title: Real-time quantitative RT-PCR detection of circulating tumor cells from breast cancer patients.
abstract: Circulating tumor cells (CTCs) were recognized as novel tumor biomarker for prognostic and predictive purposes in various cancers. Various detection technologies and devices have been developed to enumerate and characterize CTCs. Most of those approaches are based on the positive enrichment strategy and immunocytological techniques. However, the sensitivity of these approaches proved to be limited in metastatic tumors and the detection of early tumor cell dissemination was problematic. In the present study, we developed a novel CTC detection method by real-time RT-PCR technique in combination of negative enrichment strategy. The developed enrichment approach could recover more than 75% of spiked breast cancer cells from peripheral blood. The detection limit of duplex real-time RT-PCR assay using KRT19 and ERBB2 as targeted genes was consistently one breast tumor cell. Moreover, CTC detection by duplex real-time RT-PCR assay had higher detection sensitivity than that by immunostaining, especially in early breast cancer. In summary, the results of the present study indicated the potential clinical utilities of CTCs identification on breast cancer by duplex real-time RT-PCR in combination with negative enrichment.
authors.affiliation: School of Pharmacy, Zhengzhou University, Zhengzhou, Henan 450001, P.R. China.
authors.forename: Maowen
authors.initials: M
authors.lastname: Guo
authors.affiliation: School of Pharmacy, Zhengzhou University, Zhengzhou, Henan 450001, P.R. China.
authors.forename: Xiaotian
authors.initials: X
authors.lastname: Li
authors.affiliation: Affiliated Hospital of Academy of Military Medical Sciences, Beijing 100071, P.R. China.
authors.forename: Shaohua
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Affiliated Hospital of Academy of Military Medical Sciences, Beijing 100071, P.R. China.
authors.forename: Hua
authors.initials: H
authors.lastname: Song
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, P.R. China.
authors.forename: Wenhui
authors.initials: W
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, P.R. China.
authors.forename: Xueyi
authors.initials: X
authors.lastname: Shang
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, P.R. China.
authors.forename: Yuling
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, P.R. China.
authors.forename: Hua
authors.initials: H
authors.lastname: Jiang
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, P.R. China.
authors.forename: Qingyu
authors.initials: Q
authors.lastname: Lv
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, P.R. China.
authors.forename: Yongqiang
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, P.R. China.
authors.forename: Huaijie
authors.initials: H
authors.lastname: Hao
mesh_terms: D014408:Biomarkers, Tumor / Q000097:blood / Q000235:genetics*; D001943:Breast Neoplasms / Q000235:genetics / Q000473:pathology*; D002469:Cell Separation / Q000379:methods; D004273:DNA, Neoplasm / Q000032:analysis*; D005260:Female; D018922:HL-60 Cells; D006801:Humans; D019169:Jurkat Cells; D061986:MCF-7 Cells; D009360:Neoplastic Cells, Circulating / Q000378:metabolism / Q000473:pathology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D014407:Tumor Cells, Cultured
chemical_list: D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm