[ Title ]
- Droplet Digital PCR for Absolute Quantification of EML4-ALK Gene
Rearrangement in Lung Adenocarcinoma
[ Journal ]
- JOURNAL OF MOLECULAR DIAGNOSTICS
[ Author ]
- Wang, QS
Yang, X
[ Abstract ]
- Crizotinib treatment significantly prolongs progression-free survival,
increases response rates, and improves the quality of Life in patients
with ALK-positive non small-cell lung cancer. Droplet Digital PCR
(ddPCR), a recently developed technique with high sensitivity and
specificity, was used in this study to evaluate the association between
the abundance of ALK rearrangements and crizotinib effectiveness. FFPE
tissues were obtained from 103 consecutive patients with Lung
adenocarcinoma. Fluorescent in situ hybridization (FISH) and ddPCR were
performed. The results revealed that 14 (13.6%) of the 103 patients were
positive by dual-color, break-apart FISH. Three variants (1, 2, and 3)
of the EML4-ALK gene rearrangements were detected. Thirteen of 14
ALK-positive cases identified by FISH were confirmed by ddPCR (four with
variant 1, two with variant 2, and seven with variant 3). The case
missed by ddPCR was identified as KIF5B-ALK gene rearrangement by
PCR-based direct sequencing. Sixteen patients were detected with low
copy numbers of EML4-ALK gene rearrangement, which failed to meet the
positive cutoff point of FISH. Two of them responded well to crizotinib
after unsuccessful chemotherapy. Our study indicates that ddPCR can be
used as a molecular analytical tool to accurately measure the EML4-ALK
rearrangement copy numbers in FFPE samples of lung adenocarcinoma
patients.
[ URL ]
- http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=Alerting&SrcApp=Alerting&DestApp=WOS&DestLinkType=FullRecord;UT=WOS:000360777700007