Urinary WT1-positive cells as a non-invasive biomarker of crescent formation.

OBJECTIVE: The purpose of this study was to assess the relationship between urinary WT1-positive cells (podocytes and active parietal epithelial cells) and WT1-positive cells in renal biopsy to investigate whether urinary WT1-positive cells are useful for detection of crescent formation. METHODS: Fifty-two patients with kidney disease were investigated (15 cases with crescentic lesions and 37 cases with non-crescentic lesions) for immunoenzyme staining using anti-WT1 antibody for urine cytology and renal biopsy. Numbers of WT1-positive cells in urine and renal biopsy were counted. RESULTS: There was no correlation between urinary WT1-positive cells and WT1-positive cells in renal biopsy. However, the number of urinary WT1-positive cells in patients with crescentic lesions was significantly higher than in patients with non-crescentic lesions. In addition, the best cut-off value to detect patients with crescentic lesions using urinary was 5 cells/10-mL (area under the concentration-time curve=0.735). CONCLUSIONS: The results of our study suggest urinary WT1-positive cells can be used to detect patients with crescent formation using 5 cells/10-mL cutoff value. WT1-positive glomerular podocytes and parietal epithelial cells may be shed into urine in active glomerular disease. This study, investigating the relationship between WT1-positive cells in urine and in the renal biopsy found no correlation; however, the results do suggest that, using a cutoff value of 5 cells/10 mL, WT1 positive urinary cells can be used to detect patients with crescent formation.

Expression of vimentin and high-molecular-weight cytokeratin (clone 34ßE12) in differentiating reactive renal tubular cells from low-grade urothelial carcinoma cells in voided urine.

OBJECTIVE: Reactive renal tubular cells show features of an atypical repair reaction. Differentiation between reactive renal tubular cells and low-grade urothelial carcinoma (LG-UC) cells can therefore be a diagnostic challenge based on morphology alone. In this study, we evaluated the diagnostic utility of vimentin and a high-molecular-weight cytokeratin antibody (clone 34ßE12) in differentiating reactive renal tubular cells from LG-UC. METHODS: We evaluated voided urine cytology and surgical specimens from 40 patients with renal disease, and 17 patients with LG-UC. All slides were stained with vimentin and 34ßE12. RESULTS: In the reactive renal tubular cells in voided urine cytology, vimentin showed strong cytoplasmic staining in 39/40 (97.5%) cases, but all were negative for 34ßE12. LG-UC cells showed positive staining for 34ßE12 in 3/17 (17.6%) cases, whereas none were positivity for vimentin. The reactive renal tubular cells of histological specimens in the renal disease group demonstrated positive for vimentin in all 40 cases and all were negative for 34ßE12. The LG-UC group showed abnormal staining for 34ßE12 in 4/17 (23.5%) cases, whereas none were positive for vimentin. CONCLUSIONS: Vimentin expression in urine cytology can help to distinguish reactive renal tubular cells from LG-UC. However, 34ßE12 does not appear to be a useful adjunct to distinguish these two groups in voided urine cytology.

Focus on urinary bladder cancer markers: a review.

Finding and development of new bladder cancer markers is still a very dynamic field. Because of the mass of all these markers it is impossible to report all of them. This paper reviews the role of bladder cancer markers in diagnosis and highlights the most important biomarkers studied and reported recently. A medline based literature search was performed to examine the field of bladder cancer markers. Major topics focus on selected bladder cancer markers from nearly all categories of the wide field of bladder cancer markers: Hematuria, FISH, FGFR3, SURVIVIN, u-PAR, TP53 mutation, HER-2/neu, TPA, NMP22, CK-19, CK-20, CYFRA 21-1. The use and clinical importance as diagnostic help are discussed. In this review a highlight to some of the most important markers was made. Further determination of recurrence and progression marker will contribute to establish better treatments for the individual patient. Molecular staging of urological tumors will allow selecting cases that will require systemic treatment. It is necessary and important to integrate under the same objectives basic and clinical research.

