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。     

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尿路上皮癌预警诊断中的应用

目的：探讨使用荧光原位杂交技术对尿路上皮癌进行预警诊断的可行性和有效性。方法：采用3、7、17号染色体着丝粒探针和9p21区带探针对30例影像学检查、膀胱镜检查、尿脱落细胞学检查均为阴性的高度可疑的尿路上皮癌的血尿患者尿液脱落细胞核行荧光原位杂交检测。结果：30例血尿患者中,11例FISH结果阳性。随访3～13个月。有5例确诊为膀胱尿路上皮癌,2例确诊为肾盂癌。19例FISH检测阴性的患者无一例患病。结论：尿脱落细胞荧光原位杂交技术对膀胱镜和尿脱落细胞学检查阴性的早期尿路上皮癌患者有预警诊断作用．具有重要的临床应用价值。     

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.     

Study on the diagnosis of urothelial cancer using multi-colour fluorescence in situ hybridization (FISH)--comparative analysis between FISH and cytology

To determine the diagnostic merit or demerit of genetic procedures using fluorescence in situ hybridization (FISH) for detecting both new and recurrent urothelial cancer, we analyzed the specimens from 81 out-patients with asymptomatic haematuria, aged over 40, in comparison with urine cytology. Of 10 with atypical cytology, 6 showed positive for FISH, and of these, 4 manifested urothelial cancer. FISH showed higher sensitivity in low/intermediate grade cases compared with cytology (FISH; 66.7% vs cytology; 11.1%). Of 15 primary bladder cancer, 4 showed recurrence, and all of these cases showed a positive FISH reaction, but only 1 in cytology. The sensitivity, specificity and accuracy of FISH tests were 81.2, 72.3 and 74.1%, respectively, and these of cytology were 37.5, 98.5 and 86.4% respectively. The FISH test was superior to cytology for sensitivity, but specificity and accuracy were inferior. The FISH tests could be a potent procedure for detecting urothelial cancer in cases of low/intermediate grade, atypical cytology and surveillance setting.     

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.     

Clinical application of NMP22 and urinary cytology in patients with hematuria or a history of urothelial carcinoma

For evaluation of the clinical application of immunoassay for nuclear matrix protein 22 (NMP22 immunoassay) and urinary cytology for early diagnosis and detection of bladder cancer in patients with hematuria and/or a previous history of bladder cancer, 209 urine samples obtained from 137 patients presenting episodes of hematuria or a history of bladder cancer were assayed for NMP22 levels and/or prepared for cytology examination. Biopsy was performed when any visible tumor was identified during cystoscopy examination. The median NMP22 concentrations measured in samples taken from patients with active bladder cancer, from patients with a history of bladder cancer but no active disease, from patients with hematuria, and from healthy volunteers were 18.95, 5.45, 6.39, and 3.75 U/ml, respectively. The urinary NMP22 level recorded for patients with urothelial carcinoma was significantly higher than that noted for individuals without active disease. The sensitivity of the NMP22 assay and of urinary cytology in diagnosing bladder cancer was 69% and 67%, respectively. In contrast, the specificity of these two diagnostic modalities reached 72% and 93%, respectively. The NMP22 assay is slightly more sensitive but less specific than urinary cytology in detecting bladder cancer. This study indicates that determination of urinary NMP22 levels is a useful and noninvasive tool for the detection of bladder cancer because of its high sensitivity. The urinary NMP22 assay may be used as a first-line routine screening method; however, it cannot replace the use of urinary cytology because of its lower specificity.     

