CN103436594A - Application of BDNA for detecting PCA3 in urine - Google Patents
Application of BDNA for detecting PCA3 in urine Download PDFInfo
- Publication number
- CN103436594A CN103436594A CN2012103914886A CN201210391488A CN103436594A CN 103436594 A CN103436594 A CN 103436594A CN 2012103914886 A CN2012103914886 A CN 2012103914886A CN 201210391488 A CN201210391488 A CN 201210391488A CN 103436594 A CN103436594 A CN 103436594A
- Authority
- CN
- China
- Prior art keywords
- pca3
- urine
- prostate cancer
- probe
- bdna
- 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
Abstract
The invention provides an application of BDNA for detecting PCA3 in urine. Operation steps comprise (1) collecting PCA3 specimens; (2) studying the collection of the PCA3 specimens; (3) detecting with a BDNA technology. By utilizing the bDNA technology being good at capture of small fragment genes, efficiently specific combination of uniquely designed target probes and PCA3 and an improved signal amplifying system, both sensitivity and specificity are guaranteed. The technology prevents the problems of false positive and false negative of conventional PCR, is simple in operations, and can be operated by a person with basic skills of molecular biology experiments. Meanwhile, the technology has high detected result sensitivity, high positive detectable rate and good stability and repeatability, is convenient for operations, is time-saving and can be popularized easily.
Description
Technical field
The present invention relates to the prostate cancer detection field, relate in particular to a kind of BDNA and detect the application of PCA3 in urine.
Background technology
At present, prostatic cancer early diagnosis method and PCA3 advantage prostate cancer (PCa) are the common cancers of the western countries male sex, are positioned at cause of death second.Along with the raising of China's aging population, dietary structure west and diagnostic techniques, sickness rate and the diagnosis of PCa increase year by year.
The most frequently used marker of diagnosing prostate cancer is PSA, but PSA is the prostata tissue specific antigens, be not the PCa specific antigens, therefore benign prostatic hyperplasia, prostatitis, acute urinary retention, prostatic various operation techniques and inspection all can cause that blood-serum P SA is high, disease specific is poor.In the elderly men of every 4 the blood-serum P SA>4ng/ml of the U.S. only 1 people be punctured biopsy and turn out to be prostate cancer.15% the PCa patients serum PSA of also having an appointment is less than 4ng/mL.Clinically in the urgent need to a kind of tumour-specific markers thing, carry out diagnosing prostate cancer.And the PCA3 gene is the PCa specific gene, therefore use the PCA3 gene will better susceptibility and specificity be arranged than PSA as the tumor marker of diagnosis PCa.
PCA3 is positioned Chromosome 9 (9q21-22), and the about 25kb of total length comprises 3 introns and 4 exons.Research shows that PCA3 is a kind of non-coding RNA, and its protein product is considerably less, therefore the mRNA detection of PCA3 is had more to specific aim.
PCA3 through very long research, predicts that to PCA3 susceptibility and the specificity of prostate cancer are sure abroad at present after within 1999, being found first, and it is better than PSA.The specificity overexpression of PCA3 in prostate cancer tissue makes it become the prostate cancer marker that one prospective is wide.But to mechanism, its biological function and the PCA3 of PCA3 high expression level prostate cancer form and the crucial biochemical route of progression of disease in have the aspect such as what meaning still exist many unknowns, need the field of exploring.Aspect diagnosing tumor, because the PCA3 transcription product is only the fragment of tens kb, and therefore mRNA easily degraded in general environment and transportation are preserved adopts any technology platform to detect PCA3 and guarantees that diagnosis is crucial accurately and reliably.The external PCA3 that detects mainly contains three kinds of modes, round pcr, TMA technology and bDNA technology at present.First two technology maximum uncertainty is intrinsic false positive and the Problem of False Negative that amplification of nucleic acid sequences brings.Because the amplification of this technology is to use primer amplification, it is complete needing the zone of amplification, as the RNA in sample ruptures and degrades or make a variation, just can't be combined with primer, causes false negative.BDNA technology (branched DNA, bDNA) be current unique state-of-the-art technology detected for virus of AIDS (HIV) and hepatitis C virus (HCV) by the U.S. FDA approval, the probe used in the bDNA technology comprises target-probe (target probe), front amplification body (preamplifier), amplification body (amplifier), label probe (label probe).In above-mentioned probe, only have target-probe to carry out different designs for different goal gene, rear three kinds of probes are all general to the bDNA analysis of any gene.Target-probe has comprised the multiple specific probe for PCA3 gene different fragments, for the captured target gene fragment.Amplifying probe is the key of technology.
