PMID- 29251734
OWN - NLM
STAT- MEDLINE
DCOM- 20190205
LR  - 20230216
IS  - 1530-0307 (Electronic)
IS  - 0023-6837 (Linking)
VI  - 98
IP  - 3
DP  - 2018 Mar
TI  - Ligation-dependent RT-PCR: a new specific and low-cost technique to detect ALK, 
      ROS, and RET rearrangements in lung adenocarcinoma.
PG  - 371-379
LID - 10.1038/labinvest.2017.124 [doi]
AB  - Detection of anaplastic lymphoma kinase (ALK), ROS proto-oncogene 1 (ROS1), and 
      rearranged during transfection (RET) gene rearrangements in lung adenocarcinoma 
      is usually performed by immunohistochemistry (IHC) screening followed by 
      fluorescence in situ hybridization (FISH), which is an expensive and difficult 
      technique. Ligation-dependent reverse transcription polymerase chain reaction 
      (RT-PCR) multiplex technique can detect gene rearrangements using probes 
      specifically hybridized to either side of the break point. PCR products are then 
      sequenced by pyrosequencing or high throughput sequencing in order to identify 
      the two genes involved. The reagent cost is <15 dollars per patient and results 
      are available in 2 days. We have developed a 47-probe LD-RT-PCR kit especially 
      for lung adenocarcinomas. Thirty-nine lung adenocarcinomas were studied: 24 ALK+, 
      14 ROS1+, and 1 RET+. ALK+ and ROS1+ were IHC+ (D5F3 Ventana for ALK and D4D6 
      Cell Signaling Technology for ROS1) and all cases were FISH+ (Vysis ALK 
      Breakapart Probe Abbott for ALK, Zytolight SPEC ROS1 Dualcolor Breakapart Probe 
      for ROS1 and Zytolight SPEC RET Dual Color Breakapart for RET); 14 wild type 
      samples were included as negative controls. Using LD-RT-PCR, 15 rearrangements 
      (63%) were detected in the ALK cases (gene partner: EML4 in all cases), 9 
      rearrangements (64%) in the ROS1 cases (gene partners: CD74 in 8 cases and 
      SLC34A2 in 1 case) and 1 (100%) in the single RET case (gene partner: KIF5B). No 
      rearrangement was found in the 14 negative control cases. Negative cases using 
      LD-RT-PCR could be explained by the fact that some partner genes were not 
      included in our assay and therefore could not be detected. Because it is an 
      affordable, fast, and very simple technique, we propose using LD-RT-PCR when ALK 
      immunostaining is positive. For LD-RT-PCR-negative cases, samples should then be 
      analyzed by FISH.
FAU - Piton, Nicolas
AU  - Piton N
AD  - Rouen University Hospital, Department of Pathology, Rouen, France.
AD  - Normandie University, UNIROUEN, Inserm U1245, Rouen University Hospital, Rouen, 
      France.
FAU - Ruminy, Philippe
AU  - Ruminy P
AD  - Normandie University, UNIROUEN, Inserm U1245, Rouen University Hospital, Rouen, 
      France.
FAU - Gravet, Claire
AU  - Gravet C
AD  - Rouen University Hospital, Department of Pathology, Rouen, France.
FAU - Marchand, Vinciane
AU  - Marchand V
AD  - Normandie University, UNIROUEN, Inserm U1245, Rouen University Hospital, Rouen, 
      France.
FAU - Colasse, Elodie
AU  - Colasse E
AD  - Rouen University Hospital, Department of Pathology, Rouen, France.
FAU - Lamy, Aude
AU  - Lamy A
AD  - Rouen University Hospital, Department of Pathology, Rouen, France.
FAU - Naoures Mear, Cecile Le
AU  - Naoures Mear CL
AD  - Caen University Hospital, Department of Pathology, Caen, France.
FAU - Bibeau, Frederic
AU  - Bibeau F
AD  - Caen University Hospital, Department of Pathology, Caen, France.
FAU - Marguet, Florent
AU  - Marguet F
AD  - Rouen University Hospital, Department of Pathology, Rouen, France.
FAU - Guisier, Florian
AU  - Guisier F
AD  - Rouen University Hospital, Clinique Pneumologique, Rouen, France.
FAU - Salaun, Mathieu
AU  - Salaun M
AD  - Rouen University Hospital, Clinique Pneumologique, Rouen, France.
FAU - Thiberville, Luc
AU  - Thiberville L
AD  - Rouen University Hospital, Clinique Pneumologique, Rouen, France.
FAU - Jardin, Fabrice
AU  - Jardin F
AD  - Caen University Hospital, Department of Pathology, Caen, France.
FAU - Sabourin, Jean-Christophe
AU  - Sabourin JC
AD  - Rouen University Hospital, Department of Pathology, Rouen, France.
AD  - Normandie University, UNIROUEN, Inserm U1245, Rouen University Hospital, Rouen, 
      France.
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
DEP - 20171218
PL  - United States
TA  - Lab Invest
JT  - Laboratory investigation; a journal of technical methods and pathology
JID - 0376617
RN  - 0 (MAS1 protein, human)
RN  - 0 (Proto-Oncogene Mas)
RN  - 0 (Proto-Oncogene Proteins)
RN  - EC 2.7.10.1 (ALK protein, human)
RN  - EC 2.7.10.1 (Anaplastic Lymphoma Kinase)
RN  - EC 2.7.10.1 (Protein-Tyrosine Kinases)
RN  - EC 2.7.10.1 (Proto-Oncogene Proteins c-ret)
RN  - EC 2.7.10.1 (RET protein, human)
RN  - EC 2.7.10.1 (ROS1 protein, human)
SB  - IM
MH  - Adenocarcinoma/*genetics
MH  - Adult
MH  - Aged
MH  - Anaplastic Lymphoma Kinase/*genetics
MH  - Case-Control Studies
MH  - Female
MH  - Gene Rearrangement
MH  - Humans
MH  - Lung Neoplasms/*genetics
MH  - Male
MH  - Middle Aged
MH  - Protein-Tyrosine Kinases/*genetics
MH  - Proto-Oncogene Mas
MH  - Proto-Oncogene Proteins/*genetics
MH  - Proto-Oncogene Proteins c-ret/genetics
MH  - Reverse Transcriptase Polymerase Chain Reaction/*methods
MH  - Sensitivity and Specificity
EDAT- 2017/12/19 06:00
MHDA- 2019/02/06 06:00
CRDT- 2017/12/19 06:00
PHST- 2017/05/01 00:00 [received]
PHST- 2017/09/19 00:00 [revised]
PHST- 2017/09/21 00:00 [accepted]
PHST- 2017/12/19 06:00 [pubmed]
PHST- 2019/02/06 06:00 [medline]
PHST- 2017/12/19 06:00 [entrez]
AID - S0023-6837(22)01004-2 [pii]
AID - 10.1038/labinvest.2017.124 [doi]
PST - ppublish
SO  - Lab Invest. 2018 Mar;98(3):371-379. doi: 10.1038/labinvest.2017.124. Epub 2017 
      Dec 18.