EP0904397A4 - Methode zur erkennung des prostat-spezifischen antigens zur überwachung und diagnose von prostatkrebs - Google Patents

Methode zur erkennung des prostat-spezifischen antigens zur überwachung und diagnose von prostatkrebs

Info

Publication number
EP0904397A4
EP0904397A4 EP97918724A EP97918724A EP0904397A4 EP 0904397 A4 EP0904397 A4 EP 0904397A4 EP 97918724 A EP97918724 A EP 97918724A EP 97918724 A EP97918724 A EP 97918724A EP 0904397 A4 EP0904397 A4 EP 0904397A4
Authority
EP
European Patent Office
Prior art keywords
seq
prostate cancer
psa
prostate
monitoring
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.)
Withdrawn
Application number
EP97918724A
Other languages
English (en)
French (fr)
Other versions
EP0904397A1 (de
Inventor
David J Robbins
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
SmithKline Beecham Corp
Original Assignee
SmithKline Beecham Corp
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by SmithKline Beecham Corp filed Critical SmithKline Beecham Corp
Publication of EP0904397A1 publication Critical patent/EP0904397A1/de
Publication of EP0904397A4 publication Critical patent/EP0904397A4/de
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6424—Serine endopeptidases (3.4.21)
    • C12N9/6445—Kallikreins (3.4.21.34; 3.4.21.35)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/112—Disease subtyping, staging or classification

