Improved detection of urothelial carcinomas with fluorescence immunocytochemistry (uCyt+ assay) and urinary cytology: results of a French Prospective Multicenter Study

The aim of the study was to assess the sensitivity and specificity of fluorescence immunocytochemistry (uCyt+ assay) as combined with urinary cytology for detection of primary and recurrent urothelial carcinomas. We analyzed 694 urinary samples from 236 new symptomatic patients and 458 patients followed after transurethral resection (TUR) for bladder tumor. Lesions suspicious for cancer at cystoscopy were sampled by biopsies or TUR. Sensitivity and specificity of tests were calculated using cystoscopy and histopathology, whether or not combined as gold standards. In new symptomatic patients, sensitivity of uCyt+ was 40%, 88.2%, and 76.7%, whereas that of urinary cytology was 30%, 70.6%, and 83.3%, respectively, in G1, G2, and G3 tumors. In follow-up cases, sensitivity of uCyt+ was 61.9%, 66.7%, and 76.9%, whereas that of urinary cytology was 38.1%, 58.3%, and 64.1%, respectively, in G1, G2, and G3 tumors. The combination of uCyt+ and urinary cytology significantly increased mean sensitivity in newly diagnosed cases (86.4% versus 71.2% with urinary cytology only, p < 0.05), as well as in patients followed after TUR (79.3% versus 55.2%, p < 0.001). Specificity of uCyt+ and urinary cytology was identical in new patients (83.3%) and was 81.9% and 86.2%, respectively, in patients followed after TUR. In patients with negative cystoscopy, positive uCyt+ tests had a strong predictive value for tumor recurrence at 1 year (47.0% versus 11.9% in patients with negative assay, p < 0.01). We conclude that combining uCyt+ with urinary cytology improves the detection of urothelial carcinomas as well in patients with symptoms suggesting bladder cancer as in those followed after treatment.     

Utility of the BTA stat test kit for bladder cancer screening.

Our study evaluated the BTA (bladder tumor antigen) stat test kit as a primary screening device for the detection of transitional-cell carcinoma (TCC) of the bladder, with direct comparison by voided urine cytology (VUC) on the same specimens. The unfixed voided urine of 100 patients with no history of bladder cancer who had signs and symptoms of dysuria, incontinence, and gross hematuria and microhematuria were tested using the one-step BTA stat test kit before processing via the cytospin technique for fluid cytological evaluation. The patients in the study were followed for up to 12 mo with repeated urine cytological testing, cystoscopy, and bladder biopsy when clinically indicated. Nineteen cases tested positive, and 81 cases tested negative on the BTA stat test. VUC diagnosed three cases as unequivocally positive for TCC, 93 cases as negative, and four cases in which unqualified atypical urothelial cells were noted. TCC was confirmed by cystoscopy and bladder biopsy in three of three cases diagnosed by VUC and in three of 19 cases that tested positive by the BTA stat test. These findings resulted in an 84% false-positive rate for the BTA stat test and no false-positive cases for VUC during the 12-mo follow-up period. The results indicate that the sensitivity and specificity of BTA stat test are comparable to those of VUC; however, owing to a relatively high false-positive rate, it can at best act as an adjunct to urine cytological study for bladder cancer screening.     

The Utility of Fluorescence In Situ Hybridization for Detection of Bladder Urothelial Carcinoma in Routine Clinical Practice

To evaluate the ability of fluorescence in situ hybridization (FISH) in detecting bladder urothelial carcinoma (BUC), FISH and cytology were compared for the evaluation of 308 consecutive urine samples from patients suspected of having BUC. All patients underwent cystoscopy for identification of bladder lesions. The FISH results were compared with the cytology assessment. In all, 122 patients had confirmed BUC. Among them, 68 (55.7%) were FISH-positive, while only 33 (27%) were positive on cytology. According to disease stage (superficial vs. invasive) and grade (low vs. high), the sensitivities of FISH were also significantly higher than those of cytology in all categories. Moreover, in 36 patients who had no visible tumor with flat, erythematous mucosa (suspicious lesion), FISH was more sensitive than cytology for the detection of BUC (83.3% vs. 33.3%, P=0.002). The FISH was negative in 168 (90.3%) of 186 patients with no histological evidence of BUC or negative cystoscopy findings. The sensitivity of FISH for detecting BUC was superior to that of cytology, regardless of tumor stage and grade. FISH is a significant additional and complementary method for detection of BUC in patients who have suspicious lesions on cystoscopy.     

