID NO:7 and SEQ ID NO:8, and probe sequence is SEQ ID NO:14 as shown;
The primer sequence using for reference gene ATCB is as shown in SEQ ID NO:9 and SEQ ID NO:10, and probe sequence is SEQ ID NO:15 as shown;
Reaction system is as follows:
| Component | Volume (total) 20ul |
| 2XMaster?Mix | 10ul |
| Upstream primer | 1ul |
| Downstream primer | 1ul |
| Probe | 0.5ul |
| PCR water | 6.5ul |
| Template cDNA | 1ul |
The 7500PCR gene-amplificative instrament that uses ABI company, response procedures is:
95 ℃, 5min, 1 circulation;
95 ℃, 15s → 60 ℃, 60s, 40 circulations.
(4) analyzing and testing result: because the logarithm of the cycle threshold of template and the initial copy number of this template exists linear relationship, the initial copy number of each standard substance is taken the logarithm as X-coordinate, by its corresponding cycle threshold, it is ordinate zou, make corresponding typical curve, the initial copy number of the target gene according to the cycle threshold of cDNA to be measured, it being contained is again analyzed, and from typical curve, reads the initial copy number of its target gene containing.
It will be understood by those skilled in the art that lower without departing from the spirit and scope of the present invention and can the details of technical solution of the present invention and form be modified or be replaced, but these modifications and replacement all fall within the scope of protection of the present invention.
Claims (9)
1. primer and a probe, is characterized in that, the sequence of described primer is as follows:
SEQ?ID?NO:15’-ATGGCCCTCACGGAGTTTTGT-3’
SEQ?ID?NO:25’-CATCTGGGCACTGAAAGGGA-3’
SEQ?ID?NO:35’-ACTAGCTGATGCATGTGA-3’
SEQ?ID?NO:45’-TCCATAACAAATTCTGGATTCG-3’
SEQ?ID?NO:55’-GCGGGAGGGATTTGGATT-3’
SEQ?ID?NO:65’-GGAAGGGTGGTTCCAGGC-3’
SEQ?ID?NO:75’-GAGGCGGTGGAGCACATTT-3’
SEQ?ID?NO:85’-CGTTTGTTGTGAGCAGAGG-3’
Described probe two ends are mark fluorescent reporter group and fluorescent quenching group respectively, and the sequence of described probe is as follows, and wherein, R represents fluorescence report group, and Q represents fluorescent quenching group:
SEQ?ID?NO:115’-R-ACGCCCTAGCTCACTCCCCT-Q-3’
SEQ?ID?NO:125’-R-GCCGCCAAGAAAGTCATGTTTGA-Q-3’
SEQ?ID?NO:135’-R-AAAGTTGTCTGGCTCCAA-Q-3’
SEQ?ID?NO:145’-R-CTGGCACCTGTCGGTGTT-Q-3’。
2. a test kit that includes primer claimed in claim 1 and probe, is characterized in that, this test kit comprises:
1) primer and the probe of ERCC1, RRM1, TUBB3 and TYMS gene test, wherein, primer sequence is as shown in SEQ ID NO:1～SEQ ID NO:8, and probe sequence is as shown in SEQ ID NO:11～SEQ ID NO:14;
2) primer of reference gene and probe;
3) standard substance of reference gene, ERCC1, RRM1, TUBB3 and TYMS gene, the recombinant plasmid vector of described standard substance for containing reference gene, ERCC1, RRM1, TUBB3 and TYMS gene coded sequence respectively;
4) real-time fluorescence quantitative PCR MIX reaction solution, deionized water;
5) separate and concentrate bottle or pipe and the packing box of packing mentioned reagent.
3. according to test kit claimed in claim 2, it is characterized in that: described reference gene is ATCB gene, the primer sequence of described ATCB gene is as shown in SEQ ID NO:9 and SEQ ID NO:10, probe sequence is as shown in SEQ ID NO:15, and described probe two ends are mark fluorescent reporter group and fluorescent quenching group respectively.
4. according to test kit claimed in claim 2, it is characterized in that: the concentration of primer and probe is 5～10 μ mol/L.
