Methods for developing urine biomarkers and for detecting bladder cancer

A method of identifying the presence or absence of bladder cancer in a subject comprising:

• determining the expression level of at least one gene selected from DHRS2, UPK2, UPK1A, KRT20, UPK1B, ACER2, SNX31, VGLL1, FER1L4, UPK3A, OSR1, GATA3, SCUBE2, GAPDH, ADAM15 or HBB in a biological sample from the subject, wherein the biological sample comprises at least one microvesicular nucleic acid extracted from at least one microvesicle;
• determining at least one reference value in the biological sample;
• normalizing the expression level of the at least one gene to the at least one reference value to obtain a normalized expression level of the at least one gene;
• comparing the normalized expression level of the at least one gene to a predetermined cutoff value;
• and
• identifying the presence of bladder cancer in the subject when the normalized expression level of the at least one gene is less than the predetermined cutoff value or identifying the absence of bladder cancer in the subject when the normalized expression level of the at least one gene is greater than or equal to the predetermined cutoff value.

A method of identifying the presence or absence of bladder cancer in a subject comprising:

• determining the expression level of a first and at least a second gene selected from DHRS2, UPK2, UPK1A, KRT20, UPK1B, ACER2, SNX31, VGLL1, FER1L4, UPK3A, OSR1, GATA3, SCUBE2, GAPDH, ADAM15 or HBB in a biological sample from the subject, wherein the biological sample comprises at least one microvesicular nucleic acid extracted from at least one microvesicle;
• determining Δexpression, wherein Δexpression is the difference between the expression level of the first gene and the expression level of the at least second gene;
• comparing Δexpression to a predetermined cutoff value;
• identifying the presence of bladder cancer in the subject when Δexpression is greater than the predetermined cutoff value or identifying the absence of bladder cancer in the subject when Δexpression is less than or equal to the predetermined cutoff.

A method of identifying the presence or absence of bladder cancer in a subject comprising:

• determining the expression level of a first and at least a second gene selected from DHRS2, UPK2, UPK1A, KRT20, UPK1B, ACER2, SNX31, VGLL1, FER1L4, UPK3A, OSR1, GATA3, SCUBE2, GAPDH, ADAM15 or HBB in a biological sample from the subject, wherein the biological sample comprises at least one microvesicular nucleic acid extracted from at least one microvesicle;
• determining Δexpression, wherein Δexpression is the difference between the expression level of the first gene and the expression level of the at least second gene;
• comparing Δexpression to a predetermined cutoff value;
• and
• identifying the presence of bladder cancer in the subject when Δexpression is less than the predetermined cutoff value or identifying the absence of bladder cancer in the subject when Δexpression is greater than or equal to the predetermined cutoff.

A method of identifying the presence or absence of bladder cancer in a subject comprising:

• determining the expression level of a first, a second gene and at least a third gene selected from DHRS2, UPK2, UPK1A, KRT20, UPK1B, ACER2, SNX31, VGLL1, FER1L4, UPK3A, OSR1, GATA3, SCUBE2, GAPDH, ADAM15 or HBB in a biological sample from the subject, wherein the biological sample comprises at least one microvesicular nucleic acid extracted from at least one microvesicle;
• determining at least one reference value in the biological sample;
• normalizing the expression level of the at least third gene to the at least one reference value to obtain a normalized expression level of the at least third gene;
• determining Δexpression, wherein Δexpression is the difference between the expression level of the first gene and the expression level of the at least second gene;
• comparing Δexpression to a first predetermined cutoff value;
• comparing the normalized expression level of the at least third gene to a second predetermined cutoff value;
• and
• identifying the presence of bladder cancer in the subject when Δexpression is greater than the first predetermined cutoff value and the normalized expression level of the at least third gene is greater than the second predetermined cutoff value or identifying the absence of bladder cancer in the subject when Δexpression is less than or equal to the first predetermined cutoff value and the normalized expression level of the at least third gene is less than or equal to the second predetermined cutoff value.