Evaluation of the diagnostic accuracy of UBC® Rapid in bladder cancer: a Swedish multicentre study.

OBJECTIVE: The aim of this study was to determine the diagnostic accuracy of UBC® Rapid - a urine-based marker for bladder cancer - in patients with bladder cancer and controls, and to compare the test results across risk groups. MATERIALS AND METHODS: This prospective phase II study was conducted at four Swedish hospitals. UBC Rapid was evaluated in four groups: A, newly diagnosed bladder cancer (n = 94); B, follow-up of non-muscle-invasive bladder cancer (n = 75); C, benign urinary tract diseases (n = 51); and D, healthy controls (n = 50). Tumours were divided into high risk (carcinoma in situ, TaG3, T1, T2 and T3) and low risk (low malignant potential, TaG1 and TaG2). Urine samples were quantitatively analysed by UBC Rapid. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were calculated based on optimal cut-off (receiver operator characteristics curve analysis). A linear regression compared the UBC Rapid results in the different risk groups. RESULTS: The optimal cut-off was 8.1 µg/l. The median UBC Rapid values were 9.3 µg/l [interquartile range (IQR) 30.9] and 4.3 µg/l (IQR 7.8) in patients with positive and negative cystoscopy, respectively (p < .001). The value for group A was 15.6 µg/l (IQR 37.9), group B 5.6 µg/l (IQR 8.6), group C 5.1 µg/l (IQR 9.0) and group D 3.3 µg/l (IQR 7.1). Sensitivity was 70.8%, specificity 61.4%, PPV 71.3% and NPV 60.8%. The high-risk group had significantly higher UBC Rapid values than the low-risk group: 20.5 µg/l (IQR 42.2), sensitivity 79.2% and specificity 61.4% versus 7.0 µg/l (IQR 9.9), sensitivity 60.0% and specificity 61.4% (p = .039). CONCLUSIONS: The UBC Rapid urine-based marker for bladder cancer gave higher values in patients with positive than in those with negative cystoscopy. The diagnostic accuracy was better in patients with high-risk than in those with low-risk tumours, and was better during primary detection than during surveillance.

Prognostic value of cytology, nuclear matrix protein 22 (NMP22) test, and urinary bladder cancer II (UBC II) test in early recurrent transitional cell carcinoma of the bladder.

This study addresses the diagnostic value of the Nuclear Matrix Protein 22 (NMP22) test and the Urinary Bladder Cancer (UBC II) test, in comparison to bladder wash cytology for the detection of early recurrence of bladder cancer. Patients with transitional cell carcinoma of the bladder (TCC, n = 60) and patients with benign urological diseases (n = 30) were included in this study. Voided urine samples were divided into 2 aliquots: aliquot 1 was assayed for NMP22 and aliquot 2 was tested for UBC II. Saline bladder washings were used for cytologic examination. Urine samples from TCC patients were collected before transurethral resection and on postoperative day 10. On day 10, 15 NMP22 results and 7 UBC II results exceeded the normal ranges; 4 of the cytology samples were positive for malignancy. Based on cystoscopic findings at 3 mo post-resection, 21 of the cases were classified as early recurrence; 11 of the early recurrences had been in the elevated NMP22 group, 4 in the elevated UBC II group, and 3 in the positive cytology group at 10 days post-resection. The NMP22 test gave the highest sensitivity for detecting early recurrent tumors (11 of 21, 52%). Such high sensitivity did not occur with the UBC II test (4 of 21, 19%) or cytology (3 of 21, 14%). These differences were significant (p = 0.024 and 0.009, respectively). Thus, the NMP22 test showed superiority over the other tests for detection of early recurrence of bladder cancer.

Pre and postoperative quantitative detection of fragments of cytokeratins 8 and 18 (UBC IRMA) as markers of early recurrence of superficial bladder tumor.