5. according to test kit claimed in claim 2, it is characterized in that: the empty carrier that contains reference gene, ERCC1, RRM1, TUBB3 and TYMS gene order plasmid vector described in step 3) is pGEM-T carrier.
6. according to test kit claimed in claim 2, it is characterized in that: described real-time fluorescence quantitative PCR MIX reaction solution is the PrimeScript of Takara company
tMrT Master Mix.
7. the using method of primer and probe described in claim 1, comprises the following steps:
(1) according to ordinary method, from testing sample, extract the total RNA of sample, and reverse transcription becomes cDNA;
(2) respectively by the ATCB of known copy number, ERCC1, RRM1, TUBB3 and TYMS gene standard substance gradient dilution to 10
6, 10
5, 10
4, 10
3, 10
2copy number/μ l;
(3) respectively with the cDNA of step (1) gained with take the gene standard substance of step (2) gradient dilution as template, add primer, probe, real-time fluorescence quantitative PCR MIX reaction solution and deionized water to set up reaction system;
Wherein, the primer sequence using for the detection of ERCC1 gene is as shown in SEQ ID NO:1 and SEQ ID NO:2, and probe sequence is as shown in SEQ ID NO:11;
The primer sequence using for the detection of RRM1 gene is as shown in SEQ ID NO:3 and SEQ ID NO:4, and probe sequence is as shown in SEQ ID NO:12;
The primer sequence using for the detection of TUBB3 gene is as shown in SEQ ID NO:5 and SEQ ID NO:6, and probe sequence is as shown in SEQ ID NO:13;
The primer sequence using for the detection of TYMS gene is as shown in SEQ ID NO:7 and SEQ ID NO:8, and probe sequence is as shown in SEQ ID NO:14;
The primer sequence using for ATCB is as shown in SEQ ID NO:9 and SEQ ID NO:10, and probe sequence is as shown in SEQ ID NO:15;
(5) use real-time fluorescence quantitative PCR instrument, according to routine operation method, carry out real-time fluorescence quantitative PCR amplification;
(6) analyzing and testing result.
8. the application in detecting ERCC1, RRM1, TUBB3, TYMS gene expression amount according to primer claimed in claim 1 and probe.
9. the application in detecting ERCC1, RRM1, TUBB3, TYMS gene expression amount according to arbitrary described test kit in claim 2～6.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310537253.8A CN103602672B (en) | 2013-11-04 | 2013-11-04 | The primer of gene expression detection amount, probe and test kit and using method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310537253.8A CN103602672B (en) | 2013-11-04 | 2013-11-04 | The primer of gene expression detection amount, probe and test kit and using method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103602672A true CN103602672A (en) | 2014-02-26 |
| CN103602672B CN103602672B (en) | 2015-11-18 |
Family
ID=50120939
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310537253.8A Active CN103602672B (en) | 2013-11-04 | 2013-11-04 | The primer of gene expression detection amount, probe and test kit and using method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103602672B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111455055A (en) * | 2020-04-28 | 2020-07-28 | 重庆浦洛通基因医学研究院有限公司 | Human TYMS gene expression level detection standard reference substance |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012051800A1 (en) * | 2010-10-18 | 2012-04-26 | 北京雅康博生物科技有限公司 | Plasmid standard for use in quantitative assays using fluorescent quantitative pcr |
| WO2012092336A2 (en) * | 2010-12-28 | 2012-07-05 | Caris Mpi, Inc. | Molecular profiling for cancer |
-
2013
- 2013-11-04 CN CN201310537253.8A patent/CN103602672B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012051800A1 (en) * | 2010-10-18 | 2012-04-26 | 北京雅康博生物科技有限公司 | Plasmid standard for use in quantitative assays using fluorescent quantitative pcr |
| CN102453748A (en) * | 2010-10-18 | 2012-05-16 | 北京雅康博生物科技有限公司 | Plasmid standard substance for quantitative detection by fluorescence quantitative PCR |
