CN105400873A - Primer, probe and kit for fluorescently and quantitatively detecting TOP2A gene expression through PCR method - Google Patents
Primer, probe and kit for fluorescently and quantitatively detecting TOP2A gene expression through PCR method Download PDFInfo
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Abstract
The invention discloses a primer, probe and kit for fluorescently and quantitatively detecting TOP2A gene expression through a PCR method. The nucleotide sequences of a primer pair A for fluorescently and quantitatively detecting mRNA of a TOP2A gene are shown in SEQ ID NO:1 and SEQ ID NO:2. The nucleotide sequence of the Taqman probe A is shown in SEQ ID NO:3. The kit comprises a primer for detection and a probe for detection, and preferentially further comprises an mRNA detection primer and probe of a reference gene B2M, a first chain cDNA synthesis system, fluorescent and quantitative PCR detection reaction liquid, a positive standard product, a reference standard product, a negative contrast product and the like. The primer, probe and kit have the advantages of being high in specificity, sensitivity, repeatability and the like and have the advantages of being accurate in quantification, capable of premixing enzymes, high in speed, capable of achieving the fully-closed reaction and the like, and the problems that an existing detecting method is long in time, poor in accuracy, prone to pollution and the like can be effectively solved.
Description
Technical field
The present invention relates to molecular biology, biotechnology and clinical molecular diagnosis technical field, particularly relate to the primer of fluorescence quantitative PCR method detection TOP2A gene mRNA expression, probe and test kit.
Background technology
Mammary cancer is one of common malignant tumour of women, occupies first of tumour at its sickness rate of western countries and case fatality rate.In recent years, in the trend that China's sickness rate also linearly rises, current infiltrative breast carcinoma is regarded as systemic disease, judgement and the biological markers relevant to prognosis of its prognosis receive much concern, due to the subclinical metastasis of many patients, and the appearance of chemotherapy resistance, cause patient's Sulfurless fixative to shorten, detect relevant breast carcinoma resistance genes involved, particularly important to instructing Normalized Treatment.
Find in the research of mammary cancer, genetic expression and the susceptibility of tumour cell to Topoisomerase inhibitors of TOPO II α are obviously correlated with, the patient of TOP2A gene unconventionality obviously can benefit from the adjuvant chemotherapy scheme containing Topoisomerase inhibitors, the patient of TOP2A high expression level uses CEF scheme to carry out treatment can reduce the risk of recurrence of 61% and the mortality risk of 51%, and the patient of the low expression of TOP2A uses CEF scheme can only reduce the risk of recurrence of 6% and the mortality risk of 10%.
TOP2A gene (Topoisomerase II alpha, TOP II α) be positioned 17q11.2-22 region, coding DNA topoisomeraseⅡ α, it is one of nuclear matrix elements, play a role in core, the dynamic change of nucleic acid space structure can be regulated, participate in the copying of DNA, transcribe, recombinate and repair process.
Etoposide is cell cycle specific antitumor drug, acts on TOP-001, forms the reversibility mixture that medicine-enzyme-DNA is stable, hinders DNA to repair.Research finds, the susceptibility of Etoposide depends on the expression level of its cell TOPO II α albumen.The patient of TOPO II α high expression level can benefit from Etoposide treatment.Therefore TOP2A gene mRNA levels is used to pass judgment on patient to anthracycline-containing therapeutic scheme, more accurately.
Mainly contain immunohistochemical method for the method detecting TOP2AmRNA level at present, the method is long for detection time, and interpretation affects greatly by subjective factor, accurately can not reflect the expression level of TOP2A; And there is the problems such as sensitivity is low, easy pollution in cDNA microarray mrna expression spectrum analysis (gene chip) method.
Summary of the invention
In order to solve the aforementioned problems in the prior, the invention provides primer pair and Taqman probe that a kind of fluorescent quantitation detects TOP2A gene mRNA, high specificity, highly sensitive, reproducible.