Definitions

  • RT-PCR reverse-transcriptase PCR
  • cDNA complementary DNA
  • RT-PCR can thus reveal by amplification the presence of a single species of mRNA. Accordingly, if the mRNA is highly specific for the cell that produces it, RT-PCR can be used to identify the presence of a specific type of cell.
  • PSA Prostate-specific antigen
  • RT-PCR of mRNA encoding for PSA has been used in the identification of prostate cancer cells.
  • Deguchi et al. detected PSA mRNA in regional lymph nodes from patients with prostate cancer which had metastasized to the lymphatic system. Cancer Res. 1993, 53:5350-4.
  • RT-PCR methods for detection of prostate cancer cells in the general circulation of patients with advanced prostate cancer have also been disclosed. Moreno et al. Cancer Res. 1992, 53:6110-2; Jaakkola et al. Clin. Chem. 1995, 41(2): 182- 186.
  • patients with early prostate cancer and/or well differentiated or localized disease were found negative for PSA mRNA by PCR. Accordingly, it was concluded that RT-PCR for PSA mRNA would not be a useful tool for either the diagnosis or monitoring of prostate cancer. Diamandis, E.P. and Yu, H. Clin. Chem. 1995, 41(2): 177-179.
  • the present invention provides a new RT-PCR method with novel primers for the monitoring and diagnosis patients with prostate cancer.
  • This method is designed to determine the presence of mRNA for PSA in the bloodstream thereby providing evidence that prostate cells are circulating in the blood and confirming a diagnosis of cancer. Circulating prostate cells also indicate that the patient is at risk of secondary tumor or bone metastatic anchoring. Samples taken after prostatectomy or radiation treatment which are reactive indicate the risk that the prostate cancer had spread and that surgery/radiation was not efficacious.
  • An object of the present invention is to provide primers for use in an RT-PCR method for the detection of prostatic specific antigen for the monitoring and diagnosis of patients with prostate cancer.
  • Figure 1 provides the human DNA for prostate specific antigen (PSA) (SEQ ID NOS: 1, 8, 9 and 10). Introns are abbreviated for clarity. Exons are indicated in bold. Primers previously disclosed in the art are indicated by underlining. The primers of the present invention, namely DR PSA-up (SEQ ED NO: 2), DR PSA- Down (SEQ ID NO: 3) and DR 12 mer (SEQ ID NO: 4), are indicated by underlined italics in a larger font size.
  • PSA prostate specific antigen
  • a new method for the detection of prostate specific antigen by reverse- transcriptase polymerase chain reaction has been developed which can be used in the monitoring of prostate cancer in a patient.
  • Primers which are used in the method of the present invention include DR PSA-Up: 5'-GTTGTCITCCTCACCCrGTCCG-3' (SEQ LO NO: 2), DR PSA-Down: S'-TCCAGCACACAGCATGAACTTG-S' (SEQ LD NO: 3), and DR 12 mer: 5'-GAATCACCCGAG-3' (SEQ LD NO: 4).
  • the method of the present invention comprises isolation and washing of buffy coat cells, isolation of total RNA from buffy coat cells, reverse transcription (RT) reaction for PSA, PCR for PSA, and detection of the absence or presence of PSA product on a DNA sequencer.
  • the method may further comprise RT reaction and PCR for a housekeeping gene Beta-2-microglobulin (B2G: Israeli et al. Cancer Research 1994 54:6306-6310) to momtor quality of the isolated RNA.
  • B2G Israeli et al. Cancer Research 1994 54:6306-6310
  • Vacutainer Systems Cat. #362761 Sodium Citrate anticoagulant, 8 ml draw, San Jose, CA are used to collect blood samples from patients suspected of having prostate cancer. These samples are then centrifuged in accordance with the manufacturer's instructions to separate the buffy coat from other blood components. Buffy coat cells are then washed with PBS and pelleted by centrifugation. The pelleted cells are taken up in a cell lysis buffer. Cells are homogenized by passage over a Qiagen QIA shredder microspin column. The run-through is then passed over a Qiagen RNeasy column to bind RNA. The sample is washed to remove DNA and other components, and the RNA is eluted with DEPC-treated water.
  • the eluted sample RNA is then used as the substrate in the RT reactions.
  • the RT reaction primer for PSA reactions is DR 12 mer (SEQ ID NO: 4) which is specific for the PSA target.
  • An aliquot of the RT reaction is then used for the PCR reaction for PSA, using primers DR PSA Up (SEQ ID NO: 2) and DR PSA Down (SEQ ID NO: 3).
  • the method further comprises RT reaction and PCR for B2G to monitor quality of the isolated RNA
  • the RT reaction primer is 5'-AGCTTTGAGTGC-3' (SEQ ED NO: 5) and the PCR reaction primers are 5'-AGCAGAGAATGGAAAGTCAAA-3' (SEQ ED NO: 6) and 5 -TGT TGATGT TGGATAAGAGAAT-3 1 (SEQ ID NO: 7).
  • the sizes of the products from each reaction for each patient are compared on the ABI 373 DNA Analyzer with the ABI 672 GENESCAN software.
  • LNCaP prostate cancer tissue culture
  • WBCs white blood cells
  • the ABI 672 GENESCAN software can determine the size in bases of each product, allowing definitive identification of each product.
  • BPH samples included patients diagnosed with BPH but not cancer. Prostate cancer status was provided by the patient's physician.
  • the method of the present invention is specific for determining metastatic prostate cancer, in particular prostate cancer in the progressive stage and that which has been stable for less than 5 years.
