Reagents and methods useful for detecting diseases of the prostate

Inactive Publication Date: 2005-05-19
BILLING MEDEL PATRICIA +11
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0013] The present invention further provides a method of detecting a target PS118 polynucleotide in a test sample suspected of containing target PS118 polynucleotides, which comprises (a) contacting the test sample with at least one PS118 oligonucleotide as a sense primer and at least one PS118 oligonucleotide as an anti-sense primer, and amplifying same to obtain a first stage reaction product; (b) contacting the first stage reaction product with at least one other PS118 oligonucleotide to obtain a second stage reaction product, with the proviso that the other PS118 oligonucleotide is located 3′ to the PS118 oligonucleotides utilized in step (a) and is complementary to the first stage reaction product; and (c) detecting the second stage reaction product as an indication of the presence of a target PS118 polynucleotide in the test sample. The PS118 oligonucleotides selected as reagents in the method have at least 50% identity with a sequence selected from the group consisting of SEQUENCE ID NOS 1-10, and fragments or complements thereof. Amplification may be performed by the polymerase chain reaction. The test sample can be reacted either directly or indirectly with a solid phase prior to performing the method, or prior to amplification, or prior to detection. The detection step also comprises utilizing a detectable label capable of generating a measurable signal; further, the detectable label can be attached to a solid phase. Test kits useful for detecting target PS118 polynucleotides in a test sample are also provided which comprise a container containing at least one PS118-specific polynucleotide selected from the group consisting of SEQUENCE ID NOS 1-10, and fragments or complements thereof. These test kits further comprise containers with tools useful for collecting test samples (such as, for example, blood, urine, saliva and stool). Such tools include lancets and absorbent paper or cloth for collecting and stabilizing blood; swabs for collecting and stabilizing saliva; and cups for collecting and stabilizing urine or stool samples. Collection materials, such as papers, cloths, swabs, cups, and the like, may optionally be treated to avoid denaturation or irreversible adsorption of the sample. The collection materials also may be treated with or contain preservatives, stabilizers or antimicrobial agents to help maintain the integrity of the specimens.

Problems solved by technology

PSA, however, cannot differentiate between BPH and prostate cancer, which reduces its specificity as a marker for prostate cancer.
Further, if one could show that such a marker characteristic of aggressive cancer was absent, the patient could be spared expensive and non-beneficial treatment.

Method used

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  • Reagents and methods useful for detecting diseases of the prostate
  • Reagents and methods useful for detecting diseases of the prostate
  • Reagents and methods useful for detecting diseases of the prostate

Examples

Experimental program
Comparison scheme
Effect test

example 1

Identification of Prostate Tissue Library PS118 Gene-Specific Clones

[0198] A. Library Comparison of Expressed Sequence Tags (EST's) or Transcript Images. Partial sequences of cDNA clone inserts, so-called “expressed sequence tags” (EST's), were derived from cDNA libraries made from prostate tumor tissues, prostate non-tumor tissues and numerous other tissues, both tumor and non-tumor and entered into a database (LIFESEQ™ database, available from Incyte Pharmaceuticals, Palo Alto, Calif.) as gene transcript images. See International Publication No. WO 95 / 20681. (A transcript image is a listing of the number of EST's for each of the represented genes in a given tissue library. EST's sharing regions of mutual sequence overlap are classified into clusters. A cluster is assigned a clone number from a representative 5′ EST. Often, a cluster of interest can be extended by comparing its consensus sequence with sequences of other EST's which did not meet the criteria for automated clusterin...

example 2

Sequencing of PS118 EST-Specific Clones

[0200] The full-length DNA sequence of clone 717188 of the PS118 gene contig was determined (SEQUENCE ID NO 9) using dideoxy termination sequencing with dye terminators following known methods [F. Sanger et al., PNAS U.S.A. 74:5463 (1977)].