OBJECTIVES: The Urinary Bladder Cancer (UBC) test is a marker that detects urinary fragments of cytokeratin 8 and 18. The aim of this study is to evaluate the usefulness of the pre and post operative UBC test to detect early recurrences of a bladder tumor in the first year after the transurethral resection of a bladder tumor. MATERIALS AND METHODS: A multicentric perspective study on 36 patients with superficial bladder cancer (pTa-pT1) treated with transurethral resection (TUR) was performed. Each patient underwent 4 specific urine collections: 1) preoperatively, 2) 3 days after TUR, 3) 7 days after TUR, 4) 30 days after TUR. UBC was analysed on urine with the IRMA method and the cut off value of 12 mg/L was used. Cystoscopy was performed after 3, 6, 9, and 12 months after TUR, with the aim of identifying all cancer recurrences in the first year postoperatively. Statistical analyses to identify differences between patients with or without early recurrence were performed in accordance with Fisher's exact test and Chi-square analysis. RESULTS: Of the 36 patients included in the study 15 showed early recurrence and 21 were recurrence free 1 year after surgery. UBC levels measured in recurrence free patients 30 days after TUR showed normal values, values decreasing as compared with preoperative levels or both circumstances, even if a statistically significant difference was not found between the two groups. CONCLUSIONS: In this study we reported an insignificant correlation between the postoperative modifications of UBC levels and the risk of tumor recurrence during the first year of follow-up. A larger study group with longer follow-ups will probably allow better evaluation of the real power of UBC tests in clinical practice.

Evaluation of two new urinary tumor markers: bladder tumor fibronectin and cytokeratin 18 for the diagnosis of bladder cancer.

Our objectives were to evaluate the diagnostic value of two new urinary tumor markers, cytokeratin 18 (CK18) and bladder tumor fibronectin (BTF), for the detection and monitoring of bladder cancer. The study comprised 931 urine samples belonging to 402 subjects: 112 individuals under suspicion for a primary bladder tumor (group 1); 104 bladder cancer patients under scheduled follow-up (group 2); 109 bladder cancer patients receiving intravesical instillations (group 3); 45 patients with other urological diseases (group 4); and 32 healthy subjects (group 5). Voided urine samples were collected before cystoscopies, between them and before intravesical instillations. CK18 and BTF tests were measured by chemiluminescent immunoassays. Optimal receiver operating characteristic cutoffs of 7.4 microg/L for CK18 and 52.8 microg/liter for BTF rendered overall sensitivities of 66.2% for CK18 and 80.0% for BTF at specificities of 88.4 and 74.7%, respectively. Urinary cytology provided a sensitivity of 29.2% at a specificity of 99.1%. Sensitivities were 80.8, 74.2, and 82.3% for BTF and 71.1, 77.4, and 64.7% for CK18 for groups 1 to 3, respectively. False positive rates were higher for BTF in all groups of patients. Elevated urinary tumor markers during the monitoring of patients with bladder cancer could detect recurrence sooner than scheduled cystoscopies. Persistence of negative markers was greatly indicative of free of disease status in follow-up. CK18 and BTF in urine may eventually prove to be of benefit for specific patients with bladder carcinoma given its higher sensitivity compared with cytology. In selected patients, namely those with persistent negative urinary CK18 and BTF, the number of cystoscopies could be reduced.

Evaluation of the UBC test in the urine of healthy individuals, patients with benign disorders and urinary bladder cancer.

