Diagnosis and risk stratification of bladder cancer

A method of defining the likelihood of a subject having bladder cancer, comprising the steps of:

• (A) assessing the subject's likelihood of having bladder cancer by:
• i. identifying at least one sub-population group appropriate to the subject;
• ii. measuring the protein concentration level of one or more biomarkers selected according to the sub-population group in a sample obtained from the subject, wherein said measuring comprises contacting the sample with a substrate having at least one antibody against each of the one or more biomarkers on the substrate;
• iii. inputting one or more values into an algorithm to produce an output value, wherein the one or more inputted values are representative of the measured protein concentration level of the one or more biomarkers, and wherein the algorithm carries out ROC statistical analysis and/or logistical regression;
• and
• iv. correlating the output value with the likelihood of the subject having bladder cancer,
• wherein the sub-population group is selected according to smoking habits, gender, presence/absence of stone disease, history of benign prostate enlargement (BPE) or prescription of anti-hypertensive, anti-platelet and/or anti-ulcer medication, and
• (B) determining the subject's stratified risk level of serious disease by:
• v. measuring the protein concentration level of one or more biomarkers specific for one or more risk classifiers defined using Random Forest Classifiers (RFC), logistic regression or another appropriate systems biology or statistical approach in a sample obtained from the subject, wherein said measuring comprises contacting the sample with a substrate having at least one antibody against each of the one or more biomarkers on the substrate,
• vi. inputting one or more values into an algorithm or algorithms to produce an output value, wherein the one or more inputted values are representative of the measured protein concentration level of the one or more biomarkers, and wherein the one or more algorithms carry out ROC statistical analysis and/or logistical regression;
• and
• vii. correlating the output value with a stratified risk level of underlying serious disease,
• wherein the likelihood of having bladder cancer is combined with the stratified risk level of having serious disease, wherein the risk of having bladder cancer and/or serious disease is categorized as:
• high-risk bladder cancer requiring immediate cystoscopy;
• low-risk bladder cancer requiring urgent cystoscopy;
• high-risk control requiring close evaluation and further investigation;
• or low-risk control requiring primary care monitoring, and wherein:
• if the smoker sub-population is selected, the protein concentration level of the biomarkers CRP, EGF, MMP9, IL-1α, IL-4, TM and IL-2 is measured in step (ii);
• if the non-smoker sub-population is selected, the protein concentration level of the biomarkers TNFα, sTNFR1, IL-6, IL-1α, MMP9/NGAL complex and CEA is measured in step (ii) and the creatinine protein concentration level in the sample is also measured;
• if the gender sub-population is selected and the subject is male, the protein concentration level of the biomarkers CRP, EGF, CK18, IL-1β, IL-8 and IL-2 is measured in step (ii) and the creatinine protein concentration level in the sample is also measured;
• if the gender sub-population is selected and the subject is female, the protein concentration level of the biomarkers CRP, EGF, IL-6, dDimer, MMP9/NGAL complex and CEA is measured in step (ii) and the osmolarity of the sample is also measured;
• if the stone disease sub-population is selected and the subject is positive for stone disease, the protein concentration level of the biomarkers CRP, sTNFR1, CK18, IL-1α, IL-8 and VEGF is measured in step (ii) and the creatinine protein concentration level in the sample is also measured;
• if the stone disease sub-population is selected and the subject is negative for stone disease, the protein concentration level of the biomarkers CRP, EGF, IL-6, IL-1α, MMP9/NGAL complex and CEA is measured in step (ii) and the creatinine protein concentration level in the sample is also measured;
• if the BPE sub-population is selected and the subject is positive for BPE, the protein concentration level of the biomarkers CRP, EGF, IL-6, IL-1α, MMP9/NGAL complex, TM and CEA is measured in step (ii);
• if the BPE sub-population is selected and the subject is negative for PBE, the protein concentration level of the biomarkers CRP, EGF, CK18, NGAL, MMP9/NGAL complex and BTA is measured in step (ii) and the creatinine protein concentration level in the sample is also measured;
• if the anti-hypertensive medication sub-population is selected and the subject is positive for anti-hypertensive medication, the protein concentration level of the biomarkers TNFα, EGF, IL-6, MMP9/NGAL complex and CEA is measured in step (ii) and the creatinine protein concentration level and total protein level in the sample is also measured;
• if the anti-hypertensive medication sub-population is selected and the subject is negative for anti-hypertensive medication, the protein concentration level of the biomarkers TNFα, sTNFR1, IL-6, NGAL, IL-8, TM and CEA is measured in step (ii);
• if the anti-platelet medication sub-population is selected and the subject is positive for anti-platelet medication, the protein concentration level of the biomarkers TNFα, EGF, IL-6, IL-8 and CEA is measured in step (ii) and the total protein level and osmolarity of the sample is also measured;
• if the anti-platelet medication sub-population is selected and the subject is negative for anti-platelet medication, the protein concentration level of the biomarkers CRP, EGF, MCP-1, MMP9/NGAL complex, TM and FPSA is measured in step (ii) and the total protein level of the sample is also measured;
• if the anti-ulcer medication sub-population is selected and the subject is positive for anti-ulcer medication, the protein concentration level of the biomarkers CRP, EGF, IL-6, IL-1α, IL-8, TM and CEA is measured in step (ii);
• and
• if the anti-ulcer medication sub-population is selected and the subject is negative for anti-ulcer medication, the protein concentration level of the biomarkers CRP, EGF, vWF, IL-1β, MMP9/NGAL complex, TM and HA is measured in step (ii).