荧光原位杂交在膀胱尿路上皮癌诊断中的应用

目的评价尿脱落细胞荧光原位交(fluorescenceinsituhybridization,FISH)检测在膀胱肿瘤诊断中的应用价值。方法分别对69例疑似膀胱尿路上皮癌及20例对照组的尿液标本进行FISH及细胞学检测,比较两者诊断的敏感性及特异性,统计膀胱尿路上皮癌各个染色体畸变的几率。结果 FISH诊断膀胱尿路上皮癌的总的敏感度高于尿脱落细胞学检查(分别为79.7%、22.0%,P<0.05),两者的特异度分别为93.3%、100%(P>0.05)。结论 FISH在诊断膀胱尿路上皮癌中敏感性高于尿细胞学检查,同时其特异性亦较高,在早期诊断中具有重要意义。     

荧光原位杂交技术在尿路上皮癌诊断中的应用

目的 探讨荧光原位杂交技术( FISH)在尿路上皮癌诊断中的应用价值.方法 采用FISH检测100例血尿患者尿脱落细胞中第3、7、17号染色体和第9号染色体p16位点异常,以组织病理学确诊尿路上皮癌为金标准,评估FISH诊断的敏感度和特异度,并与尿细胞学检查结果进行比较.结果 FISH检测和尿细胞学检查诊断尿路上皮癌的敏感度分别为82.5％和49.2％, 差异有统计学意义(P＜0.05)；特异度分别为86.7％和96.6％,差异无统计学意义(P＞0.05).结论 与尿细胞学比较,FISH诊断尿路上皮癌具有较高的敏感度和相似的特异度.     

荧光原位杂交技术在血尿患者膀胱尿路上皮癌筛查中的应用

目的 评价荧光原位杂交(fluorescence in situ hybridization,FISH)技术在血尿患者膀胱尿路上皮癌筛查中的临床应用价值,为从尿液脱落细胞中早期诊断膀胱癌提供新的途径.方法 选择20例健康正常人,通过FISH技术检测9号染色体p16位点、3号染色体、7号染色体及17号染色体变异情况,建立阈值.100例血尿患者留取晨尿,分别进行尿脱落细胞学检查和FISH技术检查.统计分析FISH方法的敏感度和特异度.结果 探针检测p16基因完全缺失阈值为3.9%,部分缺失为5.4%,扩增为3.2%;3号染色体缺失阈值为4.0%.扩增为3.5%;7号染色体缺失阈值为4.0%,扩增为2.8%;17号染色体缺失阈值为5.8%,扩增为3.4%.以至少两种探针检测结果超过阈值或一种探针检测结果至少存在两种异常为诊断阳性.FISH技术在血尿患者膀胱尿路上皮癌筛查中的敏感度和特异度分别为93.6%和100%,尿脱落细胞学检测分别为46.8%和100%.结论 FISH技术无创快速,对血尿患者膀胱尿路上皮癌筛查的敏感度优于尿脱落细胞学检查方法.     

尿脱落细胞FISH检测在恶性血尿病因诊断中的应用

目的：比较尿脱落细胞荧光原位杂交（Fluorescence in situ hybridization，FISH）检测、细胞学分析和膀胱镜对恶性血尿诊断的临床有效性及应用价值，为建立或完善恶性血尿病因诊断提供理论基础。方法：10例正常人通过FISH技术检测9号染色体p16位点、3号染色体、7号染色体及17号染色体的变异情况，建立FISH检测尿脱落细胞的方法学平台及标准检测方法同时确立阈值。同时收集100例血尿患者晨尿同步进行细胞学分析、膀胱镜检查及FISH检测。以组织病理确诊为膀胱尿路上皮癌为金标准，评估FISH诊断的特异度和敏感度及与膀胱癌发生及发展的关系。结果：FISH诊断尿路上皮癌的特异度为86．1％，敏感度为89．1％；尿细胞学诊断的特异度为97．2％，敏感度为23．4％。两者相比，敏感度差异有统计学意义（P〈0．05），而特异度差异无统计学意义（P〉0．05）。结论：与尿细胞学相比，FISH在恶性血尿病因的诊断中具有较高的灵敏度和相似的特异度，同时无创快速，可作为筛查和诊断尿路上皮癌的新方法。     

