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Updated: Apr 17, 2026
Oncogenic Gene Fusion Detection Using Anchored Multiplex Polymerase Chain Reaction Followed by Next Generation Sequencing
Published on: July 5, 2019
[Analysis of EML4-ALK gene fusion mutation in patients 
with non-small cell lung cancer]
Xuzhou Wang1, Weisheng Chen2, Yinghao Yu1
1Department of Pathology, Fuzhou General Hospital of NanJing Military Rejion, Fuzhou 350025, China.
Background:
Non-small cell lung cancer (NSCLC) is the main type of lung cancer, and the related locus mutation detection research has become a hot direction of molecular targeted therapy, studying on gene mutation status of echinodem microtubule associated protein like 4-Anaplastic lymphoma kinase (EML4-ALK) and epidermal growth factor receptor (EGFR), detecting the sensitivity of EML4-ALK gene fusion and gene mutation of EGFR.
Methods:
EML4-ALK gene fusion in 85 cases of paraffin embedded tumor tissue and adjacent lung tissue was detected with the application of immunohistochemistry (IHC), Scorpions amplification refractory mutation system (Scorpions ARMS) fluorescence quantitative PCR and fluorescence in situ hybridization (FISH) technology, and EGFR gene in 18, 19, 20 and 21 exon mutation status was detected with the application of ARMS method.
Results:
In 115 cases of NSCLC, IHC showed 32 cases with ALK (D5F3) expression, the expression rate was 27.8%; ARMS showed 27 cases with EML4-ALK fusion gene mutation, the mutation detection rate was 23.5%; 53 cases were detected with EGFR mutation, the mutation rate was 46%. While FISH showed 23 cases with EML4-ALK fusion gene mutation, the detection rate was 20%, slightly lower than the ARMS detection results, suggesting that ARMS more sensitive.
Conclusions:
The application of IHC, ARMS fluorescence quantitative PCR and FISH technology can make a rapid and accurate evaluation of EML4-ALK gene fusion.
Insights
This study highlights the effectiveness of molecular testing in non-small cell lung cancer (NSCLC). Amplification Refractory Mutation System (ARMS) fluorescence quantitative PCR is a sensitive method for detecting echinoderm microtubule-associated protein-like 4-anaplastic lymphoma kinase (EML4-ALK) gene fusions.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Non-small cell lung cancer (NSCLC) is a leading cause of cancer death.
- Molecular targeted therapy for NSCLC relies on detecting specific gene mutations.
- Key targets include echinoderm microtubule-associated protein-like 4-anaplastic lymphoma kinase (EML4-ALK) fusions and epidermal growth factor receptor (EGFR) mutations.
Purpose of the Study:
- To evaluate the sensitivity and accuracy of different molecular detection methods for EML4-ALK gene fusion and EGFR mutations in NSCLC.
- To compare the performance of immunohistochemistry (IHC), Scorpions Amplification Refractory Mutation System (ARMS) fluorescence quantitative PCR, and fluorescence in situ hybridization (FISH) for EML4-ALK detection.
Main Methods:
- EML4-ALK gene fusion was assessed using IHC, ARMS, and FISH in 85 NSCLC tissue samples.
- EGFR gene mutations in exons 18, 19, 20, and 21 were detected using the ARMS method.
- A total of 115 NSCLC cases were analyzed for gene alterations.
Main Results:
- IHC detected ALK (D5F3) expression in 27.8% of cases (32/115).
- ARMS identified EML4-ALK fusion gene mutations in 23.5% of cases (27/115).
- EGFR mutations were found in 46% of cases (53/115).
- FISH detected EML4-ALK fusion in 20% of cases (23/115), indicating ARMS is more sensitive.
Conclusions:
- IHC, ARMS fluorescence quantitative PCR, and FISH are valuable tools for rapid and accurate evaluation of EML4-ALK gene fusion in NSCLC.
- ARMS demonstrated higher sensitivity for detecting EML4-ALK fusion compared to FISH.
- Accurate detection of EML4-ALK and EGFR alterations is crucial for guiding targeted therapy in NSCLC.
