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.     

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.     

荧光原位杂交在非肌层浸润行膀胱尿路上皮癌术后随访中的应用

目的:研究多色荧光原位杂交探针(FISH)在非肌层浸润性膀胱尿路上皮癌术后监洲的应用价值,并与尿细胞学检查相比较.方法:应用FISH和尿细胞学检查分析215例非肌层浸润性膀胱尿路上皮癌术后随访尿沉渣标本.FISH探针是用随机引物法标记3、7、17号染色体着丝粒及9p21区带.比较分析两种检查结果.结果:215例中有58例患者病理证实膀胱尿路上皮癌复发.FISH对膀胱尿路上皮癌诊断高于尿脱落细胞学检查(71.4%和26.2 %),并有统计学差异(P<0.05),特异性分别为90%和95%,无统计学差异;阳性预测值分别是95.5%和95.7%,阴性预测值分别为52.9%和34.5%.结论:FISH检查可以作为非肌层浸润性尿路上皮癌术后随访的手段,并且敏感性高于尿细胞学检查.     

血尿的病因分析及荧光原位杂交技术用于诊断尿路上皮癌的价值

目的总结并分析初发血尿的病因,同时比较荧光原位杂交技术(FISH)与尿脱落细胞学检查技术在诊断尿路上皮癌(UC)中的敏感性和特异性,从而进一步评估FISH用于筛查尿路上皮癌的临床价值。方法回顾性分析500例入院诊断为"血尿待查"患者的最终出院诊断结果,同时记录其中每例患者的检查诊治经过。计算比较FISH与尿脱落细胞学检查的敏感性与特异性。结果 500例初次诊断血尿待查患者中最终诊断为尿路上皮癌130例、泌尿系结石30例、泌尿系感染135例、前列腺癌12例、前列腺增生(BPH)相关出血66例、乳糜血尿23例、特发性血尿60例、其他44例。有218例诊疗过程中同时采用了FISH检测和尿脱落细胞学检查;FISH敏感性明显优于脱落细胞学(75.4%vs.33.9%,P0.001),两者特异性无明显差异(96%vs.99%,P=0.837)。对于上尿路尿路上皮癌(UTUC)的诊断,FISH和尿脱落细胞学检查敏感性分别为80.4%和34.8%(P0.001)。结论血尿待查病因复杂多样,需要临床医生扎实的基础和临床理论知识及正确的临床思维能力。FISH对于尿路上皮癌尤其是上尿路尿路上皮癌的诊断具有重要价值。     

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.     

荧光原位杂交技术在膀胱尿路上皮癌中的临床应用研究

目的:本研究旨在利用荧光原位杂交技术(FISH)辅助诊断膀胱尿路上皮癌,以探讨其作为一种替代尿脱落细胞学的无创性检查在膀胱尿路上皮癌诊断中的应用价值。方法:收集30例健康志愿者的新鲜尿液,应用3、7、17号染色体及9号染色体p16位点探针标记,进行FISH检测,建立正常人群阈值;再收集95例因血尿或影像学检查拟诊为膀胱肿瘤的患者晨尿标本,分别作常规尿脱落细胞学检查和FISH检测,其中79例膀胱镜下活检或术后标本病理确诊为膀胱尿路上皮癌。FISH检测统计目标染色体畸变情况,并进一步分析与病理分级的关系。结果:FISH检测设定的正常阈值为3号染色体:3%;7号染色体:4%;17号染色体:3%;9号染色体p16位点:10%。FISH检测技术的敏感性为74.68%(59/79),高于尿脱落细胞学检查的35.44%(28/79),差异具有统计学意义(P>0.001),而FISH检测的特异性为93.75%(15/16),尿脱落细胞学检查特异性为87.50%(14/16),两者差异无统计学意义(P>0.05)。不同病理分级下FISH检测的敏感性呈现为高级别高阳性率:低度恶性倾向的尿路上皮肿瘤为58.33%(7/12),尿路上皮癌Ⅰ级为54.17%(13/24),尿路上皮癌Ⅰ～Ⅱ级为86.36%(19/22),尿路上皮癌Ⅱ级和Ⅱ～Ⅲ级为94.44%(17/18),尿路上皮癌Ⅲ级为100%(3/3),敏感性随着病理分级的提高而呈现增高趋势。结论:FISH检测技术具备无创、高敏感性的特点,优于传统的尿脱落细胞学检查。不同分级的膀胱尿路上皮肿瘤其FISH检测的敏感性不同,表现为高级别高敏感性。FISH检测上述位点在低级别膀胱尿路上皮肿瘤中敏感性较高,大大提高了低级别尿路上皮肿瘤的检出率,有望成为一种常规的膀胱肿瘤复查指标。     