DNA flow cytometry and bladder irrigation cytology in detection of bladder carcinoma

In this study we assessed the role of DNA flow cytometry (FCM) as an adjunct to bladder irrigation cytology to detect carcinoma of the bladder. We selected only those cases who had urinary symptoms and cystoscopic examination or histology-proven cases of bladder cancer who underwent cystoscopy for a follow-up study. Cystoscopy, cytologic examination, and DNA FCM were performed in every case. There were 9 fresh cases and 21 follow-up cases of proven transitional-cell carcinoma (TCC) of the bladder. Cystoscopy revealed growth in all 9 fresh cases as well as in 11 follow-up cases. Cytology was positive in 16 cases, out of which there were 8 each of fresh and recurrent cases. None of the cases showed positive cytology with negative cystoscopy findings. DNA FCM was positive in 13 cases. Aneuploidy was detected in 5 cases, out of which there were 3 hyperdiploid and 2 hypodiploid cases. Nine cases had high (equal or more than 10%) S and G2-M phase cells, ranging from 10-19.36%. One case showed aneuploidy along with high S-G2M phase. Both cytology and DNA FCM were positive in 9 cases. In 2 cases, DNA FCM showed aneuploidy, but cytology and cystoscopy were negative. The sensitivity and specificity of the bladder wash cytology were 80% and 100%, and those for DNA FCM were 55% and 83.3%, respectively. We conclude that both bladder wash cytology and DNA FCM techniques should be done in all the cases of suspected TCC to detect more number of positive cases.     

Urinary nuclear matrix protein 22 (NMP22): a diagnostic adjunct to urine cytologic examination for the detection of recurrent transitional-cell carcinoma of the bladder.

This study compares urine nuclear matrix protein 22 (NMP22) immunoassay and conventional urine cytologic examination for detecting recurrent transitional-cell carcinoma (TCC) of the urinary bladder. One hundred twenty-eight urine specimens from 107 patients with a history of TCC of the urinary bladder were studied. NMP22 immunoassay and conventional cytologic examination were performed on each specimen. The NMP22 and cytology results were then compared with the results of subsequent cystoscopies/surgical biopsies performed over a 6-mo follow-up period. The sensitivity of urine cytologic study for predicting recurrent TCC was 60%, while the sensitivity of NMP22 assay was 47%. When both NMP22 assay results and the cytologic interpretation were positive for TCC, the positive predictive value of the combined tests was 74%. When both tests showed negative results, the negative predictive power was 81%. Our findings suggest that urine NMP22 assay may represent a useful diagnostic adjunct to conventional urine cytologic examination for the detection of recurrent TCC of the urinary bladder.     

Evaluation of the NMP22 test and comparison with voided urine cytology in the detection of bladder cancer

The purpose of this study was to assess the clinical performance of the NMP22 test and to compare it with that of voided urine cytology for the detection of bladder cancer. The NMP22 test was evaluated in two groups of patients. The first group was comprised of patients with histologically confirmed active transitional cell carcinoma (TCC) of the bladder, and the second group contained those with a history of bladder TCC but that were considered to have no evidence of disease on the basis of cystoscopic evaluation of bladder and/or biopsy. Sensitivity was determined in voided urine samples from patients with active TCC of the bladder. Specificity was determined in the urine samples of patients with a history of bladder TCC but no current evidence of disease. The NMP22 test was positive in 53 of 70 samples from patients with active bladder TCC. The sensitivity of the NMP22 test (75.7%) is significantly better than that of voided urine cytology (55.7%). The specificity of the NMP22 test and of voided urine cytology were 72.2% and 88.9% respectively, in patients with a history of bladder TCC but no current evidence of disease. There was no significant difference between the specificity of NMP22 and that of urine cytology. The NMP22 test is superior to voided urine cytology in the detection of TCC of the bladder. The results of this study indicate that the NMP22 test is an useful adjunct to cystoscopy in the detection and monitoring of TCC of the bladder.     

Multiprobe FISH for enhanced detection of bladder cancer in voided urine specimens and bladder washings.

The aim of this study was to evaluate the UroVysion (Vysis, Downers Grove, IL) fluorescence in situ hybridization (FISH) test for improved detection of bladder cancer in urinary specimens. Three groups of specimens were examined, including voided urine specimens (1) collected before resection of bladder cancer, (2) from cystoscopically negative bladders of patients with previous bladder cancer, and (3) from patients with benign prostatic hyperplasia (controls). FISH positivity was defined as more than 2 urothelial cells with an abnormal signal copy number of at least 1 of the 4 probes. FISH was positive in 1 of 27 control specimens and in 33 (73%) of 45 pTa, 12 (100%) of 12 pT1, and 13 (100%) of 13 pT2-4 tumors. The results were similar in a series of 68 bladder washings. In addition, FISH of voided urine specimens was positive in 5 of 10 patients with negative follow-up cystoscopy results. Subsequent recurrence was found in 4 of these patients but in none of 5 patients with FISH-negative results. Multiprobe FISH markedly improves the sensitivity and specificity of cytology for the detection of bladder cancer in urine specimens.     