Using the UBC test, the specificity, sensitivity and prognostic information were evaluated in patients with recently diagnosed transitional cell carcinoma (TCC) and in a control group consisting of apparently healthy individuals and individuals with benign disorders. Frozen urine samples from the 485 individuals in the control group and 100 newly diagnosed TCC patients were analyzed with the UBC test, specific for epitopes on cytokeratin fragments released from the urothelial cells. All the samples were analyzed and corrected for creatinine. No significant concentration difference was found between males and females (p=0.65) and there was no age dependent relation. The median concentration for the entire control group was estimated at 3.7 microg/g and the 95th percentile was calculated at 53.0 microg/g. The apparently healthy individuals in the control group had a median value of 3.4 microg/g with a 95th percentile of 24.3 microg/g. An increased frequency of elevated UBC concentrations was found in some benign disorders e.g., anemia, thyroid disorders, diabetes mellitus, hyperlipemia, urosepsis and cystitis. Patients with superficial tumors exhibited a 66% sensitivity (at 95% specificity), and the UBC concentrations did not differ statistically (p=0.16) from those patients with muscle invasive lesions with a 52% sensitivity. When the UBC concentrations were related to histopathological grade, a significant concentration difference (p<0.004) was found between low grade tumors (sensitivity 41%) and high grade tumors (sensitivity 72%). Survival analysis showed that patient with muscle invasive tumors, high-grade tumors and high UBC concentrations have a significantly reduced survival (five-year survival was estimated to 30%, 35% and 30% respectively) compared to patients with superficial tumors, low-grade tumors or low UBC concentrations (five-year survival, 60%, 85% and 75% respectively). The UBC test showed good accuracy and repeatability. Clinically the test could assist in tumor grading and the detection of recurrent disease, which in turn could assist in treatment selection for the individual patient and possibly improve prognosis.

The clinical value of CYFRA21-1 in bladder cancer patients: Egyptian experience.

There are a wide variety of tumor markers now available that proved to be of value in the management of cancer patients. Of these markers, tissue polypeptide antigen (TPA) and tissue polypeptide specific antigen (TPS) are well known in the field of bladder cancer. TPA was found to be a mixture of cytokeratins 8, 18 and 19 and recent investigations proved that TPS is keratin 18. The aim of the present study was to assess the clinical value of urinary cytokeratin 19 (CYFRA21-1) in the differential diagnosis between bladder cancer and benign urinary tract diseases represented by bilharziasis. Two hundreds and seventy individuals were included in the present study: 186 with bladder cancer representing the different stages and grades, 44 with urinary tract bilharziasis and 40 normal healthy controls. CYFRA21-1 was evaluated in 24-hour urine samples by ELISA using the automatic set supplied by Boehringer Manheim, Manheim, Germany (ES 300). Results of this study revealed significant elevation of CYFRA21-1 in bladder cancer followed by bilharziasis. 82.3% (153/186) of bladder cancer patients and 11.4% (5/44) of bilharzial patients exhibited CYFRA21-1 levels above the upper limit of the control group (3 micrograms/24-hr). CYFRA21-1 was more sensitive in advanced than early stages of bladder cancer and in patients with positive than those with negative lymph nodes, but association of tumor with bilharziasis did not markedly affect its level.

[Cytokeratins (UBC and CYFRA 21-1) and nuclear matrix proteins (NMP22) as urine tumor markers in the diagnosis of bladder cancer]. / Citoqueratinas (UBC y CYFRA 21-1) y proteínas de la matriz nuclear (NMP22) como marcadores tumorales en la orina en el diagnóstico del cáncer vesical.

