宫颈脱落细胞不同制片方法对FISH检测TERC基因实验结果的影响  

Compariation of Sampling and Processing Procedures of Cytological Specimens of the Cervical Exfoliated Cell for FISH Study

作　　者：汤冬玲[1] 李艳[1] 段峰[1] 明蕾[1] 刘玉凌[1] 

机构地区：[1]武汉大学人民医院,武汉430060

出　　处：《现代检验医学杂志》2009年第6期56-58,共3页Journal of Modern Laboratory Medicine

基　　金：本课题受卫生部科研基金项目（WKJ2007-3-001）资助.

摘　　要：目的探讨利用双色间期荧光原位杂交技术（fluorescence in situ hybridization，FISH）检测人宫颈脱落细胞端粒酶（TERC）基因不同制片的最佳方法。方法采集门诊就诊的114例患者的宫颈脱落细胞，分另q采用新鲜细胞直接涂片（45例）、液基薄层细胞学（TCT）剩余液体直接制片（37例）和TCT低渗制片（32例）三种取材制片方法，应用双色间期FISH法检测宫颈脱落细胞的TERC基因表达，对比取材刺片的最佳方法。结果在直接涂片法、TCT制片法和TCT低渗法三种制片过程中，TERC基因的杂交成功率分别是55．1％，34．7％，81．3％，其中TCT低渗组杂交成功率明显高于其他两组，差异有统计学显著性意义（P〈0．05）。直接涂片法、TCT制片法、TCT低渗制片法的平均裸核细胞比例分别为24％，15％，62％，TCT低渗法高于其他两种方法，与其他两组比较差异有统计学显著性意义（P〈0．05）。三种制片方法的玻片背景清晰度比例分别为33．3％，35．2％和68．8％。结论应用双色间期FISH技术检测宫颈脱落细胞TERC基因，以TCT低渗制片法简便快速，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 RNA eornponent gene （TERC） on cytological samples of the cervix. Methods Cervical specimens from 114 outpatients were obtained and three groups were divided based on the different sampling and processing procedures ,including cervical Pap smears （45 cases）,thin-layer slides （37 cases） and hypotonic thin-layer slides （32 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, regular thin-layer slides and hypotonic thin-layer slides were 55.1% ,34.7% and 81.3% ,respectively. The hybridization rate in the hypotonic thin-layer slides was significantly higher than that in the thin-layer slides （P〈0. 05）. The naked nucleolus rates among the three groups were 24 %, 15 % and 62%, respectively. The slide background clearness rates of the above three groups of slides were 33.3% ,35.2% and 68. 8% ,respectively. Among these methods,the slides clearness by Pap smears was significant different with that by TCT （P〈 0. 05）. Conclusion The hypotonie thin-layer approach is simple,quick and also generate the highest hybridization successful rates when detecting the TERC gene by dual-color interphase FISH technique. It is suitable for cytological and molecu- lar cytogenetic analysis of the cervical cells.

关 键 词：荧光原位杂交 宫颈脱落细胞 TERC基因 

分 类 号：R392.12[医药卫生—免疫学]