Follicular cystitis in urine cytology: A clinical and cytopathologic study

Despite being an important differential diagnosis of bladder tumor on cystoscopy, follicular cystitis (FC) is rarely diagnosed on cytologic material. We performed a retrospective study on cases of FC diagnosed on bladder biopsy and/or urine cytology in our institution. A total of 35 cases of FC were identified with a female predominance (F:M = 2:1). Hematuria was the most common clinical presentation. Cystoscopic findings included mass lesions, yellow plaques, and surface erythema. History of urinary tract infection was reported in 48% of the patients, and majority of those patients had positive concurrent urine culture, most commonly with beta‐hemolytic streptococcus, Group B. A total of 17 out of 35 patients had urine cytology specimens. When the presence of follicular dendritic cells in clusters of variously sized lymphocytes is used as the cytological diagnostic criterion, 6 out of 17 cases were diagnosed as FC and 5 out of 6 were confirmed by concurrent biopsy. This retrospective study not only analyzed the clinical characteristics of FC but also elucidated the cytological diagnostic criteria of FC and confirmed its specificity.     

Routine transurethral biopsy of the bladder is not necessary to evaluate the response to bacillus calmette-guerin therapy

We evaluated the need for transurethral biopsy at first follow-up after intravesical bacillus Calmette-Guerin (BCG) therapy for superficial bladder cancer. The records of 84 patients with superficial bladder cancer who received a 6- or 8-week course of BCG were reviewed. Pathological results before BCG, cystoscopic findings, urinary cytology, and biopsy results for evaluation of BCG therapy were reviewed. All 19 patients with positive urinary cytology had evidence of positive bladder biopsy results. Fifty-three of 54 patients (98.1%) with no visible recurrent tumor and negative urinary cytology demonstrated negative pathological results on bladder biopsy. When not found in conjunction with positive urinary cytology, erythematous mucosa on cystoscopy was not an indicator of tumor recurrence or residual cancer. In conclusion, routine transurethral biopsy of the bladder for evaluating the response to BCG intravesical therapy is not necessary in patients who have no visible tumor on cystoscopy and negative urinary cytology.     

Usefulness of NMP22 as an adjunct to a typical urine cytology and low‐grade urothelial carcinoma

The usefulness of urine cytology combined with NMP22 was evaluated for the primary diagnosis of urothelial carcinoma. Of 53 clinically suspected patients, histopathological diagnoses were low‐grade urothelial carcinoma (25), high‐grade urothelial carcinoma (13), and inflammatory lesions (15). Cytology was positive in 25 and negative in 14 patients. Fourteen of 25 low‐grade urothelial carcinoma and 11/13 high‐grade urothelial carcinoma were diagnosed correctly on urine cytology. Atypical cells seen in 14 patients were categorized as inconclusive for malignancy. The overall sensitivity of urine cytology was 65.8%, whereas specificity was 100%. NMP22 was positive in 33 patients. Of these 30, 18 low‐grade and 12 high‐grade lesions were true positive. Of the 20 NMP22, eight negative cases were false‐negative. Ten of 15 with negative histopathology were also negative for NMP22, three were false‐positive, and two showed erratic results. Nine of 14 cases with atypical urine cytology were positive for NMP22. Eight of these showed low‐grade carcinoma on histopathology. The sensitivity of BladderChek NMP22 test was 79%, whereas specificity was 80%. NMP22 BladderChek test is a useful adjunct to urine cytology in atypical and low‐grade carcinoma. Diagn. Cytopathol. 2010;38:788–790. © 2009 Wiley‐Liss, Inc.     