Fluorescent quantitation provided by the invention detects primer pair A and the Taqman probe A of TOP2A gene mRNA, and the nucleotide sequence of its primer pair A is as shown in SEQIDNO:1 ~ 2, and the nucleotide sequence of Taqman probe A is as shown in SEQIDNO:3.
The present invention adopts fluorescent PCR in conjunction with Taqman probe method, the gene expression dose of TOP2A is detected based on fluorescent quantitative PCR technique, devise cDNA amplimer and the specificity T aqman probe of goal gene, natch, 5 ' end of Taqman probe is marked with fluorescent reporter group, and 3 ' end is marked with fluorescent quenching group.The preferred reporter group of the present invention is FAM, and quenching group is MGB.The nucleotide sequence of primer and probe is specifically:
Primer pair A:F:5 '-AAGTTACCTGAAGCTC-3 ' (SEQIDNO:1);
R：5’-GGAAGGTGTTTTTAG-3’(SEQIDNO:2)；
Taqman probe A:5 ' FAM-TCCCCTCTGAATT-MGB3 ' (SEQIDNO:3).
Present invention also offers the test kit that a kind of fluorescence quantitative PCR method detects TOP2A genetic expression, it comprises primer pair A and Taqman probe A that above-described fluorescent quantitation detects TOP2A gene.
Preferably, in mentioned reagent box, also comprise the primer pair B for detecting reference gene B2MmRNA and Taqman probe B, the nucleotide sequence of primer pair B is as shown in SEQIDNO:4 ~ 5, and the nucleotide sequence of Taqman probe B is as shown in SEQIDNO:6.The reporter group of preferred Taqman probe B is FAM, and quenching group is MGB.The object of selection reference gene B2M is the accuracy in order to increase detection.Compared to other reference gene, experiment shows, B2M is that differential expression in the tumour of internal reference and normal group and between group is minimum compared with GAPDH and GUSB etc., optimal stability.
Primer pair B:F:5 '-ATGCTTATACACTTACA-3 ' (SEQIDNO:4),
R:5’-ACATGGACATGATCTTC-3’(SEQIDNO:5)；
Taqman probe B:5 ' FAM-GTAGGGTTATAATA-MGB3 ' (SEQIDNO:6).
Preferably, in mentioned reagent box, also comprise B2M standard substance, TOP2A standard substance.TOP2A standard substance are the solution containing gradient concentration TOP2A geneome plasmid DNA, and B2M standard substance are the solution containing gradient concentration B2M geneome plasmid DNA, and standard substance are the positive control as this test kit and the making for typical curve.
Preferably, in mentioned reagent box, also comprise the first chain cDNA synthetic system, specifically comprise that ThermoScript II 1 ~ 5U, reverse transcription 10 × RTBuffer are appropriate, dNTPs1 ~ 2mM, TOP2A gene reverse transcription primer to 1 ~ 10pM, complement to 19.5 μ Ls with DEPC water to 1 ~ 10pM, reference gene B2M reverse transcription primer; Sample rna 0.5 μ g is added during use.Wherein the reverse transcription primer of two genes can designed, designed as required, or directly buys commodity, as long as can obtain the cDNA of its correspondence, without other requirements.10 × RTBuffer is reverse transcription PCR damping fluid, directly can buy commercial goods.
Preferably, have five pipe reagent in mentioned reagent box, be respectively:
First chain cDNA synthetic system;
Fluorescence quantitative PCR detection reaction solution: containing Taq DNA polymerase 1 ~ 2U, 2.0 ~ 5.0mmolMgCl
2with the Premix of 0.2 ~ 0.8mmoldNTPs; Primer pair A1 ~ 10pM; Taqman probe A1 ~ 10pM; Primer pair B1 ~ 10pM; Taqman probe B1 ~ 10pM; DEPC water is appropriate;
TOP2A standard substance;
B2M standard substance;
Negative controls: DEPC water.
A and B of the present invention, only for distinguishing different primer pairs and probe, does not have other special significances.
