应用双色间期FISH技术中宫颈脱落细胞取材及制片方法的探讨  

Evaluation of sampling and processing procedures of cytological specimens of the cervix for FISH analysis

作　　者：李静然[1] 魏丽惠[1] 刘宁[1] 赵丽君[1] 韦平[1] 赵超[1] 崔淑慧[1] 屠錚[1] 赵昀[1] Zhong Chen 

机构地区：[1]北京大学人民医院妇科 [2]Division of medical Genetics,The University of Utah USA

出　　处：《中国妇产科临床杂志》2007年第6期435-438,I0001,共5页Chinese Journal of Clinical Obstetrics and Gynecology

基　　金：卫生部科研基金项目(WKJ2007-3-001);北京大学医学部"985"二期学科建设项目(985-2-015-24)资助

摘　　要：目的在双色间期荧光原位杂交技术(FISH)中探讨采集宫颈脱落细胞样本及制片的最佳方法。方法采集在北京大学人民医院门诊就诊174例患者的宫颈脱落细胞,应用新鲜细胞直接涂片(60例)、生理盐水制片(31例),TCT剩余液体直接制片(37例)、TCT盐水制片(30例)和TCT低渗制片(16例)五种取材制片方法;并应用双色间期FISH法,检测宫颈脱落细胞的TERC基因表达率,对比取材制片的最佳方法。结果在直接涂片法、盐水制片法、TCT制片法、TCT盐水制片法和TCT低渗法五种制片过程中,TERC基因的杂交成功率分别是55.0%、61.3%、37.8%、53.3%和81.3%,其中TCT低渗法杂交成功率与TCT制片组比较,有显著差异(P<0.05),与其他方法比较,无明显差异(P>0.05);5种制片方法的玻片背景清晰比例分别为38.3%、54.8%、35.1%、60%和75%,各组之间有明显差异(P<0.05);5组的无/少信号比例分别为18.3%、16.1%、37.8%、23.3%和18.8%,其中新鲜制片的直接涂片法和盐水制片法与TCT直接制片法比较,有明显差异(P<0.05),与其他方法比较无差异(P>0.05)。结论在应用双色间期FISH技术检测宫颈脱落细胞TERC基因时,以TCT低渗制片和新鲜细胞盐水制片操作方法简单快速,杂交成功率最高。Objective To evaluate the optimal sampling and processing procedures for dual - color interphase fluorescence in situ hybridization （FISH） studies of the human telomerase gene （TERC） on cytological samples of the cervix. Methods Cervical specimens from 174 outpatients were obtained from the People＇ s hospital of Peking University. Five groups were divided based on the different sampling and processing procedures, including cervical Pap smears （60 cases）, brine Pap smears （31 cases）, thin - layer slides （37 cases）, brine thin - layer slides （30 cases）, and hypotonic thin - layer slides （16 cases）. The fluorescent signals in cytological samples of the cervix which detected by using interphase FISH with the dual - color TERC probe were compared among the groups to determine the optimal procedure. Results The successful hybridization rates in Pap smear slides, brine Pap smear slides, regular thin - layer slides, brine thin - layer slides and hypotonic thin - layer slides were 55%, 61.3%, 37.8%, 53.3% and 81.3%, respectively. The hybridization successful rate in the hypotonic thin- layer slides was 81.2%, significantly higher than that in the thin - layer slides （P〈0. 05）, but no significant difference was found when compared with other groups （P〉0.05）. The slide background -clearness rates of the above five groups of slides were 75%, 54. 8%, 38.3%, 60%, and 35. 1%, respectively （significantly difference was found, P〈0. 05）. The percentage of no/less signal were 18. 3%, 16. 1%, 37.8%, 23. 3% and 18. 8%, respectively. Among these methods, the slides clearness by Pap smears and brine Pap smear were significant differ with that by TCT （P〈0.05）, but no significant difference was found when compared with other methods （P〉 0. 05）. Conclusions When detect the TERC gene by dual - color interphase FISH technique, the hypotonic thin - layer and brine Pap smear approaches are simple and quick, and also generate the highest hybridization successful rates. It is suitable

关 键 词：荧光原位杂交 宫颈脱落细胞 实体肿瘤 TERC基因 

分 类 号：R446.8[医药卫生—诊断学]