Immunobead-PCR: a technique for the detection of circulating tumor cells using immunomagnetic beads and the polymerase chain reaction.
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.
Hardingham
J E
JE
Haematology-Oncology Department, Queen Elizabeth Hospital, Woodville, Australia.
Kotasek
D
D
Farmer
B
B
Butler
R N
RN
Mi
J X
JX
Sage
R E
RE
Dobrovic
A
A
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