Present invention also offers the using method that above arbitrary described fluorescence quantitative PCR method detects the test kit of TOP2A genetic expression, it comprises the following steps:
(1) with tumor tissues and Carcinoma side normal tissue for measuring samples, extract the RNA in measuring samples, reverse transcription becomes sample cDNA;
(2) using sample cDNA as template, quantitative fluorescent PCR reaction is carried out with primer pair A, Taqman probe A, primer pair B and Taqman probe B; TOP2A standard substance and B2M standard substance also carry out the operation same with sample cDNA respectively, and according to TOP2A standard substance and B2M standard substance drawing standard curve;
(3) interpretation of result: the R of analytical standard curve
2more than 0.99, TOP2A gene and B2M gene amplification efficiency in 90-100% scope, then according to the Ct value that quantitative fluorescent PCR instrument provides, should calculate as follows:
△ Ct (tumor tissues)=Ct (neoplasmic tissue sample to be checked)-Ct (B2M gene);
△ Ct (Carcinoma side normal tissue)=Ct (Carcinoma side normal tissue sample to be checked)-Ct (B2M gene);
△ △ Ct=△ Ct (tumor tissues)-△ Ct (Carcinoma side normal tissue);
Relative expression leads=and 2
-△ △ Ct; Relative expression leads > 1 and is judged as mammary cancer TOP2A gene high expression.
Preferably, in above-mentioned using method, in step (2), quantitative fluorescent PCR response procedures is: 95 DEG C of denaturation 2min; 95 DEG C of 15s, 60 DEG C 30s40 circulation; Fluorescent signal is gathered time when fluorophor selects FAM at 60 DEG C of 30s.
Compared with prior art, the present invention has following beneficial effect:
Fluorescent quantitation provided by the invention detects primer pair and the Taqman probe of TOP2A gene mRNA, high specificity, highly sensitive, reproducible.Based on this fluorescence quantitative detection kit, also there is high specificity, the feature such as highly sensitive, reproducible, also have quantitatively accurately simultaneously, the advantage such as premix enzyme, speed are fast, totally-enclosed reaction:
1, operability: easy and simple to handle, a step application of sample, without the need to being mixed with enzyme by sample, only needs 2 hours from sample extraction to going out examining report.
2, sensitivity: this test kit can carry out limited amplification to the sample of 1ng, and detected result is credible.
3, specificity: the upstream and downstream primer of design is Auele Specific Primer, uses Taqman probe method to carry out quantitatively, having very high specificity.
4, accuracy: using healthy tissues as quantitative medium, synchronous PCR reaction process, measurement under equal experiment condition realizes Ct value and mutually compares, and can reduce the error that experimentation causes, and the mRNA level in-site realizing fluorescence quantitative PCR detection TOP2A gene detects.
5, range of application: of the present inventionly can detect Fresh Frozen tissue, paraffin embedding sample, blood, have higher sensitivity and specificity, the clinical scope of application is wide.
Accompanying drawing explanation
Fig. 1: TOP2A canonical plotting;
Fig. 2: B2M canonical plotting;
Fig. 3: TOP2A low expression amplification curve diagram.
Embodiment
Below in conjunction with preferred specific embodiment, the invention will be further described, and to make those skilled in the art the present invention may be better understood and can be implemented, but illustrated embodiment is not as a limitation of the invention.
Embodiment 1: the preparation of test kit of the present invention
Test kit of the present invention is composed as follows:
1. the preparation of the first chain cDNA synthetic system is in table 1, the reaction system of 19.5 μ L, the about 0.5 μ g of sample RNA consumption.
Table 1
2. the preparation of fluorescence quantitative PCR detection reaction solution is in table 2 (system of 20 μ L, the about 5 μ L of cDNA template consumption):
Table 2
| Reagent | Consumption |
| Premix (autogamy) | 1.5μL |
| Primer pair A, primer pair B | Often couple of each 1 ~ 10pM |
| Taqman probe A, Taqman probe B | Each 1 ~ the 10pM of every bar |
| Mend DEPC water extremely | 20μL |
3. TOP2A standard substance (positive criteria product): the solution containing TOP2A geneome plasmid DNA, wherein TOP2A geneome plasmid DNA concentration is respectively: 3 × 10
5copy/microlitre, 6 × 10
4copy/microlitre, 1.2 × 10
4copy/microlitre, 2.4 × 10
3copy/microlitre, 4.8 × 10
2copy/microlitre.
