Urine markers for detection of bladder cancer
Early detection of tumors is a major determinant of survival of patients suffering from tumors, including bladder tumors. Members of the BTM or UBTM family can be highly and consistently accumulated in bladder tumor tissue and other tumor tissue, and/or can be accumulated in urine of patients, and thus are markers for bladder and other types of cancer. In certain embodiments, BTMs or UBTMs can accumulate in the urine, and detection of UBTM family members can be an effective diagnostic approach. In some embodiments, quantitative PCR methods have advantages over microarray methods. In other embodiments, detection and quantification of a plurality of BTMs or UBTMs can increase the sensitivity and specificity of detection of bladder cancer, and therefore provides methods for determining the stage and type of bladder cancer. Kits provide easy, convenient ways for carrying out the methods of this invention.
1. A polymerase chain reaction (PCR) based kit for detecting bladder cancer, consisting of a forward primer, a reverse primer, and a labeled probe for each of the markers, sperm associated antigen 5 (SPAG5), topoisomerase (DNA) II alpha 170 kDa (TOP2A), cell division cycle 2, G1 to S and G2 to M (CDC2), endoglin (Osler-Rendu-Weber syndrome 1) (ENG), insulin-like growth factor binding protein 5 (IGFBP5), nephroblastoma overexpressed gene (NOV), neuropilin 1 (NRP1), sema domain, immunoglobulin domain (Ig), short basic domain, secreted, (semaphorin) 3F (SEMA3F), EGF-like-domain, multiple 6 (EGFL6), matrix Gla protein (MGP), semaphorin (SEM2), chromogranin A (parathyroid secretory protein 1) (CHGA), Thy-1 cell surface antigen (THY1), ubiquitin-conjugating enzyme E2C (USE2G), homeo box A13 (HOXA13), midkine (neurite growth-promoting factor 2) (MDK), baculoviral IAP repeat-containing 5 (survivin) (BIRC5), and SMC4 structural maintenance of chromosomes 4-like 1 (yeast) (SMC4L1); and
solutions for mixing reagents; and
reaction chambers for carrying out PCR amplification of each marker.
2. The polymerase chain reaction (PCR) based kit of claim 1 , wherein:
(a) at least two of the following combinations of oligonucleotides (i-xiii) are selected from the group consisting of:
(i) SEQ ID NO.1, SEQ ID NO.14, and SEQ ID NO.27;
(ii) SEQ ID NO.2, SEQ ID NO.15, and SEQ ID NO.28;
(iii) SEQ ID NO.3, SEQ ID NO.16, and SEQ ID NO.29;
(iv) SEQ ID NO.4, SEQ ID NO.17, and SEQ ID NO.30;
(v) SEQ ID NO.5, SEQ ID NO.18, and SEQ ID NO.31;
(vi) SEQ ID NO.6, SEQ ID NO.19, and SEQ ID NO.32;
(vii) SEQ ID NO.7, SEQ ID NO.20, and SEQ ID NO.33;
(viii) SEQ ID NO.8, SEQ ID NO.21, and SEQ ID NO.34;
(ix) SEQ ID NO.9, SEQ ID NO.22, and SEQ ID NO.35;
(x) SEQ ID NO.10, SEQ ID NO.23, and SEQ ID NO.36;
(xi) SEQ ID NO.11, SEQ ID NO.24, and SEQ ID NO.37;
(xii) SEQ ID NO.12, SEQ ID NO.25, and SEQ ID NO.38;
(xiii) SEQ ID NO.13, SEQ ID NO.26, and SEQ ID NO.39; and
(b) reaction reagents and reaction chambers for carrying out PCR amplification each of said combinations of oligonucleotides.
3. A polymerase chain reaction (PCR) based kit for detecting bladder cancer, consisting of combinations of oligonucleotides being a forward primer, a reverse primer, and a labeled probe for each of the markers, said markers consisting of:
a) cell division cycle 2, G1 to S and G2 to M (CDC2), insulin-like growth factor binding protein 5 (IGFBP5), homeo box A13 (HOXA13), and midkine (neurite growth-promoting factor 2) (MDK); and
b) reaction reagents and reaction chambers for carrying out PCR amplification each of said combinations of oligonucleotides.
4. The kit of claim 3 , said oligonucleotides selected from the group consisting of:
(i) SEQ ID NO.3 SEQ ID NO.16, and SEQ ID NO.29; and
(ii) SEQ ID NO.5, SEQ ID NO.18, and SEQ ID NO.31.
5. A polymerase chain reaction (PCR) based kit for detecting bladder cancer, consisting of combinations of oligonucleotides being a forward primer, a reverse primer, and a labeled probe for each of the markers, said markers consisting of
a) homeo box A13 (HOXA13), and midkine (neurite growth-promoting factor 2) (MDK); and
b) reaction reagents and reaction chambers for carrying out PCR amplification each of said combinations of oligonucleotides.