Sensitivity and applicability of different methods for detection of terminal transferase in leukemia.
The sensitivity of terminal deoxynucleotidyl transferase (TdT) assay methods was examined by using a mixture of the TdT-positive lymphoblastic leukemia cell line NALM-18 and the TdT-negative erythroleukemia cell line K-562. The biochemical assay could detect TdT activity in the mixture containing NALM-18 cells at concentrations of more than 10 percent. The immunofluorescent (IF) method could detect positive cells in the mixture containing NALM-18 cells at concentrations of more than 1 percent. Furthermore, an approximately 10(5)-fold increase in sensitivity was obtained by the combination of RT-PCR and subsequent Southern blotting, as compared to biochemical assay. In many leukemia cases the expression of TdT-mRNA corresponded well to that of TdT protein. However, in some patients with leukemia, only TdT-mRNA was detectable by RT-PCR without any expression of TdT protein. A PCR-based technique enables us to detect TdT transcripts at the highest sensitivity, but does not allow the characterization of each positive cell. IF analysis is simple and sensitive, but may sometimes cause nonspecific reactions. All these techniques have some advantages and some faults, therefore, the results obtained from clinical studies using these techniques should be interpreted with caution.
Sasaki
R
R
Department of Hematology, Jichi Medical School, Tochigi, Japan.
Fukushima
M
M
Miura
Y
Y
Chang
L M
LM
Bollum
F J
FJ
D001483:Base Sequence; D001752:Blast Crisis; D002460:Cell Line; D004253:DNA Nucleotidylexotransferase / Q000032:analysis* / Q000096:biosynthesis*; D017931:DNA Primers; D005455:Fluorescent Antibody Technique; D006801:Humans; D007938:Leukemia / Q000145:classification / Q000201:enzymology* / Q000473:pathology; D004915:Leukemia, Erythroblastic, Acute; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma; D012333:RNA, Messenger / Q000032:analysis; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D012333:RNA, Messenger; D004253:DNA Nucleotidylexotransferase