A method of identifying the presence or absence of bladder cancer in a subject comprising:

• determining the expression level of a first, a second gene and at least a third gene selected from DHRS2, UPK2, UPK1A, KRT20, UPK1B, ACER2, SNX31, VGLL1, FER1L4, UPK3A, OSR1, GATA3, SCUBE2, GAPDH, ADAM15 or HBB in a biological sample from the subject, wherein the biological sample comprises at least one microvesicular nucleic acid extracted from at least one microvesicle;
• determining at least one reference value in the biological sample;
• normalizing the expression level of the at least third gene to the at least one reference value to obtain a normalized expression level of the at least third gene;
• determining Δexpression, wherein Δexpession is the difference between the expression level of the first gene and the expression level of the at least second gene;
• comparing Δexpression to a first predetermined cutoff value;
• comparing the normalized expression level of the at least third gene to a second predetermined cutoff value;
• and
• identifying the presence of bladder cancer in the subject when Δexpression is greater than the first predetermined cutoff value and the normalized expression level of the at least third gene is less than the second predetermined cutoff value or identifying the absence of bladder cancer in the subject when Δexpression is less than or equal to the first predetermined cutoff value and the normalized expression level of the at least third gene is greater than or equal to the second predetermined cutoff value.

A method of identifying the presence or absence of bladder cancer in a subject comprising:

• determining the expression level of a first, a second gene and at least a third gene selected from DHRS2, UPK2, UPK1A, KRT20, UPK1B, ACER2, SNX31, VGLL1, FER1L4, UPK3A, OSR1, GATA3, SCUBE2, GAPDH, ADAM15 or HBB in a biological sample from the subject, wherein the biological sample comprises at least one microvesicular nucleic acid extracted from at least one microvesicle;
• determining at least one reference value in the biological sample;
• normalizing the expression level of the at least third gene to the at least one reference value to obtain a normalized expression level of the at least third gene;
• determining Δexpression wherein Δexpression is the difference between the expression level of the first gene and the expression level of the at least second gene;
• comparing Δexpression to a first predetermined cutoff value;
• comparing the normalized expression level of the at least third gene to a second predetermined cutoff value;
• and
• identifying the presence of bladder cancer in the subject when Δexpression is less than the first predetermined cutoff value and the normalized expression level of the at least third gene is greater than the second predetermined cutoff value or identifying the absence of bladder cancer in the subject when Δexpression is greater than or equal to the first predetermined cutoff value and the normalized expression level of the at least third gene is less than or equal to the second predetermined cutoff value.

A method of identifying the presence or absence of bladder cancer in a subject comprising:

• determining the expression level of a first, a second gene and at least a third gene selected from DHRS2, UPK2, UPK1A, KRT20, UPK1B, ACER2, SNX31, VGLL1, FER1L4, UPK3A, OSR1, GATA3, SCUBE2, GAPDH, ADAM15 or HBB in a biological sample from the subject, wherein the biological sample comprises at least one microvesicular nucleic acid extracted from at least one microvesicle;
• determining at least one reference value in the biological sample;
• normalizing the expression level of the at least third gene to the at least one reference value to obtain a normalized expression level of the at least third gene;
• determining Δexpression, wherein Δexpression is the difference between the expression level of the first gene and the expression level of the at least second gene;
• comparing Δexpression to a first predetermined cutoff value;
• comparing the normalized expression level of the at least third gene to a second predetermined cutoff value;
• and
• identifying the presence of bladder cancer in the subject when Δexpression is less than the first predetermined cutoff value and the normalized expression level of the at least third gene is less than the second predetermined cutoff value or identifying the absence of bladder cancer in the subject when Δexpression is greater than or equal to the first predetermined cutoff value and the normalized expression level of the at least third gene is greater than or equal to the second predetermined cutoff value.