  • progressive it is meant cancer with identified metastases to the bone or other sites, with the metastases enlarged and/or more numerous than in the last examination.
  • the results in Table 1 are consistent with this relapse rate. Tests with the assay of the present invention predict that the 2 out of 6 nonreactive patients in this group will remain stable for over 5 years while the 4 out of 6 reactive patients have a high probability of relapse.
  • Samples from patients having stage B cancer, or organ-confined cancer, which is not metastatic were nonreactive in the method of the present invention. Further, stable patients, meaning those with metastases in the past but no further progression in 5 or more years, who are considered cured, were also nonreactive. Recent studies indicate that circulating prostate cells can also result from prostatectomy operations. Oefelein et al. J. Urol. 1996 155:238-242. If cancerous, these circulating cells caused by the surgery could result in metastases. The method of the present invention can be used in monitoring these surgical procedures to ensure that a patient is nonreactive prior to surgery as well as after the surgery.
  • kits for monitoring of progression of prostate cancer which comprise the DR PSA-Up (SEQ ED NO: 2), DR PSA-Down (SEQ ID NO: 3), and DR 12 mer (SEQ ID NO: 4) primers.
  • these kits also comprise primers for RT and PCR detection of B2G such as SEQ ED NOs: 5, 6 and 7.
  • kits of the present invention may comprise tubes for collection of blood samples which are capable of separating the buffy coat from other blood components such as Vacutainer Cell Preparation Tubes.
  • Kits of the present invention may also comprise a means for isolating RNA.
  • the kit may comprise a cell lysis buffer, Qiagen QIA shredder microspin columns (Cat. #79655) for cell homogenization and Qiagen RNeasy columns (Cat. #74106) for binding of the RNA.
  • the kits may also contain any of the following reagents including, but not limited to, Beta-Mercaptoethanol (BME), Diethyl Pyrocarbonate (DEPC), Superscript II RNase H- Reverse Transcriptase (Gibco BRL, Gaithersburg, MD), ethanol (200 Proof), 100 mM dNTPs, AmpliTaq DNA Polymerase (Perkin Elmer Cat. #N808-0153), 6% (6% T, 5%C) denaturing acrylamide gel Gibco/BRL 6% Sequencing Solution (Gel-Mix 6; Cat. #5543UA), 10 % ammonium persulfate,
  • Denaturing loading buffer containing Blue dextran (50 mg/ml) and 200 ml formamide and 20 ml 100 mM EDTA, IX TBE for Sequencer Gibco/BRL UltraPure Gel-Mix running mate (Cat. #15546-013) and Brij 35 (Sigma Chemical Co. St. Louis, MO, Cat. #430AG-6). Standard and controls may also be provided in the kits of the present invention. Examples include, but are not limited to Genescan ROX-2500 standard (Perkin Elmer/ABI Cat.
  • RNA from ATCC cell lines CRL-1435 PC-3 prostate adenocarcinoma, human
  • HTB-22 MCF7 breast adenocarcinoma, pleural effusion, human
  • RNA from ATCC cell line CRL-1740 LNCaP metalastatic prostate adenocarcinoma, human
  • Reagents are prepared in essentially RNAse-free, sterile, disposable plasticware, in glassware baked at 180°C for at least 8 hours, or in polypropylene plasticware rinsed with chloroform.
  • Solutions used for RNA and reverse transcription work are prepared using RNAse-free glassware, autoclaved water, and chemicals reserved for RNA work handled with baked spatulas. Water for solutions is treated with 0.1% DEPC for at least 12 hours at 37°C and then heated to 100°C for 15 minutes or autoclaved for 15 minutes at 15 lb/sq. inch on liquid cycle.
  • Phosphate buffered saline (PBS) used in this method is prepared by dissolving 8 grams of NaCl, 0.2 grams of KCl, 1.44 grams of Na.HPO 4 , and 0.24 grams of KH-PO 4 in 800 ml of distilled ILG The pH of the buffer is adjusted to 7.4 with HCl. The volume is then adjusted to 1 liter with H.O. Prior to use, the buffer is sterilized by autoclaving for 20 minutes at 15 lb/sq. inch on liquid cycle and stored at room temperature. Lysis Buffer (BME) is prepared from 100 ⁇ l of BME per 10 ml of QIAGEN
  • the resulting Lysis Buffer/BME is stable for 1 month.
  • Wash Buffer RPE is prepared for use by adding 4 volumes of 96-100% ethanol to the concentrate supplied by Qiagen.
  • Seventy percent Ethanol is prepared by adding 700 ml of 200 proof Ethanol to 300 ml DEPC-treated water. The solution is stored at room temperature.
  • Blood samples are obtained in Vacutainer CPT tubes with sodium citrate. The samples are then centrifuged at room temperature (20 - 25°C) in a horizontal swing-out head rotor at 1500 to 1800 RCF (Relative Centrifugal Force) for 30 minutes. After centrifugation, the buffy coat appears as a whitish layer just under the plasma layer. Approximately half of the plasma is aspirated without disturbing the cell layer. The cell layer is then collected with a sterile pipette and transferred to a 15 ml conical centrifuge tube.
  • RCF Relative Centrifugal Force
  • PBS is added to the conical tubes to bring the volume to 15 ml.
  • the tubes are then capped and the cells and PBS mixed by inverting the tubes 5 times.
  • the tubes are then centrifuged for 15 minutes at 300 RCF and the supernatant is aspirated without disturbing the cell pellet.
  • PBS is again added to the cells to bring the volume to 10 ml.
  • the tubes are capped and the cells and PBS are mixed by inverting the tubes 5 times.
  • the tubes are then centrifuged for 10 minutes at 300 RCF. As much supernatant as possible is aspirated without disturbing the cell pellet.
