Method to detect prostate cancer in a sample
First Claim
1. A composition comprising:
- (a) a urine sample from a subject having or suspected of having prostate cancer, said urine sample comprising at least one prostate cell or nucleic acid extract thereof;
(b) a first oligonucleotide or first primer pair for performing an RNA hybridization and/or amplification reaction on mRNA contained in said urine sample, said first oligonucleotide or first primer pair being specific for a prostate cancer associated PCA3 mRNA molecule which is;
(i) a polynucleotide molecule comprising the sequence of SEQ ID NO;
9, 10 or 13;
(ii) a polynucleotide molecule that hybridizes under high stringency conditions to (i), wherein said high stringency conditions comprise a hybridization at 65°
C. in 6×
SSC or 5×
SSPE, 5×
Denhardt'"'"'s solution, 0.5% SDS and 100 μ
g/ml denatured carrier DNA and a washing at 65°
C. in 0.2×
SSC/0.1% SDS;
or(iii) a polynucleotide molecule fully complementary to (i) or (ii); and
(c) a second oligonucleotide or second primer pair for performing a second RNA hybridization and/or amplification reaction on mRNA contained in said urine sample, said second oligonucleotide or second primer pair being specific for a prostate-specific mRNA molecule,wherein at least one oligonucleotide in the composition is a labeled oligonucleotide, and the at least one labeled oligonucleotide includes at least one of (i) the first oligonucleotide, (ii) a primer of the first primer pair, (iii) the second oligonucleotide, or (iv) a primer of the second primer pair.
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Accused Products
Abstract
The present invention provides methods to detect prostate cancer by detecting the RNA encoded by PCA3. The disclosure provides a method for determining a predisposition, or presence of prostate cancer comprising: (a) contacting a sample with at least one oligonucleotide that hybridizes to a PCA3 polynucleotide; (b) detecting an amount of PCA3 and second prostate-specific polynucleotides; and (c) comparing the amount of PCA3 polynucleotide that hybridizes to the oligonucleotide to a predetermined cut off value, and determining the presence or absence of prostate cancer. Diagnostic kits are provided for detecting prostate cancer or the risk of developing same comprising: (a) at least one container means containing at least one oligonucleotide probe or primer that hybridizes to PCA3 (b) at least one oligonucleotide probe or primer that hybridizes with a second prostate specific nucleic acid; and (c) reagents for detecting PCA3 and the second prostate specific nucleic acid.
72 Citations
20 Claims
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1. A composition comprising:
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(a) a urine sample from a subject having or suspected of having prostate cancer, said urine sample comprising at least one prostate cell or nucleic acid extract thereof; (b) a first oligonucleotide or first primer pair for performing an RNA hybridization and/or amplification reaction on mRNA contained in said urine sample, said first oligonucleotide or first primer pair being specific for a prostate cancer associated PCA3 mRNA molecule which is; (i) a polynucleotide molecule comprising the sequence of SEQ ID NO;
9, 10 or 13;(ii) a polynucleotide molecule that hybridizes under high stringency conditions to (i), wherein said high stringency conditions comprise a hybridization at 65°
C. in 6×
SSC or 5×
SSPE, 5×
Denhardt'"'"'s solution, 0.5% SDS and 100 μ
g/ml denatured carrier DNA and a washing at 65°
C. in 0.2×
SSC/0.1% SDS;
or(iii) a polynucleotide molecule fully complementary to (i) or (ii); and (c) a second oligonucleotide or second primer pair for performing a second RNA hybridization and/or amplification reaction on mRNA contained in said urine sample, said second oligonucleotide or second primer pair being specific for a prostate-specific mRNA molecule, wherein at least one oligonucleotide in the composition is a labeled oligonucleotide, and the at least one labeled oligonucleotide includes at least one of (i) the first oligonucleotide, (ii) a primer of the first primer pair, (iii) the second oligonucleotide, or (iv) a primer of the second primer pair. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19. A composition comprising:
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(a) a urine sample from a subject having or suspected of having prostate cancer, said urine sample comprising at least one prostate cell or nucleic acid extract thereof; (b) a first primer pair including a first promoter-primer for performing an RNA amplification reaction on mRNA contained in said urine sample, the first promoter-primer being specific for a prostate cancer associated PCA3 mRNA molecule which is; (i) a polynucleotide molecule comprising the sequence of SEQ ID NO;
9, 10 or 13;(ii) a polynucleotide molecule that hybridizes under high stringency conditions to (i), wherein said high stringency conditions comprise a hybridization at 65°
C. in 6×
SSC or 5×
SSPE, 5×
Denhardt'"'"'s solution, 0.5% SDS and 100 μ
g/ml denatured carrier DNA and a washing at 65°
C. in 0.2 SSC/0.1% SDS;
or(iii) a polynucleotide molecule fully complementary to (i) or (ii); and (c) a second primer pair including a second promoter-primer for performing a second RNA amplification reaction on mRNA contained in said urine sample, said second oligonucleotide or second primer pair being specific for a prostate-specific mRNA molecule. - View Dependent Claims (20)
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Specification