Detection of isolated tumor cells in peripheral blood and in BM: evaluation of a new enrichment method.
Cell enrichment methods that deal with larger volumes of peripheral blood and BM are needed for increased sensitivity of detection, characterization and quantification of isolated tumor cells (ITC). This study was designed to evaluate a new procedure, the RosetteSep-Applied Imaging Rare Event (RARE) detection method, which depletes the majority of the erythrocytes and leucocytes in a peripheral blood (PB) sample, thereby negatively enriching tumor cells if present. This enrichment procedure allows for increased sensitivity, by analyzing a 5-10 fold larger volume of blood, compared with a direct immunocytochemical (ICC) technique, with minimal impact on laboratory workload. Model experiments showed comparable tumor cell recoveries between the two tested methods, both in PB and BM. Clinical samples were evaluated using paired PB and BM samples from 95 carcinoma patients. Analysis of PB results showed that 25.3% had > or = 1 tumor cell detected by the RARE procedure, compared with 5.2% after direct ICC analysis, analyzing a 10-fold larger volume by the RARE procedure. The direct ICC analysis of BM from the same patients revealed 16.8% positive. The ITC detection differed both quantitatively and qualitatively between BM and PB, as samples with high numbers of ITC in BM were still negative in PB. The clinical significance of ITC in blood still needs to be established. However, the easy access of peripheral blood, and the increased sensitivity obtained by increasing the sample volume with the RARE procedure, suggests that the value of peripheral blood analysis should be tested in parallel in studies where ITC detection in BM is performed.
Naume
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B
Department of Oncology, The Norwegian Radium Hospital, University of Oslo, Montebello, Oslo, Norway.
Borgen
E
E
Tøssvik
S
S
Pavlak
N
N
Oates
D
D
Nesland
J M
JM
D000911:Antibodies, Monoclonal; D001853:Bone Marrow / Q000473:pathology*; D001856:Bone Marrow Examination; D019046:Bone Marrow Neoplasms / Q000175:diagnosis* / Q000556:secondary; D002452:Cell Count / Q000295:instrumentation / Q000379:methods; D045744:Cell Line, Tumor; D002499:Centrifugation, Density Gradient; D004912:Erythrocytes / Q000166:cytology / Q000276:immunology; D005260:Female; D006801:Humans; D019044:Image Cytometry / Q000379:methods; D007150:Immunohistochemistry / Q000379:methods*; D018189:Immunomagnetic Separation / Q000379:methods*; D007962:Leukocytes / Q000166:cytology / Q000276:immunology; D008297:Male; D009369:Neoplasms / Q000097:blood* / Q000175:diagnosis*; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D011237:Predictive Value of Tests; D015203:Reproducibility of Results
D000911:Antibodies, Monoclonal