Summary of the invention
Purpose of the present invention is for solving above technical problem, provide a kind of highly sensitive, positive rate is high, stability and favorable repeatability, easy and simple to handle, save time, be easy to popularize detect the application of PCA3 in urine.
In order to solve the problems of the technologies described above, the application that BDNA provided by the invention detects PCA3 in urine is achieved in that
A kind of BDNA detects the application of PCA3 in urine, and the operation steps adopted is:
(1) the PCA3 sample is collected
Urine PCA3 detects the urine specimen after using sample for the per rectum massage of prostate:
1) check that front drinking-water 500ml contributes to collect urine specimen;
2) prostate gland refers to collect immediately urine after inspection;
3) at least wanting the 20-30ml urine, be collected in the cup that posts label, must be the urine specimen of initial excretion, in the urine cup of preservation urine, should not contain any sanitas;
4), if the sample prolonged preservation should be put-80 ℃, detect in a short time and place-20 ℃ of preservations.
(2) research PCA3 sample is collected
1) research and utilization PCA3 as target the feasibility in prostate cancer diagnosis individuation assessment;
2) research bDNA technology platform detects after the per rectum massage of prostate sensitivity, specificity, the detection speed of PCA3 scoring diagnosing prostate cancer in urine, and whether is convenient to flux and detects;
3) the patients with prostate cancer PCA3 scoring of different clinical stagess of research and pathological grading changes whether significant difference and clinical meaning are arranged;
4) research PCA3 scoring, as the feasibility of monitoring index after prostate cancer progress and prognosis judging quota and prostate cancer therapy;
5) the ROC curve of Study of China people PCA3 and standards of grading and diagnostic threshold;
6) research PCA3 isomer is in Chinese patients with prostate cancer crowd's expression and whether variant expression in the tumor specimen of different pathological classification;
7) under research PCA3 different isomerization body surface reaches, androgen receptor isomer expression, whether analyze both has dependency.
(3) utilize the BDNA technology for detection:
1) set up the optimum detection flow process with PCA3 early diagnosis prostate gland cancer in bDNA technology for detection urine;
2) set up ROC curve and the PCA3 diagnostic threshold of Chinese PCA3 scoring, and PCA3 is to the evaluation system in the prostate cancer Individual Diagnosis; And the mark using PCA3 as the Noninvasive diagnosis prostate cancer is at clinical application;
3) understand Chinese patients with prostate cancer PCA3 isomer expression specificity;
4) understand Chinese patients with prostate cancer PCA3 isomer and express the dependency of expressing with the androgen receptor isomer.
5) probe used in the bDNA technology comprises target-probe (target probe), front amplification body (preamplifier), amplifies body (amplifier), label probe (label probe), in above-mentioned probe, only have target-probe to carry out different designs for different goal gene, rear three kinds of probes are all general to the bDNA analysis of any gene, target-probe has comprised the multiple specific probe for PCA3 gene different fragments, for the captured target gene fragment, in order to draw result more accurately.
The invention has the beneficial effects as follows, utilize the bDNA technology to be good at the catching of small segment gene, the efficient specific combination of the target-probe of unique design and PCA3, add that improved amplifying signal system makes sensitivity, specificity all guaranteed.This technology has been avoided false positive, the Problem of False Negative of normal PCR, and simple to operate, and all have molecular biology experiment basic skills personnel and all can operate.Simultaneously, the result that present technique detects is highly sensitive, positive rate is high, stability and favorable repeatability, easy and simple to handle, save time, be easy to popularize.
Embodiment
In order to make purpose of the present invention, technical scheme and advantage clearer, below in conjunction with embodiment, the present invention is further elaborated.Should be appreciated that specific embodiment described herein, only in order to explain the present invention, is not intended to limit the present invention.
A kind of BDNA detects the application of PCA3 in urine, and the operation steps adopted is:
(1) the PCA3 sample is collected
Urine PCA3 detects the urine specimen after using sample for the per rectum massage of prostate:
1) check that front drinking-water 500ml contributes to collect urine specimen;
2) prostate gland refers to collect immediately urine after inspection;
3) at least wanting the 20-30ml urine, be collected in the cup that posts label, must be the urine specimen of initial excretion, in the urine cup of preservation urine, should not contain any sanitas;
4), if the sample prolonged preservation should be put-80 ℃, detect in a short time and place-20 ℃ of preservations.
