Flow cytometry and fluorescence in situ hybridization to detect residual neuroblastoma cells in bone marrow

Background In patients with neuroblastoma morphological assessment of BM for residual NB cells is not precise, particularly when the number of tumor cells is small. Procedure To develop a sensitive and rapid method of detecting NB cells in BM, we assessed the efficiency of flow cytometry (FCM) using...

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Published in:Pediatric Blood & Cancer 2005-11, Vol.45 (6), p.787-795
Main Authors: Okcu, Mehmet Fatih, Wang, Rui-Yu, Bueso-Ramos, Carlos, Schober, Wendy, Weidner, Douglas, Andrassy, Richard, Blakely, Martin, Russell, Heidi, Ozkan, Alp, Kuttesch, John, Andreeff, Michael, Chan, Ka Wah, Ater, Joann
Format: Article
Language:English
Subjects:
Adolescent
Adult
Antigens, CD - analysis
Biological and medical sciences
Bone Marrow - pathology
Bone Marrow Examination
Child
Child, Preschool
Chromosome Aberrations
Chromosomes, Human, Pair 17
Female
flow cytometry
Flow Cytometry - methods
Flow Cytometry - standards
General aspects
Humans
Immunophenotyping
In Situ Hybridization, Fluorescence - methods
In Situ Hybridization, Fluorescence - standards
Infant
Male
Medical sciences
Neoplasm, Residual - diagnosis
neuroblastoma
Neuroblastoma - diagnosis
Neuroblastoma - pathology
residual disease
Sensitivity and Specificity
Tumors
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Summary:Background In patients with neuroblastoma morphological assessment of BM for residual NB cells is not precise, particularly when the number of tumor cells is small. Procedure To develop a sensitive and rapid method of detecting NB cells in BM, we assessed the efficiency of flow cytometry (FCM) using markers CD9, CD56, and CD45. The percent of CD9+/CD56+/CD45− (NB phenotype) cells was determined by FCM in 41 samples (16 patients) at various time points. For confirmation fluorescence in situ hybridization (FISH) for 17q gain was performed. Results Nineteen of the 22 (86%) samples that were negative by morphology were positive by FCM (>0.006% CD9+/CD56+/CD45− cells). The longest time to complete the FCM study was 3 hr. In six FISH experiments the sorted CD9+/CD56+/CD45− population had a higher percentage of cells with 17q gain (11.5–95%) compared to a CD56−/CD45+ internal control population (2–8%). Conclusions Our preliminary results suggest that FCM determination of the percent of CD9+/CD56+/CD45− cells is an effective method of rapidly detecting NB cells in BM. © 2005 Wiley‐Liss, Inc.
ISSN:1545-5009
1545-5017
1096-911X
DOI:10.1002/pbc.20428