荧光原位杂交技术诊断和监测膀胱尿路上皮癌复发的临床应用

目的 探讨荧光原位杂交技术( fluorescence in situ hybridization,FISH)诊断和监测膀胱尿路上皮癌复发的可行性,并与尿脱落细胞学进行对比.方法 应用荧光标记的3、7、17号染色体着丝粒探针和9号染色体p16位点9p21区带探针,对80例疑似膀胱肿瘤患者尿脱落细胞进行FISH检测,并同期行尿脱落细胞学检测,以病理结果作为诊断“金标准”,和FISH进行比较;以20例健康志愿者作为正常对照组,建立FISH阈值.结果 FISH和尿脱落细胞学诊断膀胱尿路上皮癌特异度分别为86.7％和100％,差异无统计学意义(P＞0.05).FISH和尿脱落细胞学诊断膀胱尿路上皮癌总体灵敏度分别为80％和32.3％.FISH和尿脱落细胞学在灵敏度以及肿瘤分期、分级、初发和复发等方面差异均有统计学意义(P＜0.05).结论 FISH可能成为一种新的早期诊断和监测膀胱癌术后复发的有效方法.     

荧光原位杂交法在膀胱尿路上皮癌诊断中的应用

目的：探讨荧光原位杂交法（FISH）在膀胱尿路上皮癌诊断中的应用。方法：选取20例非尿路上皮癌和40例膀胱尿路上皮癌的人群尿液作常规尿脱落细胞学检查和FISH检测。结果：FISH技术的敏感性为82．5％,显著高于常规尿脱落细胞学的敏感性25．0％（P〈0．05）;FISH技术和常规脱落尿细胞学检查的特异性均为100％,两者在特异性方面差异无统计学意义（P〉0．05）。结论：荧光原位杂交法在膀胱尿路上皮癌诊断中的特异性与常规尿脱落细胞学检查一致,但其敏感性显著高于常规尿脱落细胞学检查,所以,FISH技术更有望成为膀胱尿路上皮癌无创性的诊断和检测手段。     

荧光原位杂交技术在膀胱尿路上皮癌诊断中的临床应用

目的 探讨荧光原位杂交(FISH)技术运用于膀胱尿路上皮癌的诊断价值.方法 收集20例健康志愿者的新鲜晨尿,运用荧光标记的3号、7号、17号染色体着丝粒探针及9号染色体p16位点探针,对尿液标本中的脱落细胞染色体进行FISH技术检测,建立正常人群的阈值.收集158例怀疑为膀胱尿路上皮癌患者的新鲜晨尿,在行膀胱镜检查前,同期进行FISH技术与尿脱落细胞学检测,运用统计学方法,比较FISH技术与尿脱落细胞学检测的敏感性与特异性.结果 FISH与尿脱落细胞学的敏感性分别为84.8%和43.8%,FISH敏感性高于尿脱落细胞(P<0.05),FISH与尿脱落细胞学特异性分别为89.1%和87.0%,两者无统计学差异(P>0.05),在不同的肿瘤病理分级中,FISH的敏感性都高于尿脱落细胞,并且FISH敏感性随肿瘤分级逐级升高(P<0.05).结论 FISH技术具有较高的敏感性和特异性,可以作为国人膀胱尿路上皮癌筛查、诊断的新方法.     

