[Multicentric evaluation of MDR phenotype in leukemia: intermediate analysis of the French study].
Thirty-six French centres are involved in an evaluation of the techniques used for MDR phenotype measurement. Until now, 14 samples of various kinds of leukemia (mainly acute myelogenous leukemia) and three cell lines with different levels of resistance were sent by one centre and tested. MRK16 antibody was used for flow cytometry and immunocytochemistry, RNA was measured by RT-PCR, rhodamine or anthracyclin efflux were tested for functional assay. Wide discrepancies were observed in the results, mainly with flow cytometry, specially for the samples with a probable low level of MDR1 expression. The importance of histogram interpretation was documented by the comparative analysis of results obtained on cells already marked with MRK16, fixed and sent to all centers. The use of the ratio of the mean of fluorescence, instead of percentage, should help for standardization. The use of only one control RNA (used at different dilutions) for standardisation of RT-PCR could help in decreasing the discrepancies observed. The mean of fluorescence should also be used for expressing the rhodamine cell content.
Perrot
J Y
JY
Service d'hématologie biologique, Hôtel-Dieu, Paris, France.
Faussat
A M
AM
Zhou
D C
DC
Zittoun
R
R
Robert
J
J
Marie
J P
JP
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics; D015152:Blotting, Northern; D018432:Drug Resistance, Multiple / Q000235:genetics*; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D007938:Leukemia / Q000235:genetics*; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012235:Rhodamines / Q000378:metabolism; D012680:Sensitivity and Specificity
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D012235:Rhodamines