荧光原位杂交技术在膀胱肿瘤诊断中的应用

目的：评价荧光原位杂交技术（FISH）在膀胱肿瘤诊断中的应用价值。方法：20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,建立阈值。20例血尿患者留取晨尿,分别进行尿脱落细胞学检查和FISH检查。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。结果：20例血尿患者除1例为泌尿系结石、1例为BPH外,病理检查证实的18例膀胱肿瘤患者中,FISH检出16例,尿脱落细胞学检出4例。结论：FISH无创快速,检出率高,在膀胱肿瘤早期诊断中优于尿脱落细胞学检查方法。     

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.     

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。     

Role of fluorescence in situ hybridization in bladder cancer surveillance of patients with negative cytology

ObjectivesThe clinical utility of urine markers in urothelial cancer (UC) surveillance is not established. We previously evaluated the use of fluorescence in situ hybridization (FISH) in managing patients with atypical cytology at risk for UC. This study evaluates its role in patients with negative cytology with a history of UC.Materials and methodsBetween June 2007 and January 2009, every patient with a history of UC who underwent cystoscopy and cytology with UroVysion test were identified. A comprehensive chart review was performed on each patient with negative cytology.ResultsThe population comprised 142 patients undergoing cancer surveillance; 111 patients with negative cystoscopy, 19 with equivocal cystoscopy, and 12 with positive cystoscopy. In patients with negative cystoscopy, there was cancer in only 1 of 111 patients. UroVysion could detect the only patient with UC with sensitivity of 100% and had a negative predictive value (NPV) of 100%. In patients with equivocal cystoscopy, it detected 2 tumors that would be missed by cytology. There were 4 false negative results (sensitivity 33.3% and NPV 66.7%). In patients with obvious lesion on cystoscopy, there were 9 false negative results (sensitivity 10% and NPV 18.2%).ConclusionsFew patients with negative cystoscopy and negative cytology have cancer. Patients with equivocal and positive cystoscopy and negative cytology frequently have cancer and the UroVysion FISH assay was not helpful in these cases. The cost-effectiveness of the FISH assay needs to be assessed prior to widespread use in patients with negative cytology.     

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:评估荧光原位杂交技术(FISH)在膀胱尿路上皮癌诊断中的应用价值。方法:收集60例疑似膀胱癌的血尿患者的尿液标本,分别作尿细胞学检测和荧光原位杂交分析。20例正常人尿液标本,用于建立FISH阀值,作为阳性判断的标准。结果:细胞学和FISH的总敏感性分别为42.0%、82.2%,特异性分别为:93.3%、86.7%。细胞学和FISH在低级别及非肌层浸润性肿瘤等敏感性的均差异有统计学意义(P<0.05)。结论:FISH技术能明显提高膀胱尿路上皮癌的检出率,尤其是早期和低级别病变,可以成为筛查膀胱尿路上皮癌的一种新的无创性检查方法。     

A comparison of cytology and fluorescence in situ hybridization for the detection of urothelial carcinoma

PURPOSE: We determine the relative sensitivities of cytology and fluorescence in situ hybridization (FISH) for the detection of urothelial carcinoma. MATERIALS AND METHODS: A mixture of fluorescent labeled probes to the centromeres of chromosomes 3, 7 and 17, and band 9p21 (P16/CDKN2A gene) was used to assess urinary cells for chromosomal abnormalities indicative of malignancy. A total of 280 urine specimens from 265 patients, including 150 with a history of urothelial carcinoma and 115 without a history of urothelial carcinoma, were analyzed. FISH analysis was performed without prior knowledge of clinical findings, that is biopsy, cystoscopy and cytology results. A positive result was defined as 5 or more urinary cells with gains of 2 or more chromosomes. RESULTS: A total of 75 biopsies showed urothelial carcinoma at FISH analysis among the 265 patients. The sensitivity of urine cytology for pTa (36 cases), pTis (18) and pT1-pT4 (15) tumors was 47%, 78% and 60%, respectively, for an overall sensitivity of 58%. The sensitivity of FISH for pTa (37 cases), pTis (17) and pT1-pT4 (19) tumors was 65%, 100% and 95%, respectively, for an overall sensitivity of 81%. FISH was significantly more sensitive than cytology for pTis (p = 0.046), pT1-pT4 (p = 0.025), grade 3 (p = 0.003) and all tumors (p = 0.001). The specificity of cytology and FISH among patients without cystoscopic evidence of urothelial carcinoma and no history of urothelial carcinoma was 98% and 96%, respectively (p = 0.564). CONCLUSIONS: The sensitivity of FISH for the detection of urothelial carcinoma is superior to that of cytology, and the specificity of FISH and cytology for urothelial carcinoma are not significantly different. Further prospective studies are required but FISH has the potential to improve significantly the management of urothelial carcinoma.     