Fascin stain as a potential marker of invasiveness in carcinomas of the urinary bladder: a retrospective study with biopsy and cytology correlation

The evaluation of invasion in urothelial carcinomas of the urinary bladder cannot be determined on cytology and can be particularly challenging in biopsy cases with limited sampling. Recent studies of bladder resection specimens suggest that fascin overexpression may be a marker of aggressive urothelial carcinomas and can help facilitate the assessment of invasion. In this study, we evaluated urine cytology and corresponding biopsy specimens with proven invasive urothelial carcinoma for fascin expression by immunohistochemistry. Thirty‐five patients diagnosed with positive urine cytology and biopsy‐proven invasive urothelial carcinoma between January 2003 and February 2009 were identified. We found increased fascin expression in 100% (35/35) of SurePathTM&!trade; urine cytology preparations as well as 100% (35/35) of corresponding biopsy cases with invasive urothelial carcinoma. On urine cytology, cytoplasmic fascin staining was moderate to intense in malignant tumor cell clusters and single cells and not observed in benign urothelial cells. Staining in biopsy cases was generally intense and cytoplasmic and present in both the invasive (100%) and noninvasive (31%) components of the lesion. These findings uphold the association of increased fascin expression in invasive urothelial carcinomas of the urinary bladder. We furthermore demonstrate that fascin staining can be performed successfully on SurePathTM&!trade; urine cytology preparations in which increased fascin expression correlates with invasion on biopsy. While not a definitive marker of invasion, as it is observed in in situ carcinoma, we conclude that the utilization of fascin immunohistochemistry on urine cytology might serve as a useful adjunct in predicting invasiveness in subsequent biopsies. Diagn. Cytopathol. 2010. © 2010 Wiley‐Liss, Inc.     

Cyclooxygenase‐2 expression on urothelial and inflammatory cells of cystoscopic biopsies and urine cytology as a possible predictive marker for bladder carcinoma

Cyclooxygenase‐2 (COX‐2) is a key inducible enzyme involved in the production of prostaglandins. It contributes to human carcinogenesis by various mechanisms. The aim of the current study was to elucidate the possible involvement of COX‐2 in human bladder carcinoma by examining its expression on both urothelial and inflammatory cells in tissue biopsies and urine cytology samples of different urinary bladder lesions. A total of 65 patients were included in the study and were selected from cases admitted to Urology Department, Theodor Bilharz Research Institute (TBRI), Giza, Egypt. They represented seven control cases with almost normal‐looking bladder tissue; pure chronic cystitis (n=12); premalignant lesions (18) in the form of squamous metaplasia (n=8) or urothelial dysplasia (n=10) as well as transitional cell carcinoma (TCC) (n=18), and squamous cell carcinoma (SqCC) (n=10). Immunohistochemistry of formalin‐fixed, paraffin‐embedded tissue sections and urine cytology samples was performed for all cases using COX‐2 (H‐62): sc‐7951, a rabbit polyclonal antibody. The study revealed positive COX‐2 expression on the urothelial and inflammatory cells of cystoscopic biopsies from all cases of pure chronic cystitis, squamous metaplasia and SqCC compared with 42.8% and 71.4% of normal controls, respectively. The score of urothelial COX‐2 expression was sequentially up‐regulated from normal to chronic cystitis (either pure or associated with premalignant changes) (p<0.05) to malignant changes (p<0.05). However, the inflammatory cellular expression was down‐regulated with malignant transformation compared with chronic cystitis (p<0.05). In TCC, COX‐2 was over‐expressed on both urothelial and inflammatory cells in advanced tumors. Urine cytology samples were positive for COX‐2 in a comparable manner to that observed in cystoscopic biopsies. Accordingly, the results of the current study have provided new information in two aspects: First, is the possibility of using the differential COX‐2 expression on both inflammatory and urothelial cells as markers for premalignant or malignant transformation; second, besides cystoscopy, urine cytology was found to have a high sensitivity for COX‐2 expression and hence proved to be valuable in malignancy as a non‐invasive substitute for cystoscopy.     

Cytokeratin-20 immunocytochemistry in voided urine cytology and its comparison with nuclear matrix protein-22 and urine cytology in the detection of urothelial carcinoma

This study was done on 59 subjects (42 urinary bladder carcinoma patients and 17 non‐neoplastic controls). Urine cytology and bladder chek NMP22 test was done on all cases. CK20 immunostaining was performed on archived papanicolaou stained urine cytology smears in 34 cases (27 bladder carcinoma and 7 negative controls). Results of all three tests (cytology, NMP22, and CK20 immunostaining) were compared with histopathology to evaluate the accuracy of individual test. The combination of cytology and NMP22 was compared with combination of cytology and CK20 immunostaining for detection of bladder carcinoma. NMP22 had sensitivity of 92.9% and specificity of 70.6%, as compared with voided urine cytology (sensitivity of 76.2% and specificity of 76.5%) and CK20 immunostaining (sensitivity of 70.4% and specificity of 71.4%). Combination of cytology and NMP22 gave better results (sensitivity of 88.1% and specificity of 88.2%) than combination of cytology and CK20 immunostaining or any other test in isolation. Diagn. Cytopathol. 2012. © 2011 Wiley Periodicals, Inc.     