(2) research PCA3 sample is collected
1) research and utilization PCA3 as target the feasibility in prostate cancer diagnosis individuation assessment;
2) research bDNA technology platform detects after the per rectum massage of prostate sensitivity, specificity, the detection speed of PCA3 scoring diagnosing prostate cancer in urine, and whether is convenient to flux and detects;
3) the patients with prostate cancer PCA3 scoring of different clinical stagess of research and pathological grading changes whether significant difference and clinical meaning are arranged;
4) research PCA3 scoring, as the feasibility of monitoring index after prostate cancer progress and prognosis judging quota and prostate cancer therapy;
5) the ROC curve of Study of China people PCA3 and standards of grading and diagnostic threshold;
6) research PCA3 isomer is in Chinese patients with prostate cancer crowd's expression and whether variant expression in the tumor specimen of different pathological classification;
7) under research PCA3 different isomerization body surface reaches, androgen receptor isomer expression, whether analyze both has dependency.
(3) utilize the BDNA technology for detection:
1) set up the optimum detection flow process with PCA3 early diagnosis prostate gland cancer in bDNA technology for detection urine;
2) set up ROC curve and the PCA3 diagnostic threshold of Chinese PCA3 scoring, and PCA3 is to the evaluation system in the prostate cancer Individual Diagnosis; And the mark using PCA3 as the Noninvasive diagnosis prostate cancer is at clinical application;
3) understand Chinese patients with prostate cancer PCA3 isomer expression specificity;
4) understand Chinese patients with prostate cancer PCA3 isomer and express the dependency of expressing with the androgen receptor isomer.
5) probe used in the bDNA technology comprises target-probe (target probe), front amplification body (preamplifier), amplifies body (amplifier), label probe (label probe), in above-mentioned probe, only have target-probe to carry out different designs for different goal gene, rear three kinds of probes are all general to the bDNA analysis of any gene, target-probe has comprised the multiple specific probe for PCA3 gene different fragments, for the captured target gene fragment, in order to draw result more accurately.
The experimental data drawn: PCA3 probe design preliminary identification
The PCA3 probe data is analyzed, consistence initial survey, 32 samples
PSA probe design preliminary identification
The positive and probable positive sample check result contrast with pathology
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any modifications of doing within the spirit and principles in the present invention, be equal to and replace and improvement etc., within all should being included in protection scope of the present invention.
Claims (1)
1. a BDNA detects the application of PCA3 in urine, it is characterized in that, the operation steps adopted is:
(1) the PCA3 sample is collected
Urine PCA3 detects the urine specimen after using sample for the per rectum massage of prostate:
1) check that front drinking-water 500ml contributes to collect urine specimen;
2) prostate gland refers to collect immediately urine after inspection;
3) at least wanting the 20-30ml urine, be collected in the cup that posts label, must be the urine specimen of initial excretion, in the urine cup of preservation urine, should not contain any sanitas;
4), if the sample prolonged preservation should be put-80 ℃, detect in a short time and place-20 ℃ of preservations.
(2) research PCA3 sample is collected
1) research and utilization PCA3 as target the feasibility in prostate cancer diagnosis individuation assessment;
2) research bDNA technology platform detects after the per rectum massage of prostate sensitivity, specificity, the detection speed of PCA3 scoring diagnosing prostate cancer in urine, and whether is convenient to flux and detects;
3) the patients with prostate cancer PCA3 scoring of different clinical stagess of research and pathological grading changes whether significant difference and clinical meaning are arranged;
4) research PCA3 scoring, as the feasibility of monitoring index after prostate cancer progress and prognosis judging quota and prostate cancer therapy;
5) the ROC curve of Study of China people PCA3 and standards of grading and diagnostic threshold;
6) research PCA3 isomer is in Chinese patients with prostate cancer crowd's expression and whether variant expression in the tumor specimen of different pathological classification;
7) under research PCA3 different isomerization body surface reaches, androgen receptor isomer expression, whether analyze both has dependency.