5-氨基乙酰丙酸诱导荧光膀胱镜在膀胱肿瘤诊断中的应用（附31例报告）

目的：研究5-氨基乙酰丙酸（5-ALA）诱导荧光光动力学对膀胱肿瘤的早期诊断价值。方法：对血尿患者行5-ALA诱导荧光膀胱镜检查及活组织检查,以5-ALA膀胱灌注,2h后采用D-light光源系统进行膀胱镜检,对荧光阳性区域及白光下肉眼可见异常但荧光阴性区域进行活检,活检后行经尿道膀胱肿瘤电切术。结果：31例患者中有4例荧光阴性且普通光肉眼观阴性者未活检。余27例患者共取活检96处,其中荧光阳性区域取活检89处（包括普通光肉眼观阴性区域35处）,切缘取活检7处。病理检查结果显示：尿路上皮癌65处,阳性率为73．03％（65／89）,非肿瘤性病变24例,假阳性率为27％（24／89）,切缘活检7处为阴性。荧光下阳性而白光下阴性的肿瘤位点11处,切缘活检7处均为阴性。结论：5-ALA诱导荧光光动力学诊断对膀胱肿瘤有较高价值,能发现早期肿瘤,同时进行电切将更彻底。     

Novel NMP22 quick qualitative analysis (BladderChek NMP22) for the diagnosis of urothelial tumor

PURPOSE: The BladderChek NMP22 test was studied to determine whether it could be a useful diagnostic tool for urothelial tumor. MATERIALS AND METHODS: During the period from April to May 2004, a total of 50 patients who visited Nagano Municipal Hospital were subjected to analysis with the BladderChek NMP22 test. They consisted of 25 patients with microscopic or gross hematuria and 25 patients with urothelial tumor who were being followed after surgery. The clinical diagnosis was made by cystoscopy, abdominal CT scanning, ultrasonography or intravenous urography. BladderChek NMP22 was used in all of the patients and the test was judged to be positive when a line appeared on the glass at 30 minutes after a drop of urine was added to the sample well. Cytological examination of the urine was also done. It was defined as negative if the result was I, II or III, while it was positive if the result was IV or V. RESULTS: Seven patients who were not eligible were excluded and the remaining 43 patients were studied. Eleven of the 43 patients were diagnosed as having urothelial tumor. The sensitivity, specificity, and accuracy of BladderChek NMP22 for making a correct diagnosis was 63.6%, 62.5%, and 62.7%, respectively (p = 0.1703). As for urine cytology, the sensitivity, specificity, and accuracy was 36.3%, 100%, and 83.7%, respectively (p = 0.0027). BladderChek NMP22 was positive in all the patients whose urine cytology was positive and in three patients with low-grade, low-stage, and small tumors, although urine cytology was negative. On the other hand, false-positive BladderChek NMP22 results were obtained in patients undergoing follow-up after removal of urothelial carcinoma and in patients with urinary stones. The sensitivity of BladderChek NMP22 was superior to that of urine cytology, but its specificity was inferior. CONCLUSION: The combined use of BladderChek NMP22 and cystoscopy may be useful for the diagnosis and follow-up of urothelial tumor. On the other hand, the results obtained with BladderChek NMP22 need to be interpreted carefully because the test can yield a false-positive outcome in the presence of marked hematuria, pyuria or cytolysis of normal urotherium. When the result of BladderChek NMP22 in the monitoring of recurrent urotherial tumor following surgery was positive in spite of negative findings of cystoscopy, it is necessary to consider the presence of a tumor in the upper urinary tract or a high tendency toward tumor recurrence. In such a case, careful followup may be necessary.     

