Introduction

The Archer® FusionPlex® Heme v2 Kit is an RNA-based targeted sequencing assay that detects and identifies fusions of 87 genes associated with hematological malignancies. Using Archer’s proprietary Anchored Multiplex PCR™-based enrichment, fusions of all genes in this kit can be identified in a single sequencing assay, even without prior knowledge of fusion partners or breakpoints. Additionally, select genes are targeted for molecular barcode-enabled relative expression level detection and point mutation coverage.

For Research Use Only. Not for use in diagnostic procedures.

Highlights

  • Comprehensive fusion panel for myeloid and lymphoid origin malignancies.
  • Relevant point mutations are built into the panel to maximize the information obtained from your RNA sample
  • Molecular barcode-enabled relative expression level detection in select genes
  • Expression imbalance confirmation for most common fusion verification
  • Wide sample type compatibility, including FFPE, blood and bone marrow.



Panel Targets

FusionPlex Heme v2 Panel targets

  •  Fusion, splicing or exon skipping
  •  Mutation
  •  Expression
  •  Expression Imbalance
ABL1
 
CEBPE
   
GLIS2
   
MYC
  
PTK2B
   
ABL2
   
CEBPG
  
ID4
   
MYH11
  
RARA
 
ALK
  
CHD1
   
IKZF1
   
NF1
   
RBM15
   
BCL11B
   
CHIC2
   
IKZF2
   
NFKB2
   
ROS1
  
BCL2
 
CIITA
   
IKZF3
  
NOTCH1
  
RUNX1
  
BCL3
   
CREBBP
  
IRF4
   
NTRK3
  
RUNX1T1
  
BCL6
  
CRLF2
 
IRF8
   
NUP214
   
SEMA6A
   
BCR
   
CSF1R
   
JAK2
  
NUP98
   
SETD2
   
BIRC3
  
CTLA4
   
KAT6A
   
P2RY8
   
STIL
   
CBFB
   
DEK
   
KLF2
   
PAG1
   
TAL1
  
CCND1
 
DUSP22
   
KMT2A
   
PAX5
  
TCF3
   
CCND2
   
EBF1
   
MALT1
   
PDCD1
   
TFG
   
CCND3
  
EIF4A1
   
MECOM
  
PDCD1LG2
  
TP63
   
CD274
   
EPOR
   
MKL1
   
PDGFRA
  
TYK2
   
CDK6
   
ERG
   
MLF1
   
PDGFRB
   
ZCCHC7
   
CDKN2A
   
ETV6
  
MLLT10
   
PICALM
   
CEBPA
  
FGFR1
   
MLLT4
   
PML
  
CEBPD
   
FOXP1
   
MUC1
   
PRDM16
   

Note: Fusions involving BCR and TCR loci, including IGH, IGL and IGK, are targeted for expression and may not be explicitly called as a fusion because these often do not result in chimeric transcripts.


Related Topics



Superior panel performance

607

# of GSP2s

>99%

% Bases at 50x unique molecule coverage

>99%

Coverage uniformity > 20% mean

2.5 hours

Hands-on time

50-200ng

Input RNA required

2 Million

Recommended # of reads

9.5 hours

Time required

>90%

Unique moleculer on-target

Illumina® or Ion Torrent™

Platform

Blood, fresh frozen, FFPE

Sample type

As observed on multiple lots using 50ng gDNA input. DNA input allows for a better measure of the behavior across primers, and can be be used to assess coverage uniformity. RNA can vary by due to expression levels.




Heme v2 covers all available FISH probes

Fluorescence in site hybridization (FISH) is most often used to detect gene fusions in hematologic malignancies. To detect fusions by FISH, probes are designed to specifically detect both fusion partners if the fusion partner is known, or a breakapart FISH probe is used to detect fusion events without knowledge of the fusion partner. AMP-based NGS not only detects known and novel fusions but also identifies previously unknown fusion partner genes, information which cannot be obtained with FISH. The FusionPlex® Heme v2 kit contains primers to detect all available FISH probes, enabling detection of these important fusions as well as sequence identification of partner genes.

Disease FISH Probe Heme v2 Target
ALL BCR/ABL t(9;22)
MLL 11q23
ETV6/RUNX1 (also known as TEL-AML1)
c-MYC 8q24
PAX5 breakapart
TAL1
TCF3/PBX1
Lymphoma ALK (2q23)
BCL6 (3q27)
MYC (8q24)
CCND1/IgH t(11;24)
IgH/BCL2 t(14;18)
MALT1 (18q21)
MPN FIP1K1/PDGFRB
FGFR breakapart
MLL 11q23
PDGFRB
Disease FISH Probe Heme v2 Target
AML PML/RARA
RUNX1T1/RUNX1
CBFB breakapart
KMT2A
RPN1/MECOM
MYH11/CBFB inv(16)
DEK/NUP214 t(6;9)
MYST3/CREBBP t(8;16)
MLF1/NPM1 t(3;5)(q25.32;q35.1)
RBM15/MKL1 t(1;22)
BCR/ABL t(9;22)
NUP98 breakapart
PDGFRB
CLL CCND1/IGH t(11;14)
CML BCR/ABL t(9;22)
MYC8q24


Key fusions are covered from both sides

Reads originating from unidirectional gene-specific primers (GSPs) result in open-ended capture of gene fusions. For clinically relevant known fusion genes, GSPs are designed to capture the fusion event from both ends, enabling independent detection of a single fusion event in a single assay, thus providing internal, orthogonal verification of these important fusions. In the example shown above, 409 unique reads originating from ABL1 GSPs and 329 unique reads originating from BCR independently detected a BCR-ABL1 fusion. Furthermore, digital read counting of molecular barcodes (MBCs) ligated prior to amplification can be used to assess expression changes across the fusion gene to detect expression imbalance, providing a third internal confirmation of the detected fusion.

  • Provides independent verification of fusion event
  • + Expression imbalance over key events gives 3rd event verification

BCR:ABL1 fusions identified


How to contact us

Address

2477 55th Street, Suite 202

Boulder, CO 80301

Phone

Phone: (877) 771 1093

Phone: (303) 357 9001

All content © 2017 ArcherDX, Inc.

For Research Use Only. Not for use in diagnostic procedures. For Research Use Only. Not for use in diagnostic procedures.