荧光原位杂交技术在膀胱尿路上皮癌诊断中的应用价值

目的 探讨荧光原位杂交技术(fluorescence in situ hybridization,FISH)在膀胱尿路上皮癌诊断中的应用价值. 方法 收集81例膀胱尿路上皮癌患者的新鲜尿液标本,分别行FISH和尿细胞学检测,并行膀胱镜检查;病理诊断泌尿系良性疾病12例合并8例临床诊断泌尿系良性疾病患者的尿液标本作对照.统计学分析FISH和尿细胞学检测诊断的特征值. 结果 81例均经病理检查确诊膀胱尿路上皮癌.非肌层浸润性膀胱尿路上皮癌34例,肌层浸润性癌14例;低级别42例,高级别24例;由于送检组织不完整,无法准确分期、分级者分别为33例和15例.FISH、尿细胞学和膀胱镜检查诊断膀胱尿路上皮癌的敏感性分别为72.8％ (59/81) 、27.2％ (22/81)和97.5％ (79/81),FISH诊断敏感性高于尿细胞学检测(P＜0.05),但低于膀胱镜检查(P ＜0.05);FISH和尿细胞学检测诊断特异性分别为85.0％(17/20)、100.0％ (20/20),二者比较差异无统计学意义(P＞0.05).FISH和尿细胞学检测诊断符合率分别为75.2％ (76/101)、41.6％(42/101),二者比较差异有统计学意义(P＜0.05). 结论 FISH检测诊断膀胱尿路上皮癌敏感性高、特异性强、无创,临床应用价值高.     

荧光原位杂交技术在血尿待诊病因诊断中的临床应用分析

目的:评价非侵袭性尿脱落细胞学和荧光原位杂交技术(fluorescence in situ hybridzation,FISH)在血尿待诊病因诊断中的应用价值,为建立和完善血尿待诊病因诊断提供理论依据。方法:选取2011年8月~2013年12月因血尿体检的正常人10例,通过FISH技术检测9号染色体p16号位点、3号染色体、7号染色体、17号染色体,建立FISH检测方法并确立阈值。同时收集185例血尿患者晨尿同步进行尿脱落细胞学分析及FISH检测。以病理诊断尿路上皮癌为金标准,评估FISH诊断的特异性和敏感性及与膀胱癌发生和发展的关系。结果:FISH检测尿路上皮癌患者的特异性为91.8%,敏感度为94.3%;尿脱落细胞学诊断的特异性为97.3%,敏感性为36.1%。两者相比,敏感性差异有统计学意义(P0.05),而特异性差异无统计学意义。结论:与尿脱落细胞学相比,FISH在血尿病因学膀胱癌患者的诊断中具有较高灵敏度,而特异度两者相近。FISH使低级别浅表型尿路上皮癌的准确率明显提高,几乎能检测出所有高级别的浸润性膀胱癌。FISH的染色体3、7、9和17是血尿恶性疾病鉴别诊断尿路上皮癌的一种有前途、无创性分子检测方法。     

不同标准对荧光原位杂交技术在膀胱癌诊断中应用的影响

目的：探讨不同标准对荧光原位杂交技术（fluorescence in situ hybridization,FISH）诊断膀胱癌的敏感度和特异性的影响。方法：选择20例健康人为正常组,计算FISH检查正常阈值;选择143例血尿患者为病例组．经F1SH检查、尿脱落细胞学检查,比较Urovysion膀胱癌探针斌剂盒标准和正常闯值标准诊断膀胱癌的敏感度和特异性。结果：采用Urovysion标准和正常阈值标准对膀胱癌的诊断敏感度分别为73．1％和100％．均较尿脱落细胞学检查明显增高,三者对膀胱癌诊断的特异性分别为90．0％、86%和100％。Urovysion标准与尿脱落细胞学联合检查时对膀胱癌诊断的敏感度明显升高,差异具有统计学意义（P〈0．01）。结论：FISH检查较尿脱落细胞学检查诊断膀胱癌的敏感度显著提高,相比Urovysion际准,正常闯值更适合FISH诊断膀胱癌的标准。FISH与尿脱落细胞学联合检查能显著提高膀胱癌的诊断敏感度。     

Clinical utility of a multiprobe FISH assay in voided urine specimens for the detection of bladder cancer and its recurrences,compared with urinary cytology