(3) utilize the BDNA technology for detection:
1) set up the optimum detection flow process with PCA3 early diagnosis prostate gland cancer in bDNA technology for detection urine;
2) set up ROC curve and the PCA3 diagnostic threshold of Chinese PCA3 scoring, and PCA3 is to the evaluation system in the prostate cancer Individual Diagnosis; And the mark using PCA3 as the Noninvasive diagnosis prostate cancer is at clinical application;
3) understand Chinese patients with prostate cancer PCA3 isomer expression specificity;
4) understand Chinese patients with prostate cancer PCA3 isomer and express the dependency of expressing with the androgen receptor isomer.
5) probe used in the bDNA technology comprises target-probe (target probe), front amplification body (preamplifier), amplifies body (amplifier), label probe (label probe), in above-mentioned probe, only have target-probe to carry out different designs for different goal gene, rear three kinds of probes are all general to the bDNA analysis of any gene, target-probe has comprised the multiple specific probe for PCA3 gene different fragments, for the captured target gene fragment, in order to draw result more accurately.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012103914886A CN103436594A (en) | 2012-10-16 | 2012-10-16 | Application of BDNA for detecting PCA3 in urine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012103914886A CN103436594A (en) | 2012-10-16 | 2012-10-16 | Application of BDNA for detecting PCA3 in urine |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103436594A true CN103436594A (en) | 2013-12-11 |
Family
ID=49690308
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012103914886A Pending CN103436594A (en) | 2012-10-16 | 2012-10-16 | Application of BDNA for detecting PCA3 in urine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103436594A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106701984A (en) * | 2017-02-07 | 2017-05-24 | 华南师范大学 | Electrochemical luminescence nucleic acid detection method and kit based on branched DNA (Deoxyribonucleic Acid) amplification signal |
| CN108504738A (en) * | 2018-04-27 | 2018-09-07 | 郑州科蒂亚生物技术有限公司 | A kind of prostate cancer detection probe sequence and its application |
| CN109762905A (en) * | 2019-03-11 | 2019-05-17 | 天津脉络医学检验有限公司 | It is a kind of for assist detection prostate cancer ddPCR Primer composition and its application |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999049083A1 (en) * | 1998-03-24 | 1999-09-30 | Urocor, Inc. | Diagnosis of disease state using mrna profiles in peripheral leukocytes |
| WO2002012440A2 (en) * | 2000-08-07 | 2002-02-14 | Gene Logic, Inc. | Identifying drugs for and diagnosis of benign prostatic hyperplasia using gene expression profiles |
| CN101423873A (en) * | 2008-11-26 | 2009-05-06 | 苏州大学 | Kit for detecting human prostate cancer specific gene DD3<PCA3> by loop-mediated isothermal amplification |
| CN101652484A (en) * | 2006-11-08 | 2010-02-17 | 密歇根大学董事会 | Prostate cancer marker SPINK1 and application thereof |
| CN101942523A (en) * | 2010-10-15 | 2011-01-12 | 邵棠 | Liquid-phase chip method for detecting PCA3 gene and PSA gene and diagnostic reagent kit thereof |
-
2012
- 2012-10-16 CN CN2012103914886A patent/CN103436594A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999049083A1 (en) * | 1998-03-24 | 1999-09-30 | Urocor, Inc. | Diagnosis of disease state using mrna profiles in peripheral leukocytes |
| WO2002012440A2 (en) * | 2000-08-07 | 2002-02-14 | Gene Logic, Inc. | Identifying drugs for and diagnosis of benign prostatic hyperplasia using gene expression profiles |
| CN101652484A (en) * | 2006-11-08 | 2010-02-17 | 密歇根大学董事会 | Prostate cancer marker SPINK1 and application thereof |
| CN101423873A (en) * | 2008-11-26 | 2009-05-06 | 苏州大学 | Kit for detecting human prostate cancer specific gene DD3<PCA3> by loop-mediated isothermal amplification |
| CN101942523A (en) * | 2010-10-15 | 2011-01-12 | 邵棠 | Liquid-phase chip method for detecting PCA3 gene and PSA gene and diagnostic reagent kit thereof |
Non-Patent Citations (4)
| Title |
|---|
