Kit comprising primers for amplifying alk kinase domain nucleic acids
AssigneeInsight Genetics, Inc.
InventorsDavid R. Hout, Rachel Skelton, John Handshoe, Kasey Lawrence, Brock Schweitzer
Life of this patent
Estimating the term…
Abstract
Disclosed are methods and kits for detecting the presence of a cancer in a subject and assessing the efficacy of treatments for the same. The disclosed method use reverse transcription polymerase chain reaction (RT-PCR) techniques to detect the presence of point mutations, truncations, or fusions of anaplastic lymphoma kinase.
Claims27
A kit for diagnosing a cancer comprising an anaplastic lymphoma kinase (ALK) fusion or oncogenic ALK transcript the kit comprising a forward and reverse primer pair that specifically hybridizes to an ALK kinase domain as set forth in SEQ ID NO:
- 1, and wherein at least one primer is a scorpion primer.
A method of diagnosing a cancer comprising an ALK fusion in a human subject the method comprising conducting a nucleic acid amplification process on a tissue sample from the subject and detecting the presence of nucleic acid associated with the ALK kinase domain in the tissue sample.
The method of claim 16, wherein the nucleic acid amplification process is reverse transcription polymerase chain reaction (RT-PCR) or real-time PCR.
The method of claim 17, wherein the RT-PCR or real-time PCR reaction comprises a forward and reverse primer pair that specifically hybridize to an ALK kinase domain sequence.
The method of claim 17 or 18 wherein the either the forward or reverse primer is a scorpion primer.
The method of claim 19, wherein the reverse primer comprises SEQ ID NO:
- 2.
A method of detecting the presence a cancer comprising an ALK fusion or overexpression of ALK in a tissue sample from a human subject with a cancer comprising performing a Polymerase Chain Reaction (PCR) reaction on the nucleic acid from the tissue sample, wherein the PCR reaction comprises at least one forward primer and at least one reverse primer that specifically hybridizes to a nucleic acid encoding the kinase domain of ALK as set forth in SEQ ID NO:
- 1, and detecting the presence or absence of the kinase domain of ALK, wherein the presence of the kinase domain of ALK indicates the presence of an ALK fusion.
The method of claim 23, wherein the PCR reaction is reverse transcription polymerase chain reaction (RT-PCR), real-time PCR, or real-time RT-PCR.
The method of claim 23 wherein the either the forward or reverse primer is a scorpion primer.
The method of claim 25, wherein the reverse primer comprises SEQ ID NO:
- 2.
A method of screening for an agent that inhibits a cancer comprising an ALK fusion comprising
- a) obtaining a tissue sample with a cancer comprising an ALK fusion;
- b) contacting the tissue sample with the agent
- c) extracting nucleic acid from the tissue sample;
- d) conducting a PCR, RT-PCR, real-time PCR, or real-time RT-PCR reaction on the mRNA from the tissue sample;
- wherein the reaction comprises a forward and reverse primer pair that specifically hybridizes to ALK kinase domain as set forth in SEQ ID NO:
- 1;
- and wherein a decrease in the amount of amplification product relative to an untreated control indicates an agent that can inhibit a cancer comprising an ALK fusion.
Dependencies are inferred from the claim text.