OBJECTIVES: To evaluate the clinical utility of a Multi-color FISH (fluorescence in situ hybridization) assay in voided urine specimens for the detection of bladder cancer and its recurrences, comparing the results with those afforded by urinary cytology. METHODS: Voided urine samples from 86 patients were obtained for urine cytology and FISH analysis. The latter was performed using a mixture of fluorescent labeled DNA probes for the centromeric regions of chromosomes 3, 7 and 17, and the 9p21 region. Cystoscopy with biopsy or tumor resection was performed in all patients, comparing the pathological results with the cytological and FISH findings. RESULTS: Urinary cytology affords an overall sensitivity of 63.8%, the figure being 25% for grade 1, 66.6% for grade 2 and 94.7% for grade 3 tumors. The sensitivities for FISH were 53.3% for grade 1, 83.3% for grade 2 and 100% for grade 3 tumors, with an overall sensitivity of 80.4%. The specificities of urinary cytology and FISH were 86.1 and 85.3%, respectively. CONCLUSIONS: FISH improves the sensitivity rates obtained with urine cytology for bladder cancer detection in all tumor grades and stages, and offers similar specificity. FISH doubles the accuracy of urinary cytology in application to low grade-stage tumors, and detects all high grade infiltrating tumors.     

Multitarget fluorescence in situ hybridization assay detects transitional cell carcinoma in the majority of patients with bladder cancer and atypical or negative urine cytology

PURPOSE: The multitarget fluorescence in situ hybridization (FISH) probe set UroVysion (Vysis, Downers Grove, Illinois), containing probes to chromosomes 3, 7 and 17, and to the 9p21 band, has been recently shown to have high sensitivity and specificity for detecting transitional cell carcinoma. In this study we retrospectively tested 120 urine samples from patients with atypical, suspicious and negative cytology for whom concurrent and followup bladder biopsy data were available. We evaluated the ability of FISH to identify malignant cells in cytologically equivocal or negative cases. MATERIALS AND METHODS: Archived slides from 120 voided (47) or instrumented (73) urine cytology specimens from patients with concurrent bladder biopsy and a minimum of 12 months of biopsy followup were subjected to hybridization with UroVysion. The cohort included patients with biopsy proven transitional cell carcinoma, which was grades 1 to 3 in 23, 35 and 24, respectively, and stages pTis in 3, pTa in 64, pT1 in 6, pT2 in 6 and pT4 in 3, while it showed negative histology in 38. Cytology findings were suspicious, atypical and negative for transitional cell carcinoma in 31, 49 and 40 cases, respectively. A positive FISH result was defined as 5 transitional cells or greater with a gain of 2 or more of chromosomes 3, 7 or 17, 12 cells or greater with 9p21 deletion, or 10% or greater of cells with isolated trisomy of 1 of chromosomes 3, 7 and 17. RESULTS: All except 12 of the 82 biopsy proven transitional cell carcinoma cases (11 pTa and 1 pT1 tumors) were positive by FISH (85% sensitivity). Sensitivity in patients with suspicious, atypical and negative cytology was 100%, 89% and 60%, respectively. Nine patients with atypical cytology had positive FISH in the setting of a negative concurrent bladder biopsy. However, 8 of these 9 patients (89%) had biopsy proven transitional cell carcinoma within 12 months following the date when the sample tested by FISH was obtained. The last of these patients with false-positive results had previously documented pTis disease, which was also present in the next bladder biopsy 15 months following the positive FISH result. The remaining 29 specimens from patients with negative biopsy and a negative 12-month followup tested negative by FISH (97% overall specificity). CONCLUSIONS: The UroVysion FISH assay provides high sensitivity and specificity to detect transitional cell carcinoma in cytologically equivocal and negative urine samples. These results emphasize the important role of this assay in the management of bladder cancer.     

The UroVysion fluorescence in situ hybridization assay is an effective tool for monitoring recurrence of bladder cancer

The newly developed UroVysion fluorescence in situ hybridization (FISH) probe was applied to urine specimens from 19 patients being monitored for recurrence of bladder cancer. The results for the multi-target DNA FISH assay were compared with independent analyses of urine cytology and flexible cystoscopy. Patients with tumors identified through the cystoscopy exam were biopsied and/or underwent surgery. In 12 patients with normal cytoscopy, cytology and FISH were also normal. Therefore, the specificity of these two tests was 100%. In 7 patients, a tumor was diagnosed by cystoscopy, and 3 of them had abnormal urine cytology while 6 of them had an abnormal result in the FISH assay. Accordingly, the sensitivity was 43% for the cytology and 87% for the FISH test. Interestingly, a pT1G3 tumor in a bladder diverticulum was not detected by cytology or the FISH test. These results agreed with a large series previously published using similar FISH probes and support the proposal for a multicenter trial to confirm the usefulness of the UroVysion probe as a screening tool to select patients for cystoscopy.     