BACKGROUND: The development of urinary tumor markers such as UBC, CYFRA 21-1 and NMP22 appeared to be non invasive alternative methods for the detection of bladder cancer. We compared the individual and combined sensitivity of the urinary tumor markers in the detection of bladder cancer, contrasting them with the conventional diagnostic procedures. PATIENTS AND METHODS: 237 voided urines from subjects under risk for bladder cancer were collected immediately before the endoscopic examinations: 44 patients under suspicion of a primary bladder tumor and 193 patients under follow-up of a previous bladder cancer were included. UBC and NMP22 were measured by enzyme-immunoabsorbent-assays and CYFRA 21-1 by an electro-chemiluminescense-immunoassay. RESULTS: Taking the cutoffs of 9.7 micrograms/l for UBC, 5.4 ng/ml for CYFRA 21-1 and 10.0 U/ml for NMP22 sensitivities were 70%, 69% and 67% for UBC, CYFRA 21-1 and NMP22 at specificities of 95%, 94% y 80%, respectively. All tumor markers showed higher sensitivities than urinary cytology (7%), microhematuria (62%) and gross hematuria (10%) at specificities of 99%, 78% and 99%, respectively. The combinations of NMP22 plus CYFRA 21-1 reached the highest sensitivity (79%), slightly lower than simultaneously measuring the three tumor markers (80%). CONCLUSIONS: The sensitivities of the urinary markers UBC, CYFRA 21-1 and NMP22 appeared to be high enough so as to substitute urinary cytology. The diagnostic similarity between cytokeratins individually and in each type of patients might not recommend their simultaneous determination. The combined measurement of NMP22 and one cytokeratin marker (CYFRA 21-1 or UBC) appeared to be the most recommended.

[Differential diagnosis of kidney transplant rejection and ciclosporin nephrotoxicity by urine cytology].

To make the differentiation of kidney transplant acute rejection and ciclosporin (CS) nephrotoxicity urine cytology by classical Papanicolaou with immunocytochemical stain has been performed. Increased numbers of renal tubular cells with lymphocytes and monocytes were found in both rejections and CS toxicities. CS toxicities were associated with increased numbers of proximal tubular cells. In immunocytochemical stain, increased numbers of CD25 and CD8 positive cells as well as increased ratio of HLA-DR/cytokeratin positive cells were typically found in rejections. It is concluded that the proposed analysis of urine cytology is a non-invasive and reliable method for daily graft monitoring of acute rejection and CS toxicity.

[The clinical usefulness of urinary determinations of cytokeratin 19 fragment (CYFRA 21-1) in urothelial tumor].

The urinary CYFRA 21-1 value corrected for urinary creatinine (ng/ml/creatinine), was studied in the urine of patients with urothelial tumors. To examine its clinical significance we studied urinary CYFRA 21-1 excretion (ng/ml/creatinine), in a total of 22 urine samples from patients with bladder cancer, 7 from patients with renal pelvic and ureteral tumor, 6 from patients with urinary infection 6 from patients with urinary diversion called ileal conduit, and 8 from healthy adult men. The excretion of CYFRA 21-1 in urine was determined by two specific monoclonal antibodies (Ks 19.1 and BW 19.21). The mean value of urinary CYFRA 21-1 in healthy adult men was 1.96 +/- 1.33 (mean +/- SD) ng/ml/creatinine. Urinary CYFRA 21-1 showed a higher value in the urine of urinary infection and urinary diversion. As to bladder cancer, urinary CYFRA 21-1 showed a higher value in a larger volume of tumor than in a smaller volume of tumor in transitional cell carcinoma regardless of the grade and stage. These findings suggest that urinary CYFRA 21-1 may be a non-specific marker in urothelial tumors.

Biomarker analysis of Morquio syndrome: identification of disease state and drug responsive markers.

BACKGROUND: This study was conducted to identify potential biomarkers that could be used to evaluate disease progression and monitor responses to enzyme replacement therapy (ERT) in patients with mucopolysaccharidosis (MPS) IVA. METHODS: Levels of 88 candidate biomarkers were compared in plasma samples from 50 healthy controls and 78 MPSIVA patients not receiving ERT to test for significant correlations to the presence of MPSIVA. MPSIVA samples were also tested for correlations between candidate biomarkers and age, endurance, or urinary keratin sulfate (KS) levels. Then, levels of the same 88 analytes were followed over 36 weeks in 20 MPSIVA patients receiving ERT to test for significant correlations related to ERT, age, or endurance. RESULTS: Nineteen candidate biomarkers were significantly different between MPSIVA and unaffected individuals. Of these, five also changed significantly in response to ERT: alpha-1-antitrypsin, eotaxin, lipoprotein(a), matrix metalloprotein (MMP)-2, and serum amyloid P. Three of these were significantly lower in MPSIVA individuals versus unaffected controls and were increased during ERT: alpha-1-antitrypsin, lipoprotein(a), and serum amyloid P. CONCLUSIONS: Candidate biomarkers alpha-1-antitrypsin, lipoprotein(a), and serum amyloid P may be suitable markers, in addition to urinary KS, to follow the response to ERT in MPSIVA patients.

