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AmoyDx EML4-ALK fusion gene detection kit

Manufactured by Amoy Diagnostics
Sourced in China

The AmoyDx EML4-ALK fusion gene detection kit is a laboratory equipment product used for the detection of the EML4-ALK fusion gene. It functions as a tool for the identification and analysis of this specific genetic alteration.

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5 protocols using AmoyDx EML4-ALK fusion gene detection kit

All patient samples were tested for EGFR by using the amplification refractory mutation system(ARMS)assay with the AmoyDxEGFR 29 mutations detection kit and were tested forALK-status by using a reverse-transcriptase polymerase chain reaction (RT-PCR) assay with the AmoyDx EML4-ALK fusion gene detection kit (Amoy Diagnostics, Xiamen, People’s Republic of China). The amplification of β-actin was used to ensure the quality of RNA extracted. The ALK fusion variants screened by the AmoyDxEML4-ALK-fusion gene detection kit are shown in Appendix Table A1 (online only). AllALK fusion–positive orEGFR mutation–positive samples were validated by using direct sequencing. Details of the methodology have been described in previous studies.18 (link)–20 (link)
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Total RNA was extracted from tumor tissues to detect the EML4-ALK fusion by real-time polymerase chain reaction (qRT-PCR) using AmoyDx EML4-ALK Fusion Gene Detection Kit (Amoy Diagnostics, Xiamen, China), according to manufacturer’s instruction[3 (link)]. The amplified PCR product was subjected to direct sequencing, using AB3500xl DNA Sequencer (Applied Biosystems).
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The RNA samples were assayed using the AmoyDx® EML4-ALK Fusion Gene Detection Kit (Amoy Diagnostics, Xiamen, China) according to the manufacturer's instructions. Quantitative PCR was performed using Bioneer ExicyclerTM96 Real-Time Quantitative Thermal Block. The PCR condition was as recommended by the manufacturer. The EML4-ALK rearrangement status was determined by qRT-PCR with high-resolution melting (HRM) analysis.
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Three samples [1 histologic (Patient 1) and 2 cytologic (Patients 2 and 6)] were adequate to test for EML4-ALK fusion by qRT-PCR. In brief, total RNA was extracted from 3 slides of 4 μm thickness taken from the FFPE blocks using the AmoyDX RNA Isolation kit (Amoy Diagnostics, Xiamen, China), and mRNA was transcribed to cDNA at 42°C for 1 hour. EML4-ALK fusion was readily detected by qRT-PCR using the AmoyDx EML4-ALK Fusion Gene Detection kit (Amoy Diagnostics, Xiamen, China), according to the manufacturer's protocol.
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The RNA was extracted from FFPE tissues using a High Pure FFPE RNA Isolation Kit (Roche Diagnostics, Mannheim, Germany). EML4-ALK fusion transcripts were detected using the AmoyDx EML4-ALK fusion gene detection kit (Amoy Diagnostics, Xiamen, China). Fusion variants 1, 2, 3a, 3b, 4, 4′, 5a, 5b, 5′, and 8 were detected in 3 reactions according to the manufacturer’s protocol.
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