  • Lysis Buffer/BME 400 ⁇ i
  • samples may be frozen at -70°C or used immediately for RNA isolation.
  • Example 3 RNA Isolation The cells/Lysis Buffer/BME suspension is pipetted directly onto a QIAGEN
  • Wash Buffer RPE 500 ⁇ l is pipetted onto the spin column and centrifuge as above. The flowthrough is discarded. A second aliquot of Wash Buffer RPE is pipetted onto the spin column and centrifuged for 2 minutes at full speed to dry the RNeasy spin column. This 2 minute spin assures that no residual ethanol will be carried over during elution. The spin column is then transferred to a new 1.5 ml collection tube and the sample RNA is eluted with 50 ⁇ l of DEPC-treated water pipetted directly onto the spin column membrane followed by centrifugation for 60 seconds at 8000 x g. Sample RNA may be used immediately in the RT reaction, or stored at -70°C until further use.
  • Example 4 Determining the RNA concentration
  • RNA concentration in each sample is determined by UV spectrophotometry.
  • the baseline absorbance of 75 ⁇ l of distilled water is first determined at 260, 280, and 320 nm.
  • a five microliter sample of RNA is then added and mixed.
  • the sample absorbance is then determined at 260, 280, and 320 nm.
  • the neat RNA concentration is calculated by: a) subtracting the A320 from the A260 to obtain the absorbance due to RNA; b) multiplying by the dilution factor (80 ⁇ /5 ⁇ l); and c) multiplying by the conversion factor (0.04 mg/ ml).
  • Example 5 Sample Reverse Transcription
  • Two microliters of 50 ⁇ M PSA RT primer (SEQ ED NO: 4) are pipetted into each PCR reaction tubes.
  • Two microliters of 50 ⁇ M B2G RT primer (SEQ ID NO: 5) are pipetted into separate PCR reaction tubes. Tubes may be prepared in advance and stored at -20°C. RT Reaction Mix is prepared just prior to use in the following manner: The number of sample reactions (PSA and B2G) to be run is first determined as variable X.
  • PSA PCR Reaction Mix is prepared in the following manner.
  • the number of PSA sample reactions to be run is determined as variable X.
  • the following volumes of reagents are then mixed: 31.95(X+1) ⁇ l H ⁇ O; 4(X+1) ⁇ l 10X PCR Buffer 0; (X+l) ⁇ l 20 mM dNTP; 0.8(X+1) ⁇ l 25 mM MgCl 2 ; 2(X+1) ⁇ l PSA primers (SEQ ID NOs: 2 and 3); and 0.25(X+1) ⁇ l Taq enzyme.
  • B2G PCR Reaction Mix is prepared in the following manner: The number of B2G sample reactions to be run is determined as variable X. The following volumes of reagents are then mixed: 33.15(X+1) ⁇ l H j O; 4(X+1) ⁇ l 10X PCR Buffer JJ; (X+l) ⁇ l 20 mM dNTP; 0.8(X+1) ⁇ l 25 mM MgCL; 0.8(X+1) ⁇ l B2G primers (SEQ ED NOs: 6 and 7); and 0.25(X+1) ⁇ l Taq enzyme.
  • RNA sample Two tubes per RNA sample are prepared, one tube for each reaction: PSA and B2G. Forty microliters of the appropriate reaction mixture are pipetted into each of the tubes. Ten microliters of the respective sample RNA or control RNA Reverse Transcription reaction are pipetted into each tube. For the negative (reaction) control, 10 ml of water are used. The tubes are spun for 10 seconds in an Eppendorf microcentrifuge to ensure all liquid is mixed together at the bottom of the tubes. The tubes are then placed in the Perkin Elmer 9600 Thermocycler and cycled as follows:
  • Example 7 Sample Detection Two 6 cm well-to-read plates (one notched, one plain) are prepared for use in the following manner. Both sides of each plate are rinsed with distilled water. One side only of each plate is then selected as the inside (gel side) of the plate. This inside is then rinsed with methanol followed by 0.5% Brij. The methanol rinse is then repeated until the plates are streak free. Spacers are then placed on the edge of the plain plate, parallel to the short side of the plates. The notched plate is placed on top of the plain plate, with the insides of the plates facing each other. The plates are then stood on their bottom edge and the spacers lined up so they are level with the bottom and sides of the gel. They are gently laid down flat with the plain plate on the bottom and clamped with 6 binder clips.
  • the clamps are removed and the outside of the plates are cleaned with distilled water, being careful to remove all acrylamide from the outside surfaces ofthe plates.
  • the plates are then inserted into a 373 sequencer and a plate check is performed in accordance with the manufacturer's directions to check for interference.
  • the top and bottom buffer reservoirs are filled with 700 ml and 300 ml of IX TBE, respectively.
  • the sample comb is removed from between the plates, and acrylamide and urea are washed from the wells with IX TBE.
  • the gel is prerun for 5 minutes at 28 watts.
  • Two microliters of PCR reaction are pipetted into a 0.2 ml thin-walled PCR reaction tube.
  • Denaturing sample buffer (2X; 2.5 ml) and 0.3 ml of Genescan ROX-2500 standard are then added.
  • the tubes are placed in a PE 9600 Thermocycler and run at 94 °C for 10 minutes.
  • the thermal cycler is set to PAUSE and each tube is placed on ice.
  • Four microliters of cooled sample are then loaded onto the gel.
  • the ABI sequencer is set to run for 3.5 hours at 28 watts, 600 volts, 40 milliamps and the Genescan data collection to collect for 3 hours.
  • TELECOMMUNICATION INFORMATION (A) TELEPHONE: 610-270-5219 (B) TELEFAX: 610-270-5090
  • GACTCCAGCC ACGACCTCAT GCTGCTCCGC CTGTCAGAGC CTGCCGAGCT CACGGATGCT 240