Evaluation of urine tumor-associated trypsin inhibitor, CYFRA 21-1, and urinary bladder cancer antigen for detection of high-grade bladder carcinoma.

OBJECTIVES: To assess the value of urine tumor-associated trypsin inhibitor (TATI), CYFRA 21-1, which measures cytokeratin 19 fragment, and urinary bladder carcinoma antigen (UBC) for the detection of high-grade bladder carcinoma. METHODS: A total of 160 individuals were enrolled in the present study. Of these, 80 were patients with proven primary high-grade urothelial bladder cancer (group 1), 40 were healthy volunteers (group 2), and 40 had history of benign urologic disease (group 3). All were evaluated with respect to urinary TATI, CYFRA 21-1, and UBC levels. All these markers were evaluated using commercial kits. Cytology was also performed. RESULTS: The TATI measurements were significant greater in group 1 compared with groups 2 and 3. The cutoff point used for TATI, CYFRA 21-1, and UBC was 22, 2.8, and 12 microg/L, respectively. The overall sensitivity was 85.7% for TATI, 61.9% for CYFRA 21-1, 50% for UBC, and 42.8% for cytology. TATI was significantly more sensitive in Stage Ta (80%) than was CYFRA 21-1 (32%), UBC (12%), and cytology (20%). TATI was also more sensitive compared with other tumor markers for Stage T1 but not for Stage T2 or T3. CONCLUSIONS: The results of our study have shown that TATI is a promising urinary tumor marker for high-grade urothelial bladder cancer. It is more sensitive than CYFRA 21-1, UBC, and cytology for Stage Ta and T1 bladder cancer.

Urinary CYFRA 21.1 is not a useful marker for the detection of recurrences in the follow-up of superficial bladder cancer.

OBJECTIVES: The objective of this prospective study is to establish an appropriate cutoff value of urinary CYFRA 21.1 assay and to assess its utility combined with voided cytology and/or haemoglobin dipstick in the follow-up of patients with superficial bladder cancer. METHODS: From December 2000 to November 2003, 446 patients in follow-up for superficial bladder cancer (Ta-T1) after transurethral resection of the bladder (TURB) were included in a prospective study. Voided urine specimens were collected 7-14 d before cystoscopy and/or TURB for CYFRA 21.1 (one sample), haemoglobin dipstick (one sample), and cytology (three samples). All samples were processed for CYFRA 21.1 and haemoglobin dipstick according to manufacturer instructions. A control group (n=185) was obtained from patients in follow-up after transurethral resection of superficial disease (without recurrences within the following 6 mo). There were 125 recurrent transitional tumours detected by cystoscopy (34 TaG1; 53 TaG2/T1G1-2; 23 Ta-1G3/Tis, and 15 T2-4). Receiver operator characteristic (ROC) curves were constructed and cutoff values were chosen. Sensitivity, specificity, PPV (positive predictive value), NPV (negative predictive value), and their 95% confidence intervals were calculated. RESULTS: ROC curve analysis based on the previously reported cutoff value of 4ng/ml for CYFRA 21.1 demonstrated a sensitivity and specificity of 43% and 68%, respectively. At a cutoff value of 1.5ng/ml, sensitivity was 73.8% with a low specificity (41%). Further lowering of the cutoff point below 1.5ng/ml did not demonstrate a significant increase in sensitivity. Therefore, this value was chosen as the most sensitive CYFRA 21.1 cutoff point during the rest of the study. Specificity increased when all the patients treated with pelvic radiotherapy or with UTI, urethral catheterisation, and intravesical instillations within 3 previous months were not included in our analysis. CYFRA 21.1 plus cytology and the combination of CYFRA 21.1, cytology, and haemoglobin dipstick demonstrated the highest overall sensitivities, and detected 91.3% of Ta-1G3 tumours and 93.3% of T2-4 tumours. However, there were one muscle-invasive tumour, two T1G3/Tis, three T1G2, and nine T1G1 neoplasms with negative combination of cytology and CYFRA 21.1 (1,5ng/ml). All these tumours were smaller than 2cm in size; most were single tumours. Nevertheless, there were 16 tumours larger than 0.5cm (0.5-2cm), and multiple neoplasms were endoscopically detected in 14 patients. Similar results were obtained through the combination of CYFRA 21.1 (cutoff: 1.5ng/ml), cytology, and haemoglobin dipstick. CONCLUSIONS: In our experience the low sensitivity of urinary CYFRA 21.1, even using lower cutoff values and/or a combination with cytology and/or haemoglobin dipstick, makes its application not very useful as a surveillance tool for superficial bladder carcinoma.

