BCR-ABL One-Step Detection Kit
No. of Reactions*
|BCR-ABL P210 (Mbcr) One-Step Detection Kit
|BCR-ABL P190 (mbcr) One-Step Detection Kit
*Includes all controls
BCR-ABL and Cancers
Approximately 95% of cases of Chronic Myeloid Leukemia (CML) and about 35% of Acute Lymphoblastic Leukemia (ALL) are associated with the presence of a t(9;22)(q34;q11) chromosomal translocation (Philadelphia chromosome, Ph). This results in creation of an oncogenic fusion gene between ABL proto-oncogene and BCR on chromosomes 9 and 22, respectively. The breakpoint on chromosome 22 occurs between exons 12 and 16 of the BCR gene (also known as major breakpoint cluster region; Mbcr), while the breakpoint on chromosome 9 mostly occurs between exons 1 and 2 of the ABL gene. The two most common fusion variants are called b2a2 and b3a2, which encode for a constitutively active chimeric tyrosine kinase of 210kDa (P210).
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 inhibit the growth of tumor cells and reduce the patient’s risk of reaching “blast crisis”, the final phase of CML associated with decreased response and short survival. Complete cytogenetic response is achieved quite rapidly in CML patients treated with IM, thus a sensitive method to detect and quantify the fusion gene transcripts is required to accurately assess the response during 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 with CML and ALL. This assay is designed to conform to those guidelines.
Our BCR-ABL P210 (Mbcr) and P190 (mbcr) One-Step Detection Kits have 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 kits lead to highly-specific and ultra-sensitive results in a single step. It uses the Comparative Ct method (Pfaffl method) to determine target:reference gene ratios that are aligned with the International Scale (IS).
The BCR-ABL P210 (Mbcr) One-Step Detection Kit includes reagents to detect and quantify b2a2 and b3a2 fusion gene transcripts, while the BCR-ABL P190 (mbcr) One-Step Detection Kit includes reagents to detect and quantify e1a2 fusion gene transcripts 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
- Results aligned with IS – control standard included with the P210 kit is aligned with WHO BCR-ABL primary standards
- 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
- Multiplexed – reference and target genes assessed in the same reaction
Equipment and Materials
The BCR-ABL P210 (Mbcr) and P190 (mbcr) One-Step Detection Kits require a real-time PCR instrument capable of detecting FAM™ and VIC® fluores. They do not include reagents and columns for isolation of total RNA from blood or bone marrow.
These kits include reagents required for cDNA synthesis and PCR amplification/detection, as well as validated reaction controls. Columns and reagents for RNA isolation are not included.
Available for research use only (RUO). Not for use in diagnostic procedures.
Available for research use only (RUO) and diagnostic (CE-IVD) use.
- Analytical sensitivity for P210 at 10-4.5
- Analytical sensitivity for P190 at 10-4.0
- Results aligned with IS (P210 only)
- Requires low amount of RNA input
- Simple setup and interpretation
- Complementary EntroGen PCR Analysis Software
- Multiplexed: BCR-ABL/ABL
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