title: Improvement of specific detection of circulating tumor cells using combined CD45 staining and fluorescence in situ hybridization.
abstract: BACKGROUND
  Conventional cytokeratin (CK)-based methods to detect circulating tumor cells (CTCs) often present suboptimal sensitivities for decreased or non-expression of cytokeratin in some CTCs. We clinically investigated a new method that combines immunocytochemistry staining (ICC) of CD45 and fluorescence in situ hybridization (FISH).
  
  
  METHODS
  Circulating epithelial cells from 141 subjects were enriched using an EpCAM-independent strategy and then identified by either a combination of FISH with chromosome 8 centromere probe (CEP8) and ICC staining of CD45 (CD45-FISH) or ICC staining of CK.
  
  
  RESULTS
  For detecting CTCs enriched from lung cancers, CD45-FISH had larger areas under ROC curves of 0.963 (P=0.000) compared to ICC (0.653; P=0.031) using cut-off values of 2 and 1 cell/3.75ml blood with sensitivities of 83.3% and 43.3%, specificities of 98.6% and 89.5%, respectively. Moreover, CD45-FISH showed 76.2% sensitivity in detecting CTCs in ovarian cancers (P<0.001). Four of six ovarian cancers showed dramatical decrease in both CTCs and serum CA125 on the 7th day after surgery.
  
  
  CONCLUSIONS
  CD45-FISH method had improved sensitivity and specificity in detecting CTCs of lung and ovarian cancers compared to ICC-CK. This combined detection strategy may be useful in detecting or monitoring CTCs after ovarian cancer surgery.
authors.affiliation: Department of Clinical Laboratory, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, China; Department of Obstetrics and Gynecology, The First Affiliated Hospital of Harbin University, Harbin, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Ning
authors.affiliation: Department of Clinical Laboratory, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Zhan
authors.affiliation: Department of Clinical Laboratory, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Yujuan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Clinical Laboratory, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Chen
authors.affiliation: Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Fengzhi
authors.initials: F
authors.lastname: Feng
authors.affiliation: Department of Clinical Laboratory, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Department of Clinical Laboratory, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Zhongjuan
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Department of Clinical Laboratory, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Danfei
authors.initials: D
authors.lastname: Xu
authors.affiliation: Department of Clinical Laboratory, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Wang
authors.affiliation: Department of Clinical Laboratory, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Department of Clinical Laboratory, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Xing
authors.affiliation: Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Guan
authors.affiliation: Department of Clinical Laboratory, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, China. Electronic address: wendycuiwei@hotmail.com.
authors.forename: Wei
authors.initials: W
authors.lastname: Cui
mesh_terms: D002469:Cell Separation / Q000379:methods*; D005260:Female; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D017493:Leukocyte Common Antigens / Q000378:metabolism*; D008175:Lung Neoplasms / Q000473:pathology; D009360:Neoplastic Cells, Circulating / Q000378:metabolism / Q000473:pathology*; D010051:Ovarian Neoplasms / Q000473:pathology; D015203:Reproducibility of Results
chemical_list: D017493:Leukocyte Common Antigens
keywords: CD45-FISH; CTCs; ICC; Lung cancer; Ovarian tumor