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Microbiology (AREA)
  • Analytical Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Pathology (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Biochemistry (AREA)
  • Oncology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Hospice & Palliative Care (AREA)
  • Medicinal Chemistry (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
EP97918724A 1996-04-16 1997-04-16 Methode zur erkennung des prostat-spezifischen antigens zur überwachung und diagnose von prostatkrebs Withdrawn EP0904397A4 (de)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US1576596P 1996-04-16 1996-04-16
US15765P 1996-04-16
PCT/US1997/006497 WO1997039139A1 (en) 1996-04-16 1997-04-16 A method for detection of prostate specific antigen used in monitoring and diagnosis of prostate cancer

Publications (2)

Publication Number Publication Date
EP0904397A1 EP0904397A1 (de) 1999-03-31
EP0904397A4 true EP0904397A4 (de) 2003-01-29

Family

ID=21773475

Family Applications (1)

Application Number Title Priority Date Filing Date
EP97918724A Withdrawn EP0904397A4 (de) 1996-04-16 1997-04-16 Methode zur erkennung des prostat-spezifischen antigens zur überwachung und diagnose von prostatkrebs

Country Status (5)

Country Link
EP (1) EP0904397A4 (de)
JP (1) JP2000510331A (de)
AU (1) AU2676197A (de)
WO (1) WO1997039139A1 (de)
ZA (1) ZA973231B (de)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2006201736B2 (en) * 1998-08-10 2008-08-28 Agensys, Inc. BPC-1: a secreted brain specific protein expressed and secreted by prostate and bladder cancer cells
DE69934694T2 (de) 1998-08-10 2007-10-04 Agensys, Inc., Santa Monica Bpc-1: ein ausgeschiedenes gehirnspezifisches protein das von prostata- und blasen-krebszellen exprimiert und ausgeschieden wird
EP1151142A2 (de) 1999-01-28 2001-11-07 Gen-Probe Incorporated Nukleinsäure sequenzen für den nachweis von genetischen markern für krebs in einer biologischen probe
WO2004073657A2 (en) * 2003-02-19 2004-09-02 Protein Design Labs, Inc. Methods of diagnosis of cancer and other diseases, composition and methods of screening for modulators of cancer and other diseases

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0520794A1 (de) * 1991-06-26 1992-12-30 F. Hoffmann-La Roche Ag Methoden zur Feststellung von Krebsmetastasen durch Nukleinsäure-Amplifizierung

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2148350A1 (en) * 1992-10-29 1994-05-11 Carlo Croce Methods of detecting micrometastasis of prostate cancer

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0520794A1 (de) * 1991-06-26 1992-12-30 F. Hoffmann-La Roche Ag Methoden zur Feststellung von Krebsmetastasen durch Nukleinsäure-Amplifizierung

Non-Patent Citations (9)

