title: Immunobead-PCR: a technique for the detection of circulating tumor cells using immunomagnetic beads and the polymerase chain reaction.
abstract: The presence of tumor cells in the circulation may predict disease recurrence and metastases. We have developed a sensitive technique for the detection of carcinoma cells in blood, using immunomagnetic beads to enrich for epithelial cells and the polymerase chain reaction to identify a tumor marker. The colon carcinoma cell line SW480, homozygous for a K-ras codon 12 mutation, was used to establish optimal conditions. The SW480 cells were serially diluted in normal blood and incubated with immunomagnetic beads labeled with a monoclonal antibody specific for epithelial cells. Cells bound to the beads were retrieved using a magnetic field and the presence of K-ras codon 12 mutations determined by a polymerase chain reaction based analysis. SW480 cells could be detected in dilutions up to 1 SW480 cell/10(5) leukocytes in whole blood.
authors.affiliation: Haematology-Oncology Department, Queen Elizabeth Hospital, Woodville, Australia.
authors.forename: J E
authors.initials: JE
authors.lastname: Hardingham
authors.forename: D
authors.initials: D
authors.lastname: Kotasek
authors.forename: B
authors.initials: B
authors.lastname: Farmer
authors.forename: R N
authors.initials: RN
authors.lastname: Butler
authors.forename: J X
authors.initials: JX
authors.lastname: Mi
authors.forename: R E
authors.initials: RE
authors.lastname: Sage
authors.forename: A
authors.initials: A
authors.lastname: Dobrovic
mesh_terms: D001483:Base Sequence; D015179:Colorectal Neoplasms / Q000175:diagnosis; D006801:Humans; D008280:Magnetics; D008863:Microspheres; D008969:Molecular Sequence Data; D009360:Neoplastic Cells, Circulating; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D014407:Tumor Cells, Cultured