4. B2M standard substance (internal reference standard substance): the solution containing B2M geneome plasmid DNA, wherein B2M geneome plasmid DNA concentration is respectively: 3 × 10
5copy/microlitre, 6 × 10
4copy/microlitre, 1.2 × 10
4copy/microlitre, 2.4 × 10
3copy/microlitre, 4.8 × 10
2copy/microlitre.
5. negative controls: contrast with DEPC water belongs with yin.
Embodiment 2: the expression amount detecting TOP2AmRNA with the test kit of preparation in embodiment 1
The present embodiment extracts RNA from the normal sample slice of my company 40 routine clinical breast cancer paraffin-embedded tissue sample and correspondence, and carry out quantitatively to it, as the template that PCR detects.Concrete operation step extracts test kit with reference to QIAGENRNeasyFFPEKit paraffin-embedded tissue RNA extraction agent box specification sheets, extracts sample rna.The RNA extracted calculates its purity and concentration quantitative through ultraviolet spectrophotometer, with DEPC water, the RNA of extracting is diluted to same concentrations, concentration 10ng/ μ L.
Detection method is as follows:
(1) using the first chain cDNA synthetic system, is cDNA by RNA reverse transcription.First chain reverse transcription program is: 50 DEG C, 30min; 95 DEG C, 5min; 5 DEG C, 5min1 circulation.
(2) cDNA10 of the healthy tissues of 40 routine mammary cancer and correspondence is doubly diluted be placed in stand-by on ice.
(3) (concentration of TOP2A geneome plasmid DNA is respectively 3 × 10 inside test kit, to take out different concns positive criteria product
5, 6 × 10
4, 1.2 × 10
4, 2.4 × 10
3, 4.8 × 10
2copy/microlitre); (B2M geneome plasmid DNA is respectively 3 × 10 to different concns internal reference standard substance
5, 6 × 10
4, 1.2 × 10
4, 2.4 × 10
3, 4.8 × 10
2copy/microlitre) and negative controls, shake evenly centrifugal, be placed in stand-by on ice.
(4) the TOP2A standard substance of 5 different concns gradients, tumor sample cDNA, the other normal cDNA of cancer, negative controls are joined in goal gene TOP2APCR reaction solution respectively; Separately the B2M standard substance of 5 different concns gradients, tumor sample cDNA, the other normal cDNA of cancer, negative controls are joined in house-keeping gene B2MPCR reaction solution respectively; The equal 5 μ L/ holes of two reaction systems.
(5) PCR response procedures: 95 DEG C of denaturation 2min; 95 DEG C of 15s, 60 DEG C 30s40 circulation, fluorescent signal FAM gathers when 60 DEG C of 30s.
(6) interpretation of result: the R2 of analytical standard curve is more than 0.99, and goal gene and house-keeping gene amplification efficiency should in 94-100% scopes, then according to the Ct value that quantitative fluorescent PCR instrument provides:
△ Ct (tumor tissues)=Ct (neoplasmic tissue sample to be checked)-Ct (B2M gene);
△ Ct (Carcinoma side normal tissue)=Ct (Carcinoma side normal tissue sample to be checked)-Ct (B2M gene);
△ △ Ct=△ Ct (tumor tissues)-△ Ct (Carcinoma side normal tissue)
Relative expression leads=and 2
-△ △ Ct; Relative expression leads > 1 and is judged as mammary cancer TOP2A gene high expression.The relative expression quantity data results of the TOP2A of 40 routine breast cancer case is as follows:
The relative expression quantity of TOP2A is adopted to be 1 as threshold value, the high expression level of mammary cancer when being greater than 1, be normal expression when being less than or equal to 1, this test kit, as 85%, can be used for clinically to the judgement of mammary cancer whether high expression level by the result coincidence rate that this result judges for cancerous tissue and healthy tissues thereof.
