New Quantitative Method to Identify NPM1 Mutations in Acute Myeloid Leukaemia

الملخص EN

Somatic mutations in the NPM1 gene, which encodes for nucleophosmin, have been reported to be the most frequent genetic abnormalities found in acute myeloid leukaemia (AML).

Their identification and quantification remain crucial for the patients’ residual disease monitoring.

We investigated a new method that could represent a novel reliable alternative to sequencing for its identification.

This method was based on high-resolution melting analysis in order to detect mutated patients and on an allele-specific oligonucleotide real-time quantitative polymerase chain reaction (ASO-RQ-PCR) for the identification and quantification of the transcripts carrying NPM1 mutations (NPM1m).

Few patients carrying known NPM1m enabled us to set up a table with the different primers’ ΔCT values, identifying a profile for each mutation type.

We then analysed a series of 337 AML patients' samples for NPM1 mutational status characterization and confirmed the ASO-RQ-PCR results by direct sequencing.

We identified some mutations in 86 samples, and the results were fully correlated in 100% of the 36 sequenced samples.

We also detected other rare NPM1m in two samples, that we confirmed by direct sequencing.

This highly specific method provides a novel quick, useful, and costless tool, easy to use in routine practice.