b'Gel & Capillary | RUO AssaysGel and CapillaryPML/RARa t(9;22) Translocation Assays Research Use Only (RUO) AssayPML/RARa t(15;17) Translocation AssaysAssay Use cytogenetic or molecular detection of these t(15;17) translocations. 4Three PML/RARA translocation patterns have been identified: PML/RAR t(15;17) Translocation Assays are useful for studies involving: Type A is the short form (S-form); the breakpoint occurs within the Identification of acute promyelocytic leukemia (APL) breakpoint cluster region 3 (Bcr3). Type B is the long form (L-form) and the breakpoint occurs within Bcr1. There is a third type B variant Monitoring and evaluation of disease recurrence or variable form (V-form) whose breakpoint occurs within Bcr2. Detection and assessment of residual disease Identification of the PML-RARA translocation is important in APL because it is correlated with responsiveness to treatment with all-Evaluation of new research and methods in malignancy studies trans retinoic acid (ATRA). Patients incorrectly diagnosed with APL by clinical and morphologic criteria alone are typically unresponsive to treatment with ATRA. APL patients are also at risk for disseminated Summary and Explanation of the Testintravascular coagulation (DIC), which can become more severe Four master mixes are included in these assay kits. Master mixesduring conventional chemotherapy. As a result, there is clearly a need are used to amplify complementary DNA (cDNA) produced fromfor a rapid, sensitive, and reliable molecular assay that identifies PML-specimen(s), as well as positive and negative RNA controls (included).RARA transcripts associated with APL. This Assay uses both non-nested Primers target an internal control transcript (RARA, formerly known(master mix 1) and nested (master mixes 2a, 2b, and 2c) reverse as RAR) and the variety of Bcr1, Bcr2, and Bcr3 type transcriptstranscriptase PCR for faster and significantly more sensitive results expressed from PML-RARA translocations. The limit of detection forthan cytogenetics or other methods.this assay is one PML-RARA positive cell in a background of one hundred thousand normal cells. Amplicon products can be analyzed by differential fluorescence detection using capillary electrophoresisSpecimen Requirementsor standard gel electrophoresis. A PML-RARA translocation is5 mL of peripheral blood, bone marrow biopsy, or bone marrow indicated if just one of the 2nd round master mixes (Mix 2b or Mix 2c) generates product(s) of the valid size. Reagents for RNA extractionaspirate anticoagulated with heparin or EDTA; or,and reverse transcription are not included. This assay is compatibleArchived cells frozen in 10% DMSO + 90% Fetal Bovine with all standard RNA extraction and cDNA synthesis methods. This isSerum (FBS).a qualitative assay and has not been validated for quantitative use.ReferenceBackground1.H De Th et al., Nature 347: 558-561 (1990).Acute promyelocytic (M3) leukemia (APL) is a distinct form of acute2. H De Th et al., Cell 66: 675-684 (1991).myeloid leukemia (AML) representing approximately 10% of AMLs.3. A Kakizuka et al., Cell 66: 663-674 (1991).These leukemias often express PML-RARA transcripts from t(15;17) chromosomal translocations that fuse the PML (or MYL) gene4. WH Miller et al., Proc. Natl. Acad. Sci. 89: 2694-2698 (1992).on chromosome 15 with the retinoic acid receptor a (RARA) gene on chromosome 17. 1,2,34Diagnosis of APL is typically based upon identification of promyelocytes with distinctive morphology plus Figure Legend: This figure shows the genomic organization of the PML andPML GeneRARA genes on chromosomes 15 and 17, respectively. Boxes represent exonChromosome 15regions of the PML (red boxes) and RARA (orange) encoding exons. The solid black line represents intron regions, which were left incompletely spliced toRARA Geneassist in demarcation of the exon segments. Primers are indicated by arrows,Chromosome 17and the size of several of the products are indicated below the translocated gene segments. S-form (Bcr3) and L-form (Bcr1) PML-RARA translocations are depicted in the lower portion of the figure.126'