Highly Sensitive Detection of IDH2 Mutations in Acute Myeloid Leukemia

Petiti, Jessica; Rosso, Valentina; Croce, Eleonora; Franceschi, Vanessa; Andreani, Giacomo; Dragani, Matteo; Gobbi, Marco; Lunghi, Monia; Saglio, Giuseppe; Fava, Carmen; Cilloni, Daniela

doi:10.3390/jcm9010271

Cited by 12 publications

(8 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…On the other hand, dPCR has been considered as a complementary or even alternative method. In the hematological scenario, dPCR has already been employed for detection of hotspot mutations in many genes, such as JAK2 [19], IDH1/IDH2 [63,64], MYD88 [65], B-RAF [18,66], and c-KIT [67]. Each dPCR assay enables the detection of a given mutation by using specific primers and probes.…”

Section: Bcr-abl1 Mutation Testingmentioning

confidence: 99%

Molecular Testing in CML between Old and New Methods: Are We at a Turning Point?

Soverini

Bernardi

Galimberti

2020

JCM

View full text Add to dashboard Cite

Molecular monitoring of minimal residual disease (MRD) and BCR-ABL1 kinase domain (KD) mutation testing have a well consolidated role in the routine management of chronic myeloid leukemia (CML) patients, as they provide precious information for therapeutic decision-making. Molecular response levels are used to define whether a patient has an “optimal”, “warning”, or “failure” response to tyrosine kinase inhibitor (TKI) therapy. Mutation status may be useful to decide whether TKI therapy should be changed and which alternative TKI (or TKIs) are most likely to be effective. Real-time quantitative polymerase chain reaction (RQ-qPCR) and Sanger sequencing are currently the gold standard for molecular response monitoring and mutation testing, respectively. However, in recent years, novel technologies such as digital PCR (dPCR) and next-generation sequencing (NGS) have been evaluated. Here, we critically describe the main features of these old and novel technologies, provide an overview of the recently published studies assessing the potential clinical value of dPCR and NGS, and discuss how the state of the art might evolve in the next years.

show abstract

Section: Bcr-abl1 Mutation Testingmentioning

confidence: 99%

Molecular Testing in CML between Old and New Methods: Are We at a Turning Point?

Soverini

Bernardi

Galimberti

2020

JCM

View full text Add to dashboard Cite

show abstract

“…At diagnosis, ddPCR identified as mutated 16.2% of cases, PNA-PCR 14.8%, and Sanger 12.1%. This difference might be probably explained by the different sensitivity of the methods (1x10 −3 for ddPCR, 1% for PNA-PCR clamping, and 10–15% for Sanger) [ 15 ].…”

Section: Discussionmentioning

confidence: 99%

“…Nevertheless, the potential of using IDH mutations as markers for MRD is still a matter of debate; indeed, ELN guidelines do not suggest the use of IDH2 as a tool for MRD investigation, while some authors sustained that IDH2 might be a good biomarker [ 8 , 15 ]. Perhaps, the predictive/prognostic role of IDH2