| DAPHNE HESSELSA ET AL.: "DD3PCA3-based Molecular Urine Analysis for the Diagnosis of Prostate Cancer", 《EUROPEAN UROLOGY》, vol. 44, 31 December 2003 (2003-12-31), pages 8 - 16, XP009032062, DOI: doi:10.1016/S0302-2838(03)00201-X * |
| 刘光香等: "PCA3 在前列腺癌患者前列腺按摩后尿液中的表达", 《现代泌尿外科杂志》, vol. 14, no. 1, 31 January 2009 (2009-01-31), pages 18 - 21 * |
| 刘龙亚: "外周血、尿PCA3和PSA基因的表达及尿PCA3评分在前列腺癌诊断中的意义", 《苏州大学硕士学位论文》, 31 May 2011 (2011-05-31), pages 1 - 58 * |
| 戴美洁等: "尿液中PCA3mRNA和融合基因TMPRSS2：ERG mRNA的检测在前列腺癌早期临床诊断中的应用", 《医学研究杂志》, vol. 40, no. 9, 30 September 2011 (2011-09-30), pages 20 - 24 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106701984A (en) * | 2017-02-07 | 2017-05-24 | 华南师范大学 | Electrochemical luminescence nucleic acid detection method and kit based on branched DNA (Deoxyribonucleic Acid) amplification signal |
| CN108504738A (en) * | 2018-04-27 | 2018-09-07 | 郑州科蒂亚生物技术有限公司 | A kind of prostate cancer detection probe sequence and its application |
| CN108504738B (en) * | 2018-04-27 | 2021-09-10 | 郑州科蒂亚生物技术有限公司 | Prostate cancer detection probe sequence and application thereof |
| CN109762905A (en) * | 2019-03-11 | 2019-05-17 | 天津脉络医学检验有限公司 | It is a kind of for assist detection prostate cancer ddPCR Primer composition and its application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Sanguedolce et al. | Urine TMPRSS2: ERG fusion transcript as a biomarker for prostate cancer: literature review | |
| CN106459961B (en) | Pancreatic cancer detection kit or device and detection method | |
| Day et al. | PCA3: from basic molecular science to the clinical lab | |
| CN106414774B (en) | Detection kit or device for biliary tract cancer and detection method | |
| Allan et al. | Circulating tumor cell analysis: technical and statistical considerations for application to the clinic | |
| Maheswaran et al. | Circulating tumor cells: a window into cancer biology and metastasis | |
| CN106459964B (en) | Esophageal cancer detection kit or device and detection method | |
| ES2511218T5 (en) | Kits and products for detection of nucleic acids in individual cells and for identification of rare cells in large populations of heterogeneous cells | |
| JP2008533487A (en) | Methods to predict progression-free and overall survival at each follow-up period point during therapy for patients with metastatic breast cancer using circulating tumor cells | |
| CN104004840B (en) | Test kit for early screening Yu diagnosis of prostate cancer | |
| US8512949B2 (en) | Diagnosis/treatment option for head-and-neck tumor using micro-RNA as biomarker | |
| CN101942523B (en) | Liquid-phase chip method for detecting PCA3 gene and PSA gene and diagnostic reagent kit thereof | |
| CN106636390A (en) | Gene methylation detection method, detection kit and application thereof | |
| KR20120132592A (en) | Method of providing information for diagnosis of cancer using quantitative realtime PCR and diagnostic kit comprising thereof | |
| CN104498495A (en) | Prostatic cancer marker lncRNA MALAT-1 and application thereof | |
| CN103436594A (en) | Application of BDNA for detecting PCA3 in urine | |
| JP6611411B2 (en) | Pancreatic cancer detection kit and detection method | |
| CN104531863A (en) | PCA3 gene detection method and primer in human urine | |
| CN104164515B (en) | A kind of Honghu myxobolus specific PCR detection primer and detection method thereof | |
| CN106119361A (en) | The test kit detected for the NDRG4 gene methylation of early gastric caacer diagnosis and prognosis evaluation and application thereof | |
| US20170073766A1 (en) | Method for differentiating between lung squamous cell carcinoma and lung adenocarcinoma | |
| CN108949923A (en) | A kind of method and kit and application expanding MSF1 gene | |
| CN102732637B (en) | Multiplex nested methylation specific PCR (Polymerase Chain Reaction) detection kit, using method and application thereof | |
| CN107326092A (en) | Applications and colorectal cancer detection kit of the related miRNA of colorectal cancer as biomarker | |
| JP2018057359A (en) | Method of predicting effect of anticancer drug administration |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20131211 |