Quantitative Analysis of AMLI-ETO Fusion Transcripts in t(8;21} Positive AML Using Real-Time RT-PCR m'I
Results
To access the sensitivity of our real-time peR protocol we performed a limiting
dilution series of a single Kasumi 1 cell in the background of 10, 100, 1000, 10,000,
100,0001,000,000 normal human cells. This revealed a sensitivity of 10-s (1 Kasumi 1 cell in the background of 100,000 normal cells was detectable) (Fig. 2).
Repeated standard curve generation using serial dilutions of Kasumi 1 cells (lOS,
104, 103, 10 2 ) was carried out. peR efficiencies for the amplification of AMLl-ETO
and G6PDH were calculated using the following formula:
peR efficiency = lO-lfslope
0.9
0.8
0.7
0.3
0.2
01
o
.0.1 I
o
I
5
T"get AMl1·ETO
I
10
1:10
1:100
1:1000
"10000
1;100000
I
15
I
I
I
I
20
25
3)
l5
CJ,de NIIIIber
I
40
I
45
I
50
Fig. 2. AMLl-ETO transcripts of 1 t(8;21) positive Kasumi 1 cell in the background of 10,100,
1000, 10,000 and 100,000 normal human cells were detected
Sensitivity of
Real-Time RT-PCR
Protocol
PCR Efficiency