应用荧光原位杂交技术检测上尿路的尿路上皮癌可显著提高诊断敏感性

论著荧光原位杂交用于上泌尿系统尿路上皮癌的诊断研究丁雪飞高鹰周广臣摘要目的评价F IS H 用于诊断上泌尿系尿路上皮癌(U UT UC)的临床应用价值。

方法应用荧光原位杂交(F IS H )检查和尿细胞学检查58例UUT UC 患者和20例健康人群尿沉渣标本,并比较两者的效果。

F IS H 探针采用随机引物法标记3、7、17号染色体着丝粒及9p21(p16)区带。

分析F IS H 结果中各个探针的出现频率。

结果F IS H 对于UUT 检查敏感性高于尿细胞学检查(759%vs .345%,P =00001),特异性相似(90%vs .95%,P =0548)。

F IS H 和尿细胞学检查阳性预测值956%和952%,阴性预测值分别是563%和333%。

探针中3、7和17号染色体非整倍性扩增率分别为523%、569%和341%,p16杂合性缺失丢失273%。

结论应用3、7、9和p16探针的F IS H 检查用于U UT UC 中敏感性更高而特异性相似,可用于UUT UC 的术前诊断。

关键词泌尿系肿瘤;原位杂交,荧光;细胞诊断学U t ility of f l uor escen ce in situ hybr i d iza ti on i n the d iagnosis of upper ur i na r y tra ct ur othe lia l ca r ci no m aDING Xue fei ,GA O Y ing,Z HOU Guang chen.D e par t m ent o f Urolo gy,The Subei P eo ple sHos p ita l ,Y angzho u 225001,ChinaCorres ponding author:DI NGXue fei ,Em a il :daofeng06@126.co mAbstra ct Ob jec ti ve The purpose of this st udy was to eva l uate t he c linical utility of fl uorescence in sit u hybridiza tio n (F IS H )assay as a non i nvasi ve m e t hod f or diag nosi ng urot he lial carci no ma (UC)i n the uppe r uri nary tract (UUT).M ethod s U rine spec i m ens fro m 58co nsecuti ve patients w ith UUT UC and 20healthy control s were ana l yzed by cytol ogy and F IS H.F or F IS H ana l ysi s ,labe led probes specific f or chro moso m es 3,7,and 17and for the p16(9p21)genes were used to assess chro m oso m al abnor m a li ties ind i cative of m ali gnancy .Sensiti vity and spec ificity of both techniques were deter m i ned and co m pa red .The frequency of chro moso m a l aberra ti ons of m a li gnant cell s fro m UUT was also deter m i ned .R esu lts Overall sens i tivit y of F IS H was s i gnificantl y h i gher than tha t of ur i ne cytol ogy (759%vs 345%,P =00001).Specificities for both F IS H and cytol ogy were si m ilar resu lts (90%vs 95%,P =0548).The positi ve and nega tive pred icti ve values for F IS H were 956%and 563%,whereas for cytol ogy they were 952%and 333%,respecti vely .Of 58pati ents w ith UUT UC ,pol yso m i es of chro m oso m e 3,7and 17were observed i n 523%,569%and 357%,respecti vely ,and loss of the p16gene in 273%.C on clusi ons F IS H assay of chro m oso m es 3,7,9,and 17per f or m ed o n exfoli a ted ce lls fro m voi ded urine speci m ens has grea ter sensiti vity than cytology for detecti ng UUT UC ,and they we re m a i nta i n i ng a si m ilar spec ifi c ity .K ey wor ds U rol ogic neoplas m s ;In situ hybri d izati on ,fl uorescence ;Cytodiagnosis上泌尿系统(the upper ur i nary trac,t U UT)尿路上皮癌(ur othe li a l carc i no mas ,UC)的早期诊断主要依赖于尿细胞学检查联合影像学和/或输尿管镜,但对于早期、直径较小的肿瘤敏感性较低[1]。

山东医药2023 年第 63 卷第 30 期尿液脱落细胞块切片的制备杜天海，吕丽霞，杨洁亮，彭恒，陈敏四川大学华西医院病理科，成都610041摘要：目的　制作尿液脱落细胞块切片，并观察制备效果。

方法　收集36例疑似尿路上皮癌患者的新鲜晨尿样本，离心后取尿液沉渣，裂解沉渣中红细胞后加入固定液固定尿液脱落细胞，将尿液脱落细胞富集后制备尿液脱落细胞团，将尿液脱落细胞团脱水后石蜡包埋，以制备尿液脱落细胞块。

将尿液脱落细胞块切片，苏木素—伊红染色（HE）后显微镜下评估切片细胞数量（一个视野中细胞所占面积>40%为制备成功）。

将制备成功的尿液脱落细胞块切片分别进行免疫组织化学（IHC）和荧光原位杂交（FISH）检测，IHC染色清晰，定位准确，阳性细胞与背景分辨清晰视为检测成功，FISH检测荧光信号对比清晰，背景干净且75%以上细胞核中有杂交信号视为检测成功。

结果　HE染色显微镜下评估后36例份尿液脱落细胞块切片中21例份制备成功、15例份因细胞过少无法满足后续诊断要求。

21例份尿脱落细胞块切片HE染色后显微镜下可见细胞单层平铺，背景干净，核仁清晰，细胞核和胞质对比清晰。

21例份尿脱落细胞块切片IHC染色后荧光显微镜下细胞定位准确，细胞内颗粒清晰，染色强度适宜。

21例份尿脱落细胞块切片FISH染色后荧光显微镜下细胞核形态轮廓清晰，荧光红绿信号对比清晰，背景干净。

结论　成功制备尿液脱落细胞块切片。

尿液脱落细胞块切片后可用于常规HE染色、IHC和FISH检测，显微镜下可见背景干净，细胞染色效果较好，细胞核仁清晰。

关键词：细胞块制备技术；尿液脱落细胞块；尿脱落细胞学检测doi：10.3969/j.issn.1002-266X.2023.30.021中图分类号：R3 文献标志码：A 文章编号：1002-266X（2023）30-0087-03尿路上皮癌是临床上常见的泌尿系统肿瘤之一，近年来发病率呈逐年上升趋势。