1. Real-time quantitation of minimal residual disease in inv(16)-positive acute myeloid leukemia may indicate risk for clinical relapse and may identify patients in a curable state
by Silvia Buonamici, Emanuela Ottaviani, Nicoletta Testoni, Vittorio Montefusco, Giuseppe Visani, Francesca Bonifazi, Marilina Amabile, Carolina Terragna, Deborah Ruggeri, Pier Paolo Piccaluga, Alessandro Isidori, Michele Malagola, Michele Baccarani, Sante Tura, and Giovanni Martinelli
Blood
Volume 99(2):
January 15, 2002
©2002 by American Society of Hematology

2. Position of primers and probe for CBFβ-MYH11 transcript types A, D, and E, and for the ABL gene.
Position of primers and probe for CBFβ-MYH11 transcript types A, D, and E, and for the ABL gene. CBFβ-MYH11 transcript types A, D, and E: the probe (▬) inv(16) Probe (5′ TTC AAA TTC GCG TGT CCT TCT CCG A) and the sense primer (➡) inv(16) FW (5′ TTA GCA CAA CAG GCC TTT GAA) are positioned in exon 5 of the CBFβ gene. The reverse primers ( )inv(16)A RW (5′ CAG GGC CCG CTT GGA), inv(16)D RW(5′ CCT CGG CCT CGT TAA GCA T), and inv(16)E RW (5′ GCG TCT GCT TAT TCT TGT CTA GGT T) are positioned in exons 12, 8, and 7 of theMYH11 gene for transcript types A, D, and E, respectively.ABL gene: the probe (▬) ABL Probe (5′ CCA GTA GCA TCT GAC TTT GAG CCT CAG GG) and the reverse primer ( ) ABL RW (5′ TGG GTC CAG CGA GAA GGT T) are positioned in exon 2 of the ABL gene. The sense primer (➡)ABL FW (5′ TCC TCC AGC TGT TAT CTG GAA GA) is positioned in exon 1b of the ABL gene.
Silvia Buonamici et al. Blood 2002;99:
©2002 by American Society of Hematology

3. CBFβ-MYHII amplification plot and standard curve
CBFβ-MYHII amplification plot and standard curve.(A) Amplification plot of a serial dilution of the CBFβ-MYH11 plasmid and (B) standard curve of CBFβ-MYH11 cDNA plasmid dilution for real-time RT-PCR.
CBFβ-MYHII amplification plot and standard curve.(A) Amplification plot of a serial dilution of the CBFβ-MYH11 plasmid and (B) standard curve of CBFβ-MYH11 cDNA plasmid dilution for real-time RT-PCR. Amplification plot shows the increase of reporter fluorescence during amplification and CT. The CT value decrease was proportional to the increase of the target molecules. The standard curve shows the linear correlation between the CT value and the initial copy number of the CBFβ-MYH11 and can then be used to calculate the absolute CBFβ-MYH11 fusion transcript copy number in an unknown patient sample.
Silvia Buonamici et al. Blood 2002;99:
©2002 by American Society of Hematology

4. Overall survival and disease-free survival of 18 patients with inv(16) AML.
Silvia Buonamici et al. Blood 2002;99:
©2002 by American Society of Hematology

5. Schematic representations of karyotypic and qualitative RT-PCR and real-time RT-PCR follow-up of patients with inv(16) AML.(A) Karyotypic and qualitative RT-PCR follow-up of 21 patients: karyotypic analysis negative (▵) or positive (▴) for inv(16); molecula...
Schematic representations of karyotypic and qualitative RT-PCR and real-time RT-PCR follow-up of patients with inv(16) AML.(A) Karyotypic and qualitative RT-PCR follow-up of 21 patients: karyotypic analysis negative (▵) or positive (▴) for inv(16); molecular analysis negative (○) or positive (●) for CBFβ-MYH11. (B) Real-time RT-PCR of 18 patients (patients 1, 10, 18 had insufficient material for analysis): samples taken at diagnosis or at the moment or relapse (●); samples taken at any time during or after treatment before relapse ( ); samples taken during follow-up when no subsequent relapse was recorded (○). *Samples that fell in the gray zone with CBFβ-MYH11/ABL ratios between 0.12% and 0.25%. Auto-BMT indicates autologous bone marrow transplantation; allo-BMT, allogeneic bone marrow transplantation; and PBSCT, autologous transplantation with peripheral blood stem cells.
Silvia Buonamici et al. Blood 2002;99:
©2002 by American Society of Hematology

6. CBFβ-MYH11/ABL ratios, as evaluated by real-time RT-PCR
CBFβ-MYH11/ABL ratios, as evaluated by real-time RT-PCR.One hundred twenty-five samples were taken from 21 patients with inv(16) AML during or after treatment in the absence of subsequent relapse (Group A), at any time during follow-up before relapse (Group...
CBFβ-MYH11/ABL ratios, as evaluated by real-time RT-PCR.One hundred twenty-five samples were taken from 21 patients with inv(16) AML during or after treatment in the absence of subsequent relapse (Group A), at any time during follow-up before relapse (Group B), or at the time of diagnosis or relapse (Group C). Differences among the 3 groups were all highly significant (P < .0001), as determined by the Kruskal-Wallis test. When values were lower than 0.12%, no subsequent relapse was recorded, but when values were greater than 0.25%, relapse always occurred. Values fell within the intermediate gray zone for 6 samples. Median values of each group (—).
Silvia Buonamici et al. Blood 2002;99:
©2002 by American Society of Hematology