The simultaneous use of telomerase, cytokeratin 20 and CD4 for bladder cancer detection in urine

OBJECTIVE: Because of the low sensitivity of urinary cytological diagnosis of urinary bladder carcinoma, new molecular diagnostic methods have been proposed. We decided to verify the expression of telomerase mRNA coding for the catalytic component (hTRT), cytokeratin 20 (CK20) and CD4 antigen mRNAs in urine as possible diagnostic tool. METHODS: Evaluation of hTRT, CK20, CD4 mRNAs was performed in 50 ml of naturally voided urine of 205 patients of which 153 with bladder cancer (Tis, n = 11; TaGx, n = 4; TaG1, n = 25; TaG2, n = 26; TaG3, n = 8; T1G1, n = 16; T1G2, n = 17; T1G3, n = 20; T2G2, n = 6; T2G3, n = 13; T3G3, n = 7) and 52 controls. A quantitative expression of hTRT at mRNA level versus TRAP (telomeric repeat amplification protocol) assay was performed in 20 patients and 14 controls. The expression of RT-PCR for hTRT, CK20, CD4 versus urinary cytology was analysed in 44 patients with bladder cancer. Evaluating the three molecular markers together, the result was considered correct when at least two of the markers were positive, suspected when only one marker was positive and negative for diagnosis of tumour when all markers were negative. The performance of the diagnostic model resulted from the logistic analysis evaluated with receiver operating characteristics (ROC) curve analysis. RESULTS: The sensitivity detected for each tumour marker was as follows: for hTRT 90.8%, for CK20 84.3% and for CD4 was 64.7%, while the specificity was 94.2% for CD4 and 78.8% for both hTRT and CK20. When a simultaneous evaluation of the three tumour markers was considered, 88.2% of the diagnoses were correct, 11.8% were suspected for tumour and none were mistaken. When compared with cytology, the simultaneous use of the three markers allowed reaching a correct diagnosis in 88% of the cases in comparison to 25% by urinary cytology. The sensitivity in the detection of bladder cancer was higher for hTRT at mRNA level in comparison with the enzymatic activity detection with TRAP (90% vs. 35%) while the specificity for both markers resulted very high (100%). CONCLUSIONS: These data show that in the future the diagnostic improvement of urine based molecular markers for the detection of bladder cancer in the urine could improve the sensitivity of urinary cytology reducing the need of a cystoscopy.     

荧光原位杂交法和全自动图像细胞仪在膀胱尿路上皮癌诊断中的应用

目的 探讨荧光原位杂交法(FISH)和全自动图像细胞仪(ICM)在膀胱尿路上皮癌诊断中的应用.方法 在2008年8月至2009年3月共选取60例患者,包括20例非尿路上皮癌和40例膀胱尿路上皮癌的患者,取患者的尿液作常规尿细胞学检查、FISH和ICM检测.结果 FISH的敏感性显著高于ICM的敏感性(82.5%比62.5%,P＜0.05)和常规尿细胞学的敏感性(82.5%比25.0%,P＜0.05),同时ICM敏感性也高于常规尿细胞学的敏感性(62.5%比25.0%,P＜0.05);FISH、ICM和常规尿细胞学检查的特异性都为100%,三者在特异性方面差异无统计学意义(P＞0.05).FISH、ICM和常规尿脱落细胞学检测的敏感性与病理分期无相关性(P＞0.05),但与分级有相关性(P＜0.05).结论 FISH和ICM在膀胱尿路上皮癌诊断中,其特异性和常规尿细胞学检查一致,但敏感性显著高于常规尿细胞学检查;同时FISH在膀胱尿路上皮癌诊断中的敏感性高于ICM,所以FISH技术更有望成为膀胱尿路上皮癌无创性的诊断和检测手段.     

