Signature®LTx Leukemia Translocation Panel v2.0
4 Diseases • 9 Translocations • 12 Fusion Transcripts • 1 Well
Download a Signature LTx v2.0 bsrochure
Use Signature® LTx v2.0 for the rapid detection and identification of the fusion transcripts from common chromosome translocations/ abnormalities associated with CML, ALL, APL, and AML. Signature® LTx v2.0 utilizes both multiplex RT-PCR and automated multiplex detection on the Luminex® 100 IS or 200™ System.
Signature® LTx v2.0 provides comprehensive information in a single well reducing the need for and cost of maintaining and training on numerous platforms and assays.
Signature® LTx v2.0 has a simple workflow that requires minimal hands-on time. The assay contains a single transfer step of amplified products directly to the bead mix, without intervening purifications. The average time to results is approximately 5 hours for 1 to 96 samples.
This data represents the relative analytical sensitivity achieved for CML Fusion Transcripts.
The analytical sensitivity of ≥1:1000 was determined by analyzing 10-fold serial dilutions of target RNA from translocation-positive cell lines into background (target negative) RNA from HL-60 cells (400 ng total RNA). Analysis of these samples showed that b2a2 and b3a2 could be reliably identified when they accounted for 0.1% of the input RNA. In addition, the pooled RNA control contains RNAs from two cell lines BV-173 (b2a2) and K-562 (b3a2). Both targets (b2a2, b3a2) and GAPDH (endogenous control) were simultaneously amplified and detected in a single well.
Signature® LTx* v2.0 Assay for Simultaneous Detection of 12 Leukemia-associated Fusion Transcripts and Endogenous Control GAPDH.
All fusion transcripts were successfully detected using the described assay. Multiplex RT-PCR was performed using 400 ng of cell line total RNAs (for 9 markers) or 10 fg of IVT RNA controls (for 12 markers) as input. Hybridization of the PCR products to the capture probes on the beads and signal detection were carried out using the Luminex System (see Methods section for details). HL-60 cell line mimics a translocation-negative sample. Non-template control (NTC) is reagent-negative control. GAPDH control, as an endogenous control, was co-amplified/detected (attenuated) along with the fusion transcripts. MLL/AF4 capture probe can detect both 10/e4 and e9/e5 fusion transcripts. Qualitative results are measured by MFI (mean fluorescent intensity). MFI values greater than or equal to 400 MFI signify positive detection for a specific translocation. A sample is considered negative if the MFI value is below 400 MFI and the GAPDH signal is greater than or equal to 400 MFI.
Attributes for Signature® LTx* v2.0:
References
Gabert J, Beillard E, van der Velden VH, Bi W, Grimwade D, Pallisgaard N, Barbany G, Cazzaniga G, Cayuela JM, Cave H, Pane F, Aerts JL, De Micheli D, Thirion X, Pradel V, Gonzalez M, Viehmann S, Malec M, Saglio G, van Dongen JJ. Standardization and Quality Control Studies of “Real-Time” Quantitative reverse Transcriptase Polymerase Chain Reaction of Fusion Gene Transcripts for Residual Disease Detection in Leukemia – A Europe Against Cancer Program. Leukemia. 17(12):2318-57. 2003.
Salto-Tellez M, Shelat SG, Benoit B, Rennert H, Carroll M, Leonard DG, Nowell P, Bagg A. Multiplex RT-PCR for the Detection of Leukemia-Associated Translocations: Validation and Application to Routine Molecular Diagnostic Practice. J Mol Diagn. 5(4):231-6. 2003.
Bock O, Reising D, Kreipe H. Multipilex RT-PCR for the Detection of Common BCR-ABL Fusion Transcripts in Paraffin-Embedded Tissues from Patients with Chronic Myeloid Leukemia and Acute Lymphoblastic Leukemia. Diagn Mol Pathol. 12(3):119-23. 2003.
