PMID- 24787965
OWN - NLM
STAT- MEDLINE
DCOM- 20150130
LR  - 20240210
IS  - 1556-1380 (Electronic)
IS  - 1556-0864 (Print)
IS  - 1556-0864 (Linking)
VI  - 9
IP  - 6
DP  - 2014 Jun
TI  - A comparison of immunohistochemical assays and FISH in detecting the ALK 
      translocation in diagnostic histological and cytological lung tumor material.
PG  - 769-74
LID - 10.1097/JTO.0000000000000157 [doi]
AB  - INTRODUCTION: Detection of the ALK rearrangement in a solid tumor gives these 
      patients the option of crizotinib as an oral form of anticancer treatment. The 
      current test of choice is fluorescence in situ hybridization (FISH), but various 
      cheaper and more convenient immunohistochemical (IHC) assays have been proposed 
      as alternatives. METHODS: Fifteen FISH-positive cases from patients, seven with 
      data on crizotinib therapy and clinical response, were evaluated for the presence 
      of ALK protein using three different commercially available antibodies: D5F3, 
      using the proprietary automated system (Ventana), ALK1 (Dako), and 5A4 (Abcam). A 
      further 14 FISH-negative and three uncertain (<15% rearrangement detected) cases 
      were also retrieved. Of the total 32 specimens, 17 were excisions and 15 were 
      computed tomography-guided biopsies or cytological specimens. All three 
      antibodies were applied to all cases. Antibodies were semiquantitatively scored 
      on intensity, and the proportion of malignant cells stained was documented. 
      Cutoffs were set by receiver operating curve analysis for positivity to optimize 
      correct classification. RESULTS: All three IHC assays were 100% specific but 
      sensitivity did vary: D5F3 86%, ALK 79%, 5A4 71%. Intensity was the most 
      discriminating measure overall, with a combination of proportion and intensity 
      not improving the test. No FISH-negative IHC-positive cases were seen. Two 
      FISH-positive cases were negative with all three IHC assays. One of these had 
      been treated with crizotinib and had failed to show clinical response. The other 
      harbored a second driving mutation in the EGFR gene. CONCLUSIONS: IHC with all 
      three antibodies is especially highly specific (100%) although variably sensitive 
      (71%-86%), specifically in cases with scanty material. D5F3 assay was most 
      sensitive in these latter cases. Occasional cases are IHC-positive but 
      FISH-negative, suggesting either inaccuracy of one assay or occasional tumors 
      with ALK rearrangement that do not express high levels of ALK protein.
FAU - Le Quesne, John
AU  - Le Quesne J
AD  - *Department of Histopathology, Royal Brompton and Harefield NHS Foundation Trust, 
      London; daggerCentre for Molecular Pathology, The Royal Marsden Hospital, Sutton, 
      Surrey; double daggerDepartment of Oncology, The Royal Marsden Hospital; and  section signDepartment of 
      Histopathology, Royal Marsden Hospital, Chelsea, London, United Kingdom.
FAU - Maurya, Manisha
AU  - Maurya M
FAU - Yancheva, Slaveya G
AU  - Yancheva SG
FAU - O'Brien, Mary
AU  - O'Brien M
FAU - Popat, Sanjay
AU  - Popat S
FAU - Wotherspoon, Andrew C
AU  - Wotherspoon AC
FAU - de Castro, David Gonzalez
AU  - de Castro DG
FAU - Nicholson, Andrew G
AU  - Nicholson AG
LA  - eng
GR  - 18824/CRUK_/Cancer Research UK/United Kingdom
GR  - MC_UP_1203/1/MRC_/Medical Research Council/United Kingdom
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Thorac Oncol
JT  - Journal of thoracic oncology : official publication of the International 
      Association for the Study of Lung Cancer
JID - 101274235
RN  - 0 (Antibodies)
RN  - 0 (Protein Kinase Inhibitors)
RN  - 0 (Pyrazoles)
RN  - 0 (Pyridines)
RN  - 53AH36668S (Crizotinib)
RN  - EC 2.7.10.1 (ALK protein, human)
RN  - EC 2.7.10.1 (Anaplastic Lymphoma Kinase)
RN  - EC 2.7.10.1 (Receptor Protein-Tyrosine Kinases)
SB  - IM
MH  - Adult
MH  - Aged
MH  - Aged, 80 and over
MH  - Anaplastic Lymphoma Kinase
MH  - Antibodies
MH  - Crizotinib
MH  - Female
MH  - Humans
MH  - *Immunohistochemistry
MH  - *In Situ Hybridization, Fluorescence
MH  - Lung Neoplasms/drug therapy/*enzymology/*genetics
MH  - Male
MH  - Middle Aged
MH  - Protein Kinase Inhibitors/therapeutic use
MH  - Pyrazoles/therapeutic use
MH  - Pyridines/therapeutic use
MH  - ROC Curve
MH  - Receptor Protein-Tyrosine Kinases/analysis/*genetics
MH  - Translocation, Genetic
PMC - PMC4132045
EDAT- 2014/05/03 06:00
MHDA- 2015/01/31 06:00
PMCR- 2014/08/13
CRDT- 2014/05/03 06:00
PHST- 2014/05/03 06:00 [entrez]
PHST- 2014/05/03 06:00 [pubmed]
PHST- 2015/01/31 06:00 [medline]
PHST- 2014/08/13 00:00 [pmc-release]
AID - S1556-0864(15)30299-9 [pii]
AID - 10.1097/JTO.0000000000000157 [doi]
PST - ppublish
SO  - J Thorac Oncol. 2014 Jun;9(6):769-74. doi: 10.1097/JTO.0000000000000157.