Dry-reagent disposable dipstick test for visual screening of seven leukemia-related chromosomal translocations.
We report the first dry-reagent, disposable, dipstick test for molecular screening of seven chromosomal translocations associated with acute and chronic leukemia. The dipstick assay offers about 10 times higher detectability than agarose gel electrophoresis and, contrary to electrophoresis, allows confirmation of the sequence of the polymerase chain reaction (PCR) product by hybridization within a few minutes without the need of instrumentation. Biotinylated amplified DNA is hybridized with a dA-tailed probe and applied to the strip, which contains oligo(dT)-conjugated gold nanoparticles in dry form. Upon immersion of the strip in the appropriate buffer, the solution migrates and the hybrids are captured by immobilized streptavidin at the test zone generating a characteristic red line. The excess nanoparticles are captured by oligo(dA) strands immobilized at the control zone of the strip producing a second red line. We studied the: t(9;22)(q34;q11), t(15;17)(q22;q21), t(11;17)(q23;q21), t(5;17)(q32;q21), t(11;17)(q13;q21), t(8,21)(q22;q22) and inv(16)(p13;q22) that generate the BCR-ABL, PML-RARa, PLZF-RARa, NPM-RARa, NuMA-RARa, AML1-ETO and CBFbeta-MYH11 fusion genes, respectively. A single K562 cell was detectable amidst 10(6) normal leukocytes. A dipstick test was developed for actin, as a reference gene. The dipstick assay with appropriate probes can be used for identification of the fusion transcripts involved in the translocation.
Kalogianni
Despina P
DP
Department of Chemistry, University of Patras, Patras 26500, Greece.
Bravou
Vasiliki
V
Christopoulos
Theodore K
TK
Ioannou
Penelope C
PC
Zoumbos
Nicholas C
NC
D000208:Acute Disease; D050676:Core Binding Factor Alpha 2 Subunit / Q000235:genetics; D016044:Fusion Proteins, bcr-abl / Q000235:genetics; D006801:Humans; D007202:Indicators and Reagents; D007938:Leukemia / Q000175:diagnosis*; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000175:diagnosis; D007951:Leukemia, Myeloid / Q000175:diagnosis; D015473:Leukemia, Promyelocytic, Acute / Q000175:diagnosis; D009693:Nucleic Acid Hybridization / Q000379:methods*; D015514:Oncogene Proteins, Fusion / Q000235:genetics*; D016133:Polymerase Chain Reaction; D000075142:RUNX1 Translocation Partner 1 Protein; D015203:Reproducibility of Results; D014178:Translocation, Genetic
C107844:AML1-ETO fusion protein, human; D050676:Core Binding Factor Alpha 2 Subunit; D007202:Indicators and Reagents; D015514:Oncogene Proteins, Fusion; D000075142:RUNX1 Translocation Partner 1 Protein; C406060:abl-bcr fusion protein, human; C093476:promyelocytic leukemia-retinoic acid receptor alpha fusion oncoprotein; D016044:Fusion Proteins, bcr-abl