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Table 3 RNA-based analysis of unequivocal ALK cases showed 100% concordance between digital probe-based assay and IHC and FISH results. ALK fusion detection based on 3′/5′ imbalance MPS assay showed 25% deviation for IHC-negative/FISH-negative cases and 33% deviation for IHC-positive/FISH-positive cases. Data showed 92% concordance between ALK fusion breakpoint detection by MPS assay and IHC and FISH for IHC-negative/FISH-negative samples and 100% accordance for IHC-positive/FISH-positive samples

From: RNA-based analysis of ALK fusions in non-small cell lung cancer cases showing IHC/FISH discordance

Sample

IHC

FISH

Massive Parallel Sequencing (MPS) (IonTorrent)

Probe-Based Assay (NanoString)

Fusion

Fusion

3′/5’

Fusion

3′/5’

IHC-negative/FISH-negative

 P01

–

–

NA

NA

NA

NA

ND

 

 P02

–

–

–

–

±

–

–

–

 

 P03

–

–

–

–

±

–

–

–

 

 P04

–

–

–

–

–

–

 

 P05

–

–

–

–

–

–

 

 P06

–

–

–

–

ND

 

 P07

–

–

–

–

ND

 

 P08

–

–

–

–

ND

 

 P09

–

–

–

–

ND

 

 P10

–

–

–

–

ND

 

 P11

–

–

–

–

ND

 

 P12

–

–

–

–

ND

 

IHC-positive/FISH-positive

 P13

+

+

+

+

+

–

+

+

V1

 P14

+

+

+

±

+

+

V1

 P15

+

+

+

+

+

+

V3a/b

 P16

+

+

+

+

+

+

V2

 P17

+

+

+

+

ND

V1

 P18

+

+

+

+

+

+

ND

V6

  1. Fusion: fusion detection with amplicons/probes covering ALK fusion breakpoint; 3′/5′: fusion detection with amplicons/probes covering 3′ and 5′ end; NA: Not Analyzable; ND: Not Determined; +: fusion detected; −: fusion not detected; ±: fusion uncertain; V1: EML4_13:ALK_20; V2: EML4_20:ALK_20; V3a: EML4_6a:ALK_20; V3b: EML4_6b:ALK_20; V6: EML4_13:ALK_20_ins69