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Application of immunomagnetic cell enrichment in combination with RT-PCR for the detection of rare circulating head and neck tumor cells in human peripheral blood.
- Source :
-
Cytometry. Part B, Clinical cytometry [Cytometry B Clin Cytom] 2007 Sep; Vol. 72 (5), pp. 310-23. - Publication Year :
- 2007
-
Abstract
- Detection of rare, circulating tumor cells (CTC's) in human peripheral blood is a potential indicator of prognosis and diagnosis in oncology. Typical methods to detect these CTC's are either by immunocytochemistry (ICCS) or RT-PCR. However without accurate, rapid, and reproducible enrichment processes, these detection techniques are labor intensive and/or unreliable. In this article, a repeatable enrichment process that included a flow-through immunomagnetic cell separation system, the quadrupole magnetic sorter (QMS) was optimized with the aid of a statistical analysis software package. The QMS was operated in a negative mode of operation by immunomagnetically targeting normal human peripheral blood lymphocytes (PBL) through the CD45 surface marker. Three head and neck squamous carcinoma cell lines (HNSCC), Detroit-562, SCC-4, and CAL-27, were used to determine the sensitivity of RT-PCR for the epidermal growth factor receptor (EGFR) in spiked PBL. The detection purity needed for detection was found to be one cell in 10(4), one cell in 10(3), and one cell in 10(5) for the Detroit-562, SCC-4, and CAL-27, respectively. The actual number of cancer cells needed for RT-PCR detection ranged from 30 to 1 cell. To mimic the potential concentration of rare CTC present in peripheral blood of cancer patients, the spiking concentration was chosen to be one cancer cell per 10(5) total leukocytes from healthy donors. Using a single step immunomagnetic labeling, the final, optimized enrichment process produced a 57.6 +/- 30.3-fold (n = 6) enrichment of the rare cancer cells with a final cancer cell recovery of (77.8 +/- 6.6)%.<br /> (Copyright 2007 Clinical Cytometry Society.)
- Subjects :
- Biomarkers, Tumor immunology
Cell Line, Tumor
Data Interpretation, Statistical
ErbB Receptors analysis
ErbB Receptors immunology
Head and Neck Neoplasms physiopathology
Humans
Immunomagnetic Separation instrumentation
Leukocyte Common Antigens analysis
Leukocyte Common Antigens immunology
Lymphocytes immunology
Predictive Value of Tests
Sensitivity and Specificity
Biomarkers, Tumor analysis
Head and Neck Neoplasms blood
Head and Neck Neoplasms diagnosis
Immunomagnetic Separation methods
Neoplastic Cells, Circulating pathology
Reverse Transcriptase Polymerase Chain Reaction methods
Subjects
Details
- Language :
- English
- ISSN :
- 1552-4949
- Volume :
- 72
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Cytometry. Part B, Clinical cytometry
- Publication Type :
- Academic Journal
- Accession number :
- 17205568
- Full Text :
- https://doi.org/10.1002/cyto.b.20177