Detection of bladder cancer by semi-automated microsatellite analysis of urine sediment

Patients with transitional-cell carcinoma (TCC) require careful follow-up due to the high risk of recurrence. Cystoscopy and biopsy are reliable but invasive, while urine cytology is plagued by low sensitivity. It has recently been shown that allelic abnormalities detected by microsatellite analysis of DNA extracted from urine can be used to diagnose TCC with high reliability. As this analysis by classic techniques is unfeasible in a clinical setting, we performed a pilot study to determine the possibility of applying quick DNA extraction methods with laser detection and computer-based analysis of 15 fluorescently labeled PCR amplified microsatellites to detect molecular anomalies in urine sediment in 25 TCC follow-up patients. Of the eighteen cases with recurrent TCC, 14 (78%) were positive by the molecular test whereas only eight (44%) were detected by cytology. Of the seven patients with negative cystoscopy, one resulted positive by the molecular test and had recurrent TCC six-months later. Thus, this microsatellite analysis correctly predicted the clinical diagnosis in 84% (21/25) of cases, compared to 60% by cytology. The application of these semi-automated procedures allows the analysis of 18 samples with 15 markers in one day, encouraging a more expedient introduction into routine clinical use.     

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.     

Detection of bladder tumor by urine cytology in cases of prostatic hypertrophy

Routine urine cytology has been performed for 809 male patients presenting with symptoms due to prostatic hypertrophy. In 6.42% of the cases, bladder tumor was revealed incidentally diagnosed by urine cytology. Cystoscopy and bladder biopsies were performed to confirm malignancy. The age of these patients ranged from 48–84 years (mean 65.2) and they complained mainly of irritative bladder symptoms. It is therefore strongly indicated that all patients with prostatic disease should have routine cytological examination of urine sediments. Diagn Cytopathol 1996;15:409–411     

微小膀胱副神经节瘤的诊治分析

目的探讨微小膀胱副神经节瘤(micro-PUB)(1 cm)的临床诊治特点。方法回顾性分析北京协和医院2005年1月至2015年1月收治的5例微小PUB患者的临床资料。男性2例,女性3例,年龄41~65岁,平均51岁。4例患者为功能性肿瘤,1例为非功能性肿瘤。各项检查对PUB肿瘤的检出率分别如下:作为定性诊断作用的24 h尿儿茶酚胺(CA)检查的阳性率为75%(3/4),作为定位诊断作用的超声、CT、MRI、奥曲肽显像及131I-间碘苄胍(MIBG)检查的阳性率分别是80%(4/5)、20%(1/5)、75%(3/4)、25%(1/4)及33%(1/3)。所有患者均行经尿道手术切除肿瘤,术中肿瘤定位困难时可通过过度充盈膀胱或超声定位方法确定肿瘤位置。结果所有患者术中均顺利找到并切除肿瘤,无中转开放,术中寻找肿瘤时间为0.5~26 min,切除时间3~10 min,术后病理为PUB。随访6个月~8年,排尿后高血压症状消失,复查24 h CA、泌尿超声、膀胱镜及全身MRI,未见肿瘤复发及远处转移。结论超声和MRI对于微小PUB的发现率较CT高,应作为首选检查。治疗微小PUB应采用经尿道手术,术中如无法发现肿瘤,可使膀胱过度充盈和超声定位确定肿瘤位置。     

Koilocytosis: Correlations with high‐risk HPV and its comparison on tissue sections and cytology,urothelial carcinoma

The aim of this study were (1) To correlate koilocytosis with high risk HPV(HrHPV) DNA in urinary bladder carcinoma and (2) To compare detection of koilocytosis on tissue sections and urine cytology. Biopsy and cytologic specimens from 33 patients of urinary bladder carcinoma were analyzed. HPV DNA was detected by PCR on biopsy specimens using consensus primers MY09 and MY11. Koilocytosis was assessed both on tissue sections and urine cytology. HrHPV DNA was found in 14 of 33 bladder carcinoma. Koilocytosis was seen in tissue sections from 13 patients. Eleven of these were HrHPV DNA positive (positive predictive value 84.6%). Koilocytosis was seen in urine cytology in three patients. All three were positive for HrHPV DNA. To conclude koilocytosis is a good morphological marker for HrHPV DNA in the urothelium. Tissue sections are better than cytologic smears for detection of koilocytes. Diagn. Cytopathol. 2009. © 2009 Wiley‐Liss, Inc.