Immunocytology in the assessment of patients with asymptomatic hematuria

Painless hematuria has remained a diagnostic challenge in daily urological practice. Key problem in the assessment of these patients is the discrimination between malignant and non-malignant conditions. In this prospective study the role of immunocytology in the evaluation of patients with hematuria was investigated. Ucyt is a commercially available immunocytological assay based upon microscopical detection of tumor-associated antigens on the membrane of urothelial cells by immunofluorescence. Between October 2000 and July 2007, 301 consecutive patients with a first episode of painless hematuria without prior transitional cell carcinoma were included. Urine samples were obtained from all patients and examined cytologically and immunocytologically. Clinical assessment by physical examination, laboratory tests, endoscopy and imaging in 228 cases with microhematuria and 66 cases with gross hematuria yielded bladder cancer in 10 (4.6%) and 17 (27%) patients, respectively. Clinical workup demonstrated that composition of both groups was entirely different. Sensitivity of cystoscopy and immunocytology was similar in both groups. Furthermore, a negative finding in cystoscopy and immunocytology virtually excluded the presence of urothelial cancer. However, while predictive values of immunocytology were clearly superior to cytology in gross hematuria, cytology performed better in the microhematuria cohort. Combination of cystoscopy and immunocytology yield 100% sensitivity in the assessment of patients with painless hematuria. Based upon performance characteristics the authors recommend to replace urine cytology by a more sensitive marker like immunocytology in gross hematuria. In patients with microhematuria immunocytology could be used to select for patients at risk for urothelial cancer and thus spare negative patients from further examinations.     

Utility of fluorescence in situ hybridization as a non-invasive technique in the diagnosis of upper urinary tract urothelial carcinoma

OBJECTIVES: To assess the clinical utility of a fluorescence in situ hybridization (FISH) assay as a non-invasive method for diagnosing and monitoring urothelial carcinoma (UC) in the upper urinary tract (UUT). METHODS: Urine specimens from 30 consecutive patients with UUT UC and 19 healthy controls were analyzed by means of cytology and FISH. For FISH analysis, labelled probes to chromosomes 3, 7, 9, and 17 were used to assess chromosomal abnormalities indicative of malignancy. Sensitivity and specificity of both techniques were determined and compared. The frequency of chromosomal aberrations of malignant cells from UUT was also determined. RESULTS: Overall sensitivity for FISH was significantly higher than the corresponding value for urine cytology (76.7% vs. 36%, respectively, p=0.0056). Specificities for FISH and cytology were 94.7% and 100%, respectively (p=ns). The positive and negative predictive values for FISH were 95.8% and 72%, whereas for cytology they were 100% and 54%, respectively. Of the genetically altered nuclei counted, 67%, 54%, and 43% presented polysomy in chromosomes 3, 7, and 17, respectively, and 21% presented a homozygous deletion of chromosome 9. CONCLUSIONS: FISH assay of chromosomes 3, 7, 9, and 17 performed on exfoliated cells from voided urine specimens has greater sensitivity than cytology for detecting UUT UC whilst maintaining a similar specificity. The non-invasive nature of this method and its higher sensitivity could contribute to improving the current diagnosis of UUT UC.     

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.     

NMP22 is a sensitive, cost-effective test in patients at risk for bladder cancer

PURPOSE: The early diagnosis of bladder cancer is central to its effective treatment. This study was designed to determine the clinical use of NMP22 as a urinary marker for the early detection of transitional cell carcinoma of the bladder in patients with hematuria or other indications at risk for malignancy. The sensitivity and specificity of the NMP22 test were compared with urinary cytology, and the results of both tests were then compared to cystoscopic findings. We also determined if NMP22 provided a cost advantage over our current modalities in our patient population. MATERIALS AND METHODS: Each patient submitted a single voided urine which was divided in 2 parts, of which 1 was stabilized with the NMP22 urine collection kit and 1 was preserved in the appropriate cytology medium for cytopathological testing. All patients provided the urine samples before cystoscopic examination. Of the 330 patients 114 (34.5%) presented with microscopic hematuria and 66 (20.4%) with gross hematuria. Other indications for cystoscopy included atypical cytology or unexplained voiding symptoms refractory to medical therapy. RESULTS: There were 18 patients with biopsy confirmed bladder cancer and 312 with benign conditions of the bladder. Median NMP22 value for the malignant bladder tumors was 31.6 units per ml. (95% confidence interval 13.4 to 90.9) and 4.3 units per ml. (3.8 to 4.8) for benign conditions of the bladder. The urinary NMP22 values in the bladder cancer group were significantly higher than those in the benign condition group (p <0.001). The sensitivity of NMP22 was 100% with a specificity of 85% at a reference value of 10.0 units per ml., while cytology had a sensitivity of only 33% and specificity of 100%. Given a negative predictive value of 100% for NMP22, a cost savings of $28,302 to $111,072 (depending on the type of insurance carrier) would have been achieved if it was used alone as the indication for cystoscopy. CONCLUSIONS: This study indicates that urinary NMP22 is a simple, noninvasive, cost-effective marker for the detection of bladder cancer.