Table 3: kit results judges reference value
| Relative expression leads | Judge |
| 2 -△△Ct＞1 | High expression level |
| 2 -△△Ct≤1 | Low expression |
The above embodiment is only that protection scope of the present invention is not limited thereto in order to absolutely prove the preferred embodiment that the present invention lifts.The equivalent alternative or conversion that those skilled in the art do on basis of the present invention, all within protection scope of the present invention.Protection scope of the present invention is as the criterion with claims.
SEQUENCELISTING
<110> Wuhan Hygiea Bioscience Co., Ltd.
<120> fluorescence quantitative PCR method detects the primer of TOP2A genetic expression, probe and test kit
<130>
<160>6
<170>PatentInversion3.3
<210>1
<211>16
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<213> artificial sequence
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ggaaggtgtttttag15
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tcccctctgaatt13
<210>4
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atgcttatacacttaca17
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gtagggttataata14
Claims (9)
1. fluorescent quantitation detects primer pair A and the Taqman probe A of TOP2A gene mRNA, and it is characterized in that, the nucleotide sequence of primer pair A is as shown in SEQIDNO:1 ~ 2, and the nucleotide sequence of Taqman probe A is as shown in SEQIDNO:3.
2. fluorescent quantitation according to claim 1 detects primer pair A and the Taqman probe A of TOP2A gene, it is characterized in that, the nucleotide sequence 5 ' of Taqman probe A holds the fluorescent reporter group of mark to be FAM, and the quenching group of 3 ' end mark is MGB.
3. fluorescence quantitative PCR method detects a test kit for TOP2A genetic expression, it is characterized in that, the fluorescent quantitation comprised described in claim 1 or 2 detects primer pair A and the Taqman probe A of TOP2A gene.
4. fluorescence quantitative PCR method according to claim 3 detects the test kit of TOP2A genetic expression, it is characterized in that, also comprise the primer pair B for detecting reference gene B2MmRNA and Taqman probe B, the nucleotide sequence of primer pair B is as shown in SEQIDNO:4 ~ 5, and the nucleotide sequence of Taqman probe B is as shown in SEQIDNO:6.
5. fluorescence quantitative PCR method according to claim 4 detects the test kit of TOP2A genetic expression, it is characterized in that, also comprise TOP2A standard substance, B2M standard substance and negative controls, TOP2A standard substance are the solution containing gradient concentration TOP2A geneome plasmid DNA, and B2M standard substance are the solution containing gradient concentration B2M geneome plasmid DNA.
6. fluorescence quantitative PCR method according to claim 5 detects the test kit of TOP2A genetic expression, it is characterized in that, also comprise the first chain cDNA synthetic system, specifically comprise that ThermoScript II 1 ~ 5U, reverse transcription 10 × RTBuffe are appropriate, dNTPs1 ~ 2mM, TOP2A gene reverse transcription primer to 1 ~ 10pM, complement to 19.5 μ Ls with DEPC water to 1 ~ 10pM, reference gene B2M reverse transcription primer; Sample rna 0.5 μ g is added during use.
7. fluorescence quantitative PCR method according to claim 6 detects the test kit of TOP2A genetic expression, it is characterized in that, has five pipe reagent, be respectively in test kit:
First chain cDNA synthetic system;
Fluorescence quantitative PCR detection reaction solution: containing Taq DNA polymerase 1 ~ 2U, 2.0 ~ 5.0mmolMgCl
2with the Premix of 0.2 ~ 0.8mmoldNTPs; Primer pair A1 ~ 10pM; Taqman probe A1 ~ 10pM; Primer pair B1 ~ 10pM; Taqman probe B1 ~ 10pM; DEPC water is appropriate;
TOP2A standard substance;
B2M standard substance;
Negative controls: DEPC water.