[0201] Because the pINCY vector (available from Incyte Pharmaceuticals, Inc., Palo Alto, Calif.) contains universal priming sites just adjacent to the 3′ and 5′ ligation junctions of the inserts, approximately 300 bases of the insert were sequenced in both directions using two universal primers (SEQUENCE ID NO 13 and SEQUENCE ID NO 14, available from New England Biolabs, Beverly, Mass., and Applied Biosystems Inc, Foster City, Calif., respectively). The sequencing reactions were run on a polyacrylamide denaturing gel, and the sequences were determined by an Applied Biosystems 377 Sequencer (available from Applied Biosystems, Foster City, Calif.). Additional sequencing primers (SEQUENCE ID NOS 15-22) were de...

example 3

Nucleic Acid

[0202] A. RNA Extraction from Tissue. Total RNA was isolated from prostate tissues and from non-prostate tissues. Various methods were utilized, including but not limited to the lithium chloride / urea technique, known in the art and described by Kato et al. (J. Virol. 61:2182-2191, 1987), and TRIzo™ (Gibco-BRL, Grand Island, N.Y.).

[0203] Briefly, tissue was placed in a sterile conical tube on ice and 10-15 volumes of 3 M LiCl, 6 M urea, 5 mM EDTA, 0.1 M β-mercaptoethanol, 50 mM Tris-HCl (pH 7.5) were added. The tissue was homogenized with a Polytron® homogenizer (Brinkman Instruments, Inc., Westbury, N.Y.) for 30-50 sec on ice. The solution was transferred to a 15 ml plastic centrifuge tube and placed overnight at −20° C. The tube was centrifuged for 90 min at 9,000×g at 04° C. and the supernatant was immediately decanted. Ten ml of 3 M LiCl were added and the tube was vortexed for 5 sec. The tube was centrifuged for 45 min at 11,000×g at 04° C. The decanting, resuspens...

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Abstract

A set of contiguous and partially overlapping cDNA sequences and polypeptides encoded thereby, designated as PS118 and transcribed from prostate tissue, is described. These sequences are useful for the detecting, diagnosing, staging, monitoring, prognosticating, preventing or treating, or determining the predisposition of an individual to diseases and conditions of the prostate, such as prostate cancer. Also provided are antibodies which specifically bind to PS118-encoded polypeptide or protein, and agonists or inhibitors which prevent action of the tissue-specific PS118 polypeptide, which molecules are useful for the therapeutic treatment of prostate diseases, tumors or metastases.

Description

CROSS-REFERENCE TO RELATED APPLICATION [0001] This application is a continuation-in-part of U.S. application Ser. No. 08 / 842,385 filed Apr. 23, 1997, from which priority is claimed pursuant to 35 U.S.C. §120 and which is incorporated herein by reference in its entirety.BACKGROUND OF THE INVENTION [0002] The invention relates generally to detecting diseases of the prostate, and more particularly, relates to reagents such as polynucleotide sequences and the polypeptide sequences encoded thereby, as well as methods which utilize these sequences, which methods are useful for detecting, diagnosing, staging, monitoring, prognosticating, preventing or treating, or determining predisposition to diseases or conditions of the prostate such as prostate cancer. [0003] Prostate cancer is the most common form of cancer occurring in males in the United States, with projections of 184,500 new cases diagnosed, and 39,200 related deaths to occur during 1998 (American Cancer Society). Prostate cancer ...

Claims

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Application Information

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IPC IPC(8): A61K38/00G01N33/53A61K39/395A61K45/00A61P13/08A61P35/00C07K14/47C07K16/18C12N1/15C12N1/19C12N1/21C12N5/10C12N15/09C12P21/02C12P21/08C12Q1/68G01N33/574
CPCC07K16/3069C12Q1/6886C12Q2600/136C12Q2600/158C12Q2565/518C12Q2531/113A61P13/08A61P35/00
Inventor BILLING-MEDEL, PATRICIACOHEN, MAURICECOLPITTS, TRACEYFRIEDMAN, PAULAGORDON, JULIANGRANADOS, EDWARDHODGES, STEVENKLASS, MICHAELKRATOCHVIL, JONROBERTS-RAPP, LISARUSSELL, JOHNSTROUPE, STEPHEN
Owner BILLING MEDEL PATRICIA
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