Value of microhematuria, urine cytology, TPA and neopterin in the after care of patients with bladder tumors

Tumor recurrences were observed 70 times in 715 cystoscopies performed in 253 patients. The sensitivity of microhematuria to detect a tumor recurrence was 61%, the specificity 84%. The sensitivity of microhematuria increased to 90% in Tis and T2 tumors. Urine cytology showed a specificity of 100% and a sensitivity of only 43%. The specificity and sensitivity of TPA was only 58% and 41%, respectively, the latter increased to 80% in Tis. 60 patients with proven tumor recurrence showed an increase of neopterin with higher tumor stage. In tumors of stage T2 and Tis serum neopterin was raised in 90% and urine neopterin in 75%. Based on these results cystoscopy, exfoliative urinary cytology and urine analysis are obligatory in the follow-up of patients with superficial bladder cancer. Because of the low specificity (29-41%) TPA and neopterin are not suitable for follow-up.     

The predictive value of multi-targeted fluorescent in-situ hybridization in patients with history of bladder cancer

OBJECTIVES: UroVysion (Abbott Molecular Inc., Des Plaines, IL) is a multi-target fluorescent in-situ hybridization (FISH) assay that detects aneuploidy of chromosomes 3, 7, and 17, and loss of the 9p21 locus in exfoliated cells in urine. In this study, we evaluated if UroVysion can predict tumor recurrence in patients with negative cystoscopy and urinary cytology at the time of (FISH) assay. METHODS: The study population included patients with history of non-muscle invasive bladder cancer treated by transurethral resection. Follow-up included cystoscopy, barbotage, urinary cytology, and UroVysion testing. Patients were followed for at least 6 months after their initial UroVysion testing. RESULTS: A total of 64 patients (37 males) were enrolled into the study. Mean patient age was 62 years (S.D. 13.2 years). Initial highest tumor stage was Ta in 42 patients (65.6%), T1 in 21 patients (33%), and isolated Tis in a single patient. Abnormal UroVysion results were observed in 40 patients (62.5%). After a median follow-up of 13.5 months, 21 patients (33%) developed tumor recurrence (Ta in 13 patients, T1 in 5, and Tis in 3). Recurrent tumors developed in 45% of the patients with abnormal UroVysion test compared with 12.5% of the patients with normal assay (P = 0.01). An abnormal UroVysion result preceded the diagnosis of tumor recurrence in 18/21 cases (86%), including all high-grade recurrences. CONCLUSIONS: This data suggest that UroVysion may be a useful tool for predicting tumor recurrence. Cystoscopy may be spared and surveillance intervals widened in patients with history of low grade tumors and a normal UroVysion test.     

Should Bladder Biopsies be Performed Routinely after Bacillus Calmette-Guérin Treatment for High-Risk Superficial Transitional Cell Cancer of the Bladder?

OBJECTIVE: After endoscopic resection of high-grade superficial urothelial neoplasms (Ta, T1 or Tis), adjuvant bacillus Calmette-Guérin (BCG) therapy is performed routinely to avoid recurrence and/or progression. Vesical biopsies often are performed to assess the efficacy of treatment. The aim of our study was to evaluate the usefulness of these biopsies. MATERIALS AND METHODS: During this retrospective bi-centre study, 130 patients who had undergone vesical high-grade tumour resection were included. There were 40 Ta associated with Tis in three cases, 87 T1 associated with Tis in 13 cases, and three isolated Tis. After BCG treatment, the following parameters were studied: cytoscopic findings, urine cytology and the histologic results of systematised biopsies. RESULTS: Urine cytology was positive (high-grade) for 26 patients and negative (normal or low-grade) for 104 patients. For the 26 patients with positive cytology, vesical flexible cystoscopy findings were considered suspicious in 18 patients and normal in eight patients. As for the 104 patients who presented negative cytology, cystoscopic findings were considered negative in 76 patients and suspicious in 28 patients. In the present study, the sensitivity of cytology and cystoscopy in the detection of recurrence after BCG treatment was 56% and 87.5%, respectively; specificity was 56% and 81.6%, respectively. When the two examinations were combined, sensitivity was 100%, and specificity was 76%. CONCLUSIONS: After BCG therapy, the association of negative flexible cystoscopy findings and normal urine cytology made it possible to avoid routine biopsies. Patients could therefore avoid the morbidity of this procedure.