8. the arbitrary described fluorescence quantitative PCR method of claim 4 ~ 7 detects the using method of the test kit of TOP2A genetic expression, it is characterized in that, comprises the following steps:
(1) with tumor tissues and Carcinoma side normal tissue for measuring samples, extract the RNA in measuring samples, reverse transcription becomes sample cDNA;
(2) using sample cDNA as template, quantitative fluorescent PCR reaction is carried out with primer pair A, Taqman probe A, primer pair B and Taqman probe B; TOP2A standard substance and B2M standard substance also carry out the operation same with sample cDNA respectively, and according to TOP2A standard substance and B2M standard substance drawing standard curve;
(3) interpretation of result: the R of analytical standard curve
2more than 0.99, TOP2A gene and B2M gene amplification efficiency in 94-100% scope, then according to the Ct value that quantitative fluorescent PCR instrument provides, should calculate as follows:
△ Ct (tumor tissues)=Ct (neoplasmic tissue sample to be checked)-Ct (B2M gene);
△ Ct (Carcinoma side normal tissue)=Ct (Carcinoma side normal tissue sample to be checked)-Ct (B2M gene);
△ △ Ct=△ Ct (tumor tissues)-△ Ct (Carcinoma side normal tissue);
relative expression leads > 1 and is judged as mammary cancer TOP2A gene high expression.
9. fluorescence quantitative PCR method according to claim 8 detects the using method of the test kit of TOP2A genetic expression, it is characterized in that, in step (2), quantitative fluorescent PCR response procedures is: 95 DEG C of denaturation 2min; 95 DEG C of 15s, 60 DEG C 30s40 circulation; Fluorescent signal is gathered time when fluorophor selects FAM at 60 DEG C of 30s.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107475372A (en) * | 2017-07-23 | 2017-12-15 | 嘉兴允英医学检验有限公司 | A kind of kit for CTLA 4mRNA detection of expression |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101698884A (en) * | 2009-11-10 | 2010-04-28 | 广州益善生物技术有限公司 | Detection liquid-phase chip and detection method for expression level of mRANs of TOP2A genes |
| CN103882138A (en) * | 2014-04-04 | 2014-06-25 | 上海赛安生物医药科技有限公司 | Multiple-gene detecting kit related to antitumor drugs |
-
2015
- 2015-11-24 CN CN201510824467.2A patent/CN105400873A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101698884A (en) * | 2009-11-10 | 2010-04-28 | 广州益善生物技术有限公司 | Detection liquid-phase chip and detection method for expression level of mRANs of TOP2A genes |
| CN103882138A (en) * | 2014-04-04 | 2014-06-25 | 上海赛安生物医药科技有限公司 | Multiple-gene detecting kit related to antitumor drugs |
Non-Patent Citations (6)
| Title |
|---|
| HUIJUAN WANG ET AL.: "Tissue-specific selection of optimal reference genes for expression analysis of anti-cancer drug-related genes in tumor samples using quantitative real-time RT-PCR", 《EXPERIMENTAL AND MOLECULAR PATHOLOGY》 * |
| 刘春萍等: "乳腺癌Her-2与TOP2A基因表达对新辅助化疗效果的预测价值", 《中国普通外科杂志》 * |
| 杜娟: "《医学细胞与分子生物学理论与技术》", 31 July 2012, 吉林大学出版社 * |
| 胡新文等: "《热带植物基因工程原理与操作技术》", 31 December 2006, 中国林业出版社 * |
| 董恩妮等: "实时荧光定量PCR内参基因的选择", 《中国畜牧杂志》 * |
| 贾玉萍: "Real-time PCR技术检测乳腺癌患者HER2和TOP2A基因的表达及相关性研究", 《临床医药实践》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107475372A (en) * | 2017-07-23 | 2017-12-15 | 嘉兴允英医学检验有限公司 | A kind of kit for CTLA 4mRNA detection of expression |
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Application publication date: 20160316 |