ALK Resistance Mutations and Cancer
Chromosomal translocations involving the anaplastic lymphoma kinase (ALK) gene have been found in various cancers, including 5-10% of non-small cell lung cancer (NSCLC). These in-frame rearrangements fuse the kinase domain of ALK with other genes, producing a chimeric protein with oncogenic potential.

Recently developed kinase inhibitors that target the kinase domain of ALK are currently used to treat ALK fusion positive NSCLC patients. However, the tumors stop responding to these drugs within months of the treatment. Several recent studies have identified secondary point mutations in the kinase domain of ALK that confer resistance to these kinase inhibitors. Thus, identification of these secondary ALK mutations in patients prior and during treatment with ALK inhibitors could enable clinicians to detect the development of resistance well in advance of detection of disease progression by imaging techniques (i.e. CT, MRI).

Assay Features
EntroGen’s ALK Resistance Mutation Analysis Kit is a real-time polymerase chain reaction (PCR)-based assay that uses mutant-specific primers and hydrolysis probes to identify the presence of ALK mutations in total RNA isolated from tumor tissues and bodily fluids. It contains specially formulated blend of enzymes and buffers that enable first strand cDNA synthesis and qualitative PCR for mutation detection in a single tube, in one step. Some of its features include:

• Fast and easy setup - one-step RT-PCR in a single tube
• Highly sensitive - detects mutations in a high range of RNA input (1pg - 2mg)
• Highly specific - uses hot start DNA polymerase to increase specificity
• Simple and accurate - Just 4 reactions per sample, includes internal control to ensure proper loading
• Universal - works on any real-time PCR instrument capable of detecting FAM™ and VIC®

Specifications
EntroGen’s ALK Resistance Mutation Analysis Kit detects the following ALK mutations: 

• L1152R (4362T>G)
• C1156Y (4374G>A)
• F1174L (4429 C>G)
• L1196M (4493C>A)

Equipment and Materials
The ALK Resistance Mutation Analysis Kit requires a real-time PCR instrument capable of detecting FAM and VIC fluorescent probes. This kit includes reagents required for the reverse PCR amplification/detection, as well as validated controls. Columns and reagents for RNA isolation are not included.

Intended Use
The ALK Resistance Mutation Analysis Kit is for Research Use Only (RUO). Not for use in diagnostic procedures.