Combined assay of CYFRA21-1, telomerase and vascular endothelial growth factor in the detection of bladder transitional cell carcinoma.

BACKGROUND: CYFRA21-1, telomerase and vascular endothelial growth factor (VEGF) are regarded as useful tumor markers in the detection of bladder transitional cell carcinoma. However, the sensitivity of each single marker seemed to be unsatisfactory. In the current study, we assessed the sensitivity of the combined assay of these three markers in the detection of human bladder transitional cell carcinoma. METHODS: Voided urine samples of 100 patients with bladder transitional cell carcinoma were obtained and each sample was aliquoted for the various assay. Urinary CYFRA21-1 and VEGF were detected by enzyme-linked immunosorbent assay and telomerase detected by the telomeric repeat amplification protocol. The sensitivity of combined assay was compared to that of each single marker and urinary cytology, respectively. RESULTS: In the 100 patients, the sensitivity was 74% for urinary CYFRA21-1, 71% for telomerase, 69% for VEGF, and 38% for cytology. CYFRA21-1, telomerase and VEGF proved significantly more sensitive than cytology, respectively (P = 0.000). The sensitivity of the combined assay in the current study was 94% and it was significantly higher than that of cytology, urinary telomerase, CYFRA21-1 or VEGF (P = 0.000). In 50 patients with hematuria but without bladder cancer, the overall specificity of the assays was 78% for CYFRA21-1, 84% for telomerase, 88% for VEGF, and 92% for cytology. CONCLUSIONS: Combined assay of the markers which show different characteristics of the tumor behaviors seemed to be of interest in the detection of bladder transitional cell carcinoma.

Rapid diagnosis and follow up of bladder cancer patients using urinary high molecular weight cytokeratins.

We have developed an office-based dot-EIA for the detection of a urinary high molecular weight cytokeratin (CK). Immunohistochemical staining and western blot based on CK1K10 monoclonal antibody were used to identify the CK. Urine of 192 patients with different types, grades, and stages of bladder tumor and 72 controls were evaluated using dot-EIA. An intense and diffuse cytoplasmic reaction was shown in bladder squamous cell carcinoma. The target epitope was identified in urine at 65, 56, and 40-kDa. The CK purified from urine showed single polypeptide at 65-kDa using SDS-PAGE and single peak at 7.4 min using capillary zone electrophoresis. The dot-EIA detected the CK with high sensitivity (97%) and specificity (94%). The CK was not detected in urine of bladder cancer patients showing response to radiotherapy. The sensitive and specific office-based detection of urinary cytokeratin would be helpful in rapid diagnosis and follow up of bladder carcinoma.