TY - JOUR
T1 - Standardization of the immunocytochemical detection of neuroblastoma cells in bone marrow
AU - Swerts, Katrien
AU - Ambros, Peter F.
AU - Brouzes, Chantal
AU - Fernandez Navarro, José M.
AU - Gross, Nicole
AU - Rampling, Dyanne
AU - Schumacher-Kuckelkorn, Roswitha
AU - Sementa, Angela R.
AU - Ladenstein, Ruth
AU - Beiske, Klaus
PY - 2005/12
Y1 - 2005/12
N2 - Standard cytomorphological examination of bone marrow (BM) aspirates does not appear to be sensitive enough to detect single neuroblastoma cells. The SIOPEN Neuroblastoma Bone Marrow Committee developed a sensitive and reproducible anti-GD2 immunocytochemical assay and introduced morphological and immunocytological criteria for the interpretation of results. Fixed cytospins were incubated with a commercially available anti-GD2 monoclonal antibody and an APAAP kit. Cells fulfilling all morphological and immunocytological criteria were called criteria-positive cells (CPCs). Not convincingly interpretable cells fulfilled some, but not all, criteria, and negative cells displayed only exclusion criteria. The genetic profile of doubtful cells was checked by fluorescence in situ hybridization. Ideally, 3 × 106 cells were analyzed to reach a 95% probability of detecting one tumor cell in 1 × 106 mononuclear cells. Four quality control rounds were organized to validate the method. A total of 111 quality control samples were analyzed. Two main improvements were achieved: in discordant cases, the range between the lowest and highest reported result was reduced by half, and discordant results were only found in samples with less than 10 CPCs per 1 × 106. This article describes the first internationally standardized protocol to detect and quantify rare neuroblastoma cells by immunocytochemistry. This method is an indispensable tool for multicenter studies evaluating the clinical significance of minimal residual disease in neuroblastoma.
AB - Standard cytomorphological examination of bone marrow (BM) aspirates does not appear to be sensitive enough to detect single neuroblastoma cells. The SIOPEN Neuroblastoma Bone Marrow Committee developed a sensitive and reproducible anti-GD2 immunocytochemical assay and introduced morphological and immunocytological criteria for the interpretation of results. Fixed cytospins were incubated with a commercially available anti-GD2 monoclonal antibody and an APAAP kit. Cells fulfilling all morphological and immunocytological criteria were called criteria-positive cells (CPCs). Not convincingly interpretable cells fulfilled some, but not all, criteria, and negative cells displayed only exclusion criteria. The genetic profile of doubtful cells was checked by fluorescence in situ hybridization. Ideally, 3 × 106 cells were analyzed to reach a 95% probability of detecting one tumor cell in 1 × 106 mononuclear cells. Four quality control rounds were organized to validate the method. A total of 111 quality control samples were analyzed. Two main improvements were achieved: in discordant cases, the range between the lowest and highest reported result was reduced by half, and discordant results were only found in samples with less than 10 CPCs per 1 × 106. This article describes the first internationally standardized protocol to detect and quantify rare neuroblastoma cells by immunocytochemistry. This method is an indispensable tool for multicenter studies evaluating the clinical significance of minimal residual disease in neuroblastoma.
KW - Bone marrow
KW - Immunocytochemistry
KW - Minimal residual disease
KW - Neuroblastoma
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U2 - 10.1369/jhc.5C6661.2005
DO - 10.1369/jhc.5C6661.2005
M3 - Article
C2 - 15956022
AN - SCOPUS:23944435314
VL - 53
SP - 1433
EP - 1440
JO - Journal of Histochemistry and Cytochemistry
JF - Journal of Histochemistry and Cytochemistry
SN - 0022-1554
IS - 12
ER -