Iam AML1-ETO Kit
The faster molecular solution for one-step detection of AML1-ETO translocation
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KIT for Oncohematology Overview
Background
AML1-ETO/t (8;21)(q22;q22)/ RUNX1-RUNX1T1 is a recurrent genetic abnormality in both childhood and adult Acute Myeloid Leukemia (5-10% of all AML and nearly 40% of the AML FAB M2 subtype). Although this translocation is associated with a good prognosis, only 50% of patients are cured and its rapid molecular identification is crucial for risk stratification of patients and therapeutic decisions.
Why to choose it
Molecular identification of AML1-ETO translocations in 15 minutes directly from RNA. Effective screening of AML is now possible.
For use on
Benefits
Identification of positive samples in less than 15 min, starting from extracted RNA.
closed format
The assay works directly on 500 ng RNA with no need for multistep procedures: decreased risk of contamination and errors.
No false positive results and absence of primer dimers.
The Analytical Specificity has been established on negative cell line RNA extracted by both the Qiagen RNeasy kit and modified Trizol.
| Sample type | Total No. Replicates | % Analytical Specificity |
|---|---|---|
| Sample typeCell lines | Total No. Replicates400 | % Analytical Specificity100% |
| Sample typeNTC | Total No. Replicates210 | % Analytical Specificity100% |
| Total replicates | Total No. Replicates610 | % Analytical Specificity100% |
Internal endogenous control and quality controls are included to allow validation of results.
Detection of the AML1-ETO translocation in 10 thousand folds diluted samples in less than 20 minutes.
The test consistently detects the AML1-ETO transcript down to 10-4 dilution (Kasumi-1 in HL60) in less than 20 minutes, both on Qiagen and manual extracted RNA. Dilutions at 10-5 of Kasumi-1 in HL60, have also been revealed with a probability close to 60%.
| Extraction method | Kasumi-1 into HL60 RNA dilution | No. Replicates | Mean Tt (min) | % Detection |
|---|---|---|---|---|
| Extraction methodQiagen Rneasy Kit | Kasumi-1 into HL60 RNA dilution10³ | No. Replicates40 | Mean Tt (min)14,37 | % Detection 100% |
| Extraction methodModified TRIzol | Kasumi-1 into HL60 RNA dilution10³ | No. Replicates40 | Mean Tt (min)14,05 | % Detection 100% |
| Extraction methodQiagen Rneasy Kit | Kasumi-1 into HL60 RNA dilution104 | No. Replicates40 | Mean Tt (min)16,67 | % Detection 100% |
| Extraction methodModified TRIzol | Kasumi-1 into HL60 RNA dilution104 | No. Replicates40 | Mean Tt (min)16,94 | % Detection 100% |
| Total replicates | Kasumi-1 into HL60 RNA dilution | No. Replicates160 | Mean Tt (min) | % Detection 100% |
100% concordance with the traditional RT-PCR (Biomed Protocol*, by Van Dongen et al)
*Leukemia. 1999 Dec;13(12):1901-28.
Clinical archived RNA samples, extracted by Qiagen RNeasy kit, modified TRIzol and Maxwell, have been retrospectively tested by Q-LAMP and the results have been compared RT-PCR and found to be full concordant.
| Sample status | % Agreement with PCR (BIOMED protocol) |
|---|---|
| Sample statusAML1-ETO positive | % Agreement with PCR (BIOMED protocol)100% (N=35) |
| Sample statusAML1-ETO negative | % Agreement with PCR (BIOMED protocol)100% (N=30) |
| Total replicates | % Agreement with PCR (BIOMED protocol)100% |
Superior robustness to inhibitors, RNA degradation and suboptimal target quantities.
The intrinsic robustness of the enzyme employed in Q-LAMP results in a enhanced robustness versus the classical inhibitor factors of PCR.
vs. PCR
Q-LAMP is a very rapid, easy, accurate, convenient and robust method for detection of AML1-ETO translocations
| Q-LAMP | RT-PCR (Biomed) | |
|---|---|---|
| SubjectRNA input | Q-LAMP500 ng/rx robust down to 25 ng/rx | RT-PCR (Biomed)1μg/rx |
| SubjectSteps | Q-LAMP1 | RT-PCR (Biomed)3 |
| SubjectAssay format | Q-LAMPMULTIPLEX | RT-PCR (Biomed)SIMPLEX |
| SubjectControl of reaction | Q-LAMPINTERNAL | RT-PCR (Biomed)EXTERNAL |
| SubjectRobustness | Q-LAMP– to chemical contamination – to PCR inhibitors – to degraded samples |
RT-PCR (Biomed)To chemical contamination |
| SubjectResult interpretation | Q-LAMPOBJECTIVE | RT-PCR (Biomed)SUBJECTIVE |
| SubjectTime to result | Q-LAMP40min | RT-PCR (Biomed)3h 30min |
Libraries
Ordering info
| Product | Code | Reactions |
|---|---|---|
| Product Iam AML1-ETO |
Code V35AML | Reactions 24 |
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