* Cited by examiner, † Cited by third party
Title
DEGUCHI T ET AL: "DETECTION OF MICROMETASTATIC PROSTATE CANCER CELLS IN LYMPH NODES BY TRANSCRIPTASE-POLYMERASE CHAIN REACTION", CANCER RESEARCH, AMERICAN ASSOCIATION FOR CANCER RESEARCH, BALTIMORE, MD, US, vol. 53, 15 November 1993 (1993-11-15), pages 5350 - 5354, XP002059297, ISSN: 0008-5472 *
DIGBY M ET AL: "HUMAN PROSTATE SPECIFIC ANTIGEN (PSA) GENE: STRUCTURE AND LINKAGE TO THE KALLIKREIN-LIKE GENE, HGK-1", NUCLEIC ACIDS RESEARCH, OXFORD UNIVERSITY PRESS, SURREY, GB, vol. 17, no. 5, 1989, pages 2137, XP000999289, ISSN: 0305-1048 *
PANTEL K AND RIETHMÜLLER G: "Methods for detection of micrometastatic carcinoma cells in bone marrow, blood, and lymph nodes", ONKOLOGIE, KARGER, FREIBURG, DE, vol. 18, 1995, pages 394 - 401, XP002092612, ISSN: 0378-584X *
PELKEY J P ET AL: "Molecular and immunological detection of circulating tumor cells and micrometastases from solid tumors", CLINICAL CHEMISTRY, AMERICAN ASSOCIATION FOR CLINICAL CHEMISTRY. WINSTON, US, vol. 42, no. 9, 1996, pages 1369 - 1381, XP002092613, ISSN: 0009-9147 *
RIEGMAN P H J ET AL: "Characterization of the prostate-specific gene: A novel human Kallikrein-like gene", BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, ACADEMIC PRESS INC. ORLANDO, FL, US, vol. 159, no. 1, 28 February 1989 (1989-02-28), pages 95 - 102, XP002141957, ISSN: 0006-291X *
See also references of WO9739139A1 *
STONE N ET AL: "DETECTION OF CIRCULATING METASTATIC TUMOR CELLS BY PSA AND PSM MAY BE USEFUL TO IDENTIFY SUBGROUPS OF ADVANCED PROSTATE CANCER PATIENTS", CANCER RESEARCH, AMERICAN ASSOCIATION FOR CANCER RESEARCH, BALTIMORE, MD, US, vol. 37, 1 March 1996 (1996-03-01), pages 247, XP002910105, ISSN: 0008-5472 *
STRAY J ET AL: "REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION (RT-PCR) DETECTS METASTATIC PROSTATE CANCER CELLS IN LYMPH NODES, BLOOD AND POTENTIALLY IN BONE MARROW USING PSA-MRNA AS TEMPLATE", JOURNAL OF UROLOGY, BALTIMORE, MD, US, vol. 151, no. SUPPL 5, 1994, pages 412A, XP002910106, ISSN: 0022-5347 *
TAKAYAMA T K ET AL: "NEWER APPLICATIONS OF SERUM PROSTATE-SPECIFIC ANTIGEN IN THE MANAGEMENT OF PROSTATE CANCER", SEMINARS IN ONCOLOGY, BETHESDA, MD, US, vol. 21, no. 5, 1 October 1994 (1994-10-01), pages 542 - 553, XP002059296 *

Also Published As

Publication number Publication date
WO1997039139A1 (en) 1997-10-23
EP0904397A1 (de) 1999-03-31
AU2676197A (en) 1997-11-07
ZA973231B (en) 1997-11-25
JP2000510331A (ja) 2000-08-15

Similar Documents

Publication Publication Date Title
JP7358434B2 (ja) 尿バイオマーカーコホート、遺伝子発現特性、およびその使用の方法
Gomella et al. Reverse transcriptase polymerase chain reaction for prostate specific antigen in the management of prostate cancer
EP0667920B1 (de) Methoden zur detektion von mikrometastasen bei prostata-krebs
CA2513780C (en) Method to detect prostate cancer from a urine sample
US20090035773A1 (en) Nucleic acid sequences for detecting genetic markers for cancer in a biological sample
JP4944041B2 (ja) 前立腺癌の予後判定用マーカー及び／又はテラノスティックマーカーとなる、尿沈渣及び／又は尿のｍＲＮＡ比
US20090226925A1 (en) Methods for Detecting Circulating Tumor Cells
AU742753C (en) Primers and methods for the detection of disseminated tumor cells
WO1994010343A9 (en) Methods of detecting micrometastasis of prostate cancer
WO1998046788A9 (en) Primers and methods for the detection of disseminated tumor cells
US20260049300A1 (en) Diagnosis of prostate cancer
EP0904397A1 (de) Methode zur erkennung des prostat-spezifischen antigens zur überwachung und diagnose von prostatkrebs
US20090035774A1 (en) Method for characterizing primary tumors
CA2251557A1 (en) A method for detection of prostate specific antigen used in monitoring and diagnosis of prostate cancer
Fornari et al. New evaluation of plasma DNA microsatellite analysis in patients with TCC of the urinary bladder

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 19981007

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): BE CH DE DK FR GB IT LI NL

A4 Supplementary search report drawn up and despatched

Effective date: 20021216

RIC1 Information provided on ipc code assigned before grant

Ipc: 7C 12Q 1/68 B

Ipc: 7C 07H 21/04 B

Ipc: 7C 12P 19/34 A

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20030301