| WO2012092336A2 (en) * | 2010-12-28 | 2012-07-05 | Caris Mpi, Inc. | Molecular profiling for cancer |
Non-Patent Citations (2)
| Title |
|---|
| 伍青等: "多靶标检测平台分析肺腺癌患者不同分化程度中ERCC1、TYMS、RRM1、TUBB3 基因表达及意义", 《现代肿瘤医学》 * |
| 孟庆雨等: "ERCC1、TYMS、RRM1、TUBB3 表达与非小细胞肺癌化疗疗效的相关性和意义", 《中国肿瘤》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111455055A (en) * | 2020-04-28 | 2020-07-28 | 重庆浦洛通基因医学研究院有限公司 | Human TYMS gene expression level detection standard reference substance |
| CN111455055B (en) * | 2020-04-28 | 2021-11-16 | 重庆浦洛通基因医学研究院有限公司 | Human TYMS gene expression level detection standard reference substance |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103602672B (en) | 2015-11-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Giannopoulou et al. | Liquid biopsy in ovarian cancer: the potential of circulating miRNAs and exosomes | |
| Szafranska et al. | MicroRNA expression alterations are linked to tumorigenesis and non-neoplastic processes in pancreatic ductal adenocarcinoma | |
| Qu et al. | Circulating miRNAs: promising biomarkers of human cancer | |
| Hong et al. | MicroRNA expression profiling of diagnostic needle aspirates from surgical pancreatic cancer specimens | |
| Bournet et al. | Role of endoscopic ultrasound in the molecular diagnosis of pancreatic cancer | |
| Usó et al. | miRNA detection methods and clinical implications in lung cancer | |
| CN104328164A (en) | Kit for detecting human EGFR gene mutation by using fluorescence probe hybridization method | |
| CN107312852A (en) | Myocardial infarction diagnosis mark compositions | |
| Segersten et al. | The role of microRNA profiling in prognosticating progression in Ta and T1 urinary bladder cancer | |
| CN101613748A (en) | A kind of method that detects serum marker of pancreatic cancer | |
| CN105420393A (en) | Primers, probe, and kit for detecting BRCA1 gene expression | |
| Zhou et al. | Clinical impact of serum miR-661 in diagnosis and prognosis of non-small cell lung cancer. | |
| Zhang et al. | Correlations of the MiR-330 expression with the pathogenesis and prognosis of breast cancer. | |
| CN107299129B (en) | Application of circulating nucleic acid as breast cancer biomarker | |
| US20150247202A1 (en) | Microrna based method for diagnosis of colorectal tumors and of metastasis | |
| CN103602672B (en) | The primer of gene expression detection amount, probe and test kit and using method thereof | |
| US20160208346A1 (en) | Method and composition for detection of oncogenic hpv | |
| CN112813167B (en) | Marker LINC01977 and application thereof | |
| CN105154560B (en) | PCR kit for fluorescence quantitative and detection method and purposes for oophoroma | |
| CN113832232A (en) | Biomarker for predicting responsiveness of lung adenocarcinoma to platinum-containing dual-drug chemotherapy, and related product | |
| Zhang et al. | Increased expression of long non-coding RNA GLIDR in prostate cancer | |
| CN103589786B (en) | Method, test kit and the primer of detection RRM1 mRNA relative expression quantity and probe | |
| CN103725787B (en) | Fluorescent PCR (Polymerase Chain Reaction) detection kit for fusion genes of leukemia | |
| CN206359543U (en) | A kind of kit for being used to detect the first-line drug expression of drug resistance genes amount for the treatment of malignant tumour | |
| CN102424843B (en) | Application and detection kit of human miR-183/96/182 cluster |
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 | ||
| CP03 | Change of name, title or address | ||
| CP03 | Change of name, title or address |
Address after: 100176 building 4, building 3, No. 1, Desheng North Street, Beijing Economic and Technological Development Zone, Daxing District, Beijing Patentee after: Beijing Hester medical laboratory Co. Ltd. Address before: 100176 A-3, A-4, 1 North Sheng street, Yizhuang economic and Technological Development Zone, Beijing, Daxing District Patentee before: BEIJING HAISITE CLINICAL INSPECTION CO., LTD. |