Abbott RealTime IDH2 is an in vitro polymerase chain reaction (PCR) assay for the qualitative detection of single nucleotide variants (SNVs) coding nine IDH2 R140 and R172 mutations (R140Q, R140L, R140G, R140W, R172K, R172M, R172G, R172S, and R172W) in DNA extracted from human bone marrow (EDTA collection tubes) or blood (EDTA collection tubes). 

Abbott RealTime IDH2 is for use with the Abbott m2000rt System.

Abbott RealTime IDH2 consists of two kits:

• Abbott RealTime IDH2 Amplification Reagent Kit (List No. 08N82‑090)
  That includes 9 vials sufficient for 24 tests:
  • Oligonucleotide Regent 1 to 4 (each vial 0.880 mL)
  • DNA Polymerase (4 vials – 0.051 mL/vial)
  • Activation Reagent (1 vial – 1.220 mL/vial)
• Abbott RealTime IDH2 Control Kit (List No. 08N82‑080)
  That includes 10 vials sufficient for 5 runs:
  • Negative Control (5 vials – 0.210 mL / vial)
  • Positive Control (5 vials – 0210 mL / vial).

Intended Use

Abbott RealTime IDH2 is an in vitro polymerase chain reaction (PCR) assay for the qualitative detection of single nucleotide variants (SNVs) coding nine IDH2 R140 and R172 mutations (R140Q, R140L, R140G, R140W, R172K, R172M, R172G, R172S, and R172W) in DNA extracted from human bone marrow (EDTA) or blood (EDTA). Abbott RealTime IDH2 is for use with the Abbott m2000rt System.

Indications For Use

Abbott RealTime IDH2 is indicated as an aid in identifying acute myeloid leukemia (AML) patients with an isocitrate dehydrogenase-2 (IDH2) mutation for treatment with IDHIFA^® (enasidenib).

ASSAY PROTOCOL

The Abbott RealTime IDH2 assay protocol includes the following steps:

        A. Manual preparation (ie, DNA extraction) of samples (specimens and controls) using the Abbott mSample Preparation SystemDNA (List No. 06K12 24) reagents to isolate and purify sample DNA.

        B. PCR assay setup using the sample eluates and the Abbott RealTime IDH2 Amplification Reagent Kit.

        C. Amplification/detection on the Abbott m2000rt instrument.

Specimens for Abbott RealTime IDH2 are processed manually using Abbott mSample Preparation SystemDNA (List No. 06K12 24) reagents to isolate and purify sample DNA. 

The Abbott RealTime IDH2 amplification reagents are combined into four amplification master mixes. 

The purified DNA sample is combined with the master mixes in an Abbott 96-Well Optical Reaction Plate, and the plate is transferred to the Abbott m2000rt instrument for amplification and detection of IDH2 mutations. 

The specimen result is automatically reported on the Abbott m2000rt workstation at run completion. Assay controls are included within each run and are processed through the DNA extraction, amplification, and detection steps of the assay to assess run validity.

Software parameters specific to Abbott RealTime IDH2 are contained in an assay application specification file, which is loaded onto the Abbott m2000rt instrument by using a CD ROM disk.

Refer to the WARNINGS AND PRECAUTIONS section of this package insert for instructions before preparing samples. 

LIMITATIONS OF THE ASSAY

• FOR IN VITRO DIAGNOSTIC USE
• Abbott RealTime IDH2 is for use with human blood (EDTA) and bone marrow aspirate (EDTA) specimens only.
• A “Not Detected” result does not preclude the presence of IDH2 mutations in the specimen. Assay results may be affected by inadequate specimen integrity, mutation content in the sample, and amount of amplifiable DNA.
• Abbott RealTime IDH2 is designed to detect IDH2 R140Q, R140L, R140G, R140W, R172K, R172M, R172G, R172S, and R172W mutations. Specimens with results reported as “Not Detected” may contain mutations that are not targeted by the assay.
• Analytical studies indicate that at high mutation percentage and high DNA input, the R140W probe can cross react with the R140G mutation. In this case, the software reports the mutation which is present at the highest level. All IDH2 mutations were correctly called in the analytical studies.
• Components contained within a kit are intended to be used together. Do not mix components from different kit lots. For example, do not use the IDH2 Oligonucleotide Reagents from Amplification kit lot X with the Enzyme Reagent from Amplification kit lot Y.