荧光原位杂交技术检测膀胱尿路上皮癌尿液的临床应用研究

目的：评价荧光原位杂交技术（fluorescence in situ hybridization,FISH）检测膀胱尿路上皮癌患者尿液的应用价值。方法：收集我院2007年10月-2009年4月期间77例膀胱尿路上皮癌患者、43例非尿路上皮癌的血尿患者（通过膀胱镜检查排除尿路上皮癌）和泌尿系良性疾病患者的晨尿,同时行FISH检测和尿脱落细胞学分析,再结合病理结果将两种方法进行比较。FISH检测使用荧光标记DNA探针混合物与细胞核上3、7、17号染色体着丝粒和9p16位点进行杂交。结果：FISH总的敏感度和特异度分别为89．6％和95．3％,G1-3各级的敏感度分别为76．1％、90．9％、100％,Ta、Tis、T1、T2-4各期的敏感度分别为55．6％、100％、88．9％、97．4％。尿脱落细胞学分析总的敏感度和特异度分别为37．7％和93．0％,G1-3各级的敏感度分别为0％、33．3％、78．3％,Ta、Tis、T1、T2-4各期的敏感度分别为11．1％、100％、14．8％、56．4％。结论：FISH比尿脱落细胞学提高了膀胱癌患者检测的敏感度,而特异度两者相近。FISH使低级别浅表型膀胱癌的准确率明显提高,几乎能检测出所有高级别的浸润型膀胱癌。相对于尿脱落细胞学,FISH检测更佳。     

荧光原位杂交技术尿脱落细胞检测在上尿路尿路上皮细胞癌诊断中的应用价值

目的 探讨荧光原位杂交技术( FISH)尿脱落细胞检测在上尿路尿路上皮细胞癌诊断中的应用价值.方法 病理诊断为上尿路尿路上皮细胞癌患者30例,留取新鲜晨尿,行FISH和尿脱落细胞学检查,并行多层螺旋CT和彩色多普勒超声检查;留取20例肾癌、6例输尿管结石和4例输尿管狭窄患者的尿液作对照,统计学分析FISH和尿脱落细胞学检查诊断的特征值.结果 FISH、尿脱落细胞学、CT和超声检查诊断上尿路尿路上皮细胞癌的敏感性分别为87％( 26/30)、37％(11/30)、90％(27/30)和43％ (13/30),FISH和CT诊断敏感性差异无统计学意义(P＞0.05),但明显高于超声和尿脱落细胞学检查(P＜0.05);FISH和尿脱落细胞学检查诊断的特异性分别为97％(29/30)和93％ (28/30),二者比较差异无统计学意义(P＞0.05).FISH和尿脱落细胞学检查阳性预测值分别为96％(26/27)和85％ (11/13),阴性预测值分别为88％(29/33)和60％( 28/47).结论 FISH诊断上尿路尿路上皮细胞癌具有较高的敏感性、特异性、阳性预测值和阴性预测值,可作为上尿路尿路上皮细胞癌诊断和术后随访检查的有效方法.     

MRI联合荧光原位杂交技术在尿路上皮癌术后复发监测中的应用分析

目的 评价MRI联合荧光原位杂交技术(fluorescence in situ hybridzation,FISH)和FISH、膀胱镜检查在尿路上皮癌术后复发监测中的应用,为完善尿路上皮癌术后复发监测提供理论依据.方法 选取2013年1月至2015年1月60例临床诊断的高危膀胱尿路上皮癌患者,术后第3、6、9、12和18个月随访均行FISH、FISH联合MRI检查及膀胱镜加活组织检查.结果 60例入围患者的监测,单纯采用FISH检查,阳性33例,疑似肿瘤复发占55.0%;MRI加FISH检查,阳性累计36例,疑似肿瘤复发占60.0%.所有疑似病例均采用膀胱镜加活组织检查,最终确认肿瘤复发患者35例,占全部患者的58.3%,其中单纯采用FISH检查发现肿瘤复发27例,占肿瘤复发患者的77.1%;而采用MRI联合FISH检查发现肿瘤复发累计达32例,占肿瘤复发患者的91.4%.结论 MRI联合FISH对膀胱肿瘤检测的阳性率高于单用FISH检测,MRI联合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.