BCR-ABL P190 (mbcr) One-Step Detection Kit
BCR-ABL and Cancers Chromosomal translocations involving BCR on chromosome 22 and ABL on chromosome 9 are the most frequent aberrations found in patients with Acute Lymphoblastic Leukemia (ALL). This rearrangement, also called the Philadelphia Chromosome (Ph) is present in approximately 30% of adult ALL patients (> 50% in patients over 50 years of age).
The breakpoint on chromosome 22 occurs in the first intron of the BCR gene (also known as minor breakpoint cluster region; mbcr), and the breakpoint on chromosome 9 occurs between exons 1 and 2 of the ABL gene (e1a2). This joins exon 1 of BCR to exon 2 of ABL, creating a 190 kDa fusion protein with a constitutively active kinase domain.
Tyrosine kinase inhibitors, such as STI-571 (imatinib; IM, Gleevec®), have been shown to greatly improve the survival of patients with ALL. Current treatment guidelines recommend monitoring the fusion:reference gene ratios in patients undergoing therapy.
One-Step Quantitative RT-PCR Current guidelines from Europe Against Cancer (EAC) program and National Comprehensive Cancer Network (NCCN) recommend the use of quantitative real-time PCR to detect and quantify BCR-ABL expression before, during, and after IM treatment to assess the minimal residual disease (MRD) in patients diagnosed with ALL. This assay is designed to conform to those guidelines.
Our BCR-ABL P190 (mbcr) One-Step Detection Kit has been formulated for highly reproducible first-strand cDNA synthesis and subsequent real-time PCR in a single tube. A combination of validated primers, hydrolysis probes, and hot-start DNA polymerase ensures that the BCR-ABL P190 (mbcr) One-Step Detection Kit leads to highly-specific and ultra-sensitive results in a single step. It uses the Comparative Ct method (Pfaffl method) to determine fusion:reference gene ratios.
Assay Features The BCR-ABL P190 (mbcr) One-Step Detection Kit provides reagents to detect and quantify the e1a2 fusion gene transcript using total RNA isolated from blood or bone marrow. Some of the benefits of using this kit are:
• One-step cDNA synthesis and quantification – no need to perform separate cDNA synthesis and qPCR • Low contamination risk – no post-PCR product handling • High-throughput – can screen up to 46 samples in a single 96-well run • Fast – results in under 80 minutes after PCR start • Accurate – reference and target genes assessed in the same reaction
Equipment and Materials The BCR-ABL P190 (mbcr) One-Step Detection Kit requires a real-time PCR instrument capable of detecting FAM™ and VIC® fluores. It does not include reagents and columns for isolation of total RNA from blood or bone marrow.
Intended Use The BCR-ABL P190 (mbcr) One-Step Detection Kit is for Research Use Only (RUO) in the US and for In Vitro Diagnostic Use (CE- IVD) in Europe.
Ordering Information Product Name Cat. No. BCR -ABL P190 (m bcr) One -Step Detection Kit – 46 samples BCR 190 -QRT46
Gleevec® is a registered trademark of Novartis, FAM™ and VIC® are registered trademarks of Applera Corporation or its subsidiaries.
BÜHLMANN Laboratories • Baselstrasse 55 • CH -4124 Schönenbuch/Basel • Switzerland Phone +41 61 487 12 12 • Fax +41 61 487 12 34 • [email protected] • www.buhlmannlabs.ch