Establishment and Application of Immunomagnetic Bead Negative Enrichment and Immunofluorescence Antibody Technique for Detection of CTCs in Peripheral Blood of Patients with Ovarian Cancer / 现代检验医学杂志

ABSTRACT

Objective To establish a cell level-based negative enrichmen technique to detect circulating tumor cells (CTCs) in the peripheral blood of patients with epithelial ovarian cancer.Methods The colon cancer SKOV-3 cells were mixed with 2 ml whole blood from healthy donors at different ratio.Quantification of CTCs was performed using immunomagnetic bead based negative enrichment combined with immunofluorescence antibody method.The method was evaluated the recovery rate of target cells,Samples of 32 patients with ovarian cancer and 10 controls were assayed for CTCs detection by above method.Results ①The recovery rate was ranging from 64％ ～80％ by spiking varying numbers of SKOV-3 into 2 ml blood samples of healthy volunteers.Regression analysis of number of recovered SKOV-3 cells yielded a regression equation of Y=0.782X-1.408 and a coefficient of determination of R2 =0.998.②Did not detect CK8/18-+ circulating tumor cells in 10 controls,and CK8/18+ circulating tumor cells in 18 cases of 32 Patients with ovarian cancer.The positive rate of CK 8/18 + circulating tumor cells was significantly differences between the two groups (x2 =7.681,P＜0.01).③The presence ofCTCs was significantly correlated with distant metastasis (x2 =5.776,P＜0.05).Conclusion The method of immunomagnetic bead based negative enrichment combined with immunofluorescence antibody technique for CTCs detection in peripheral blood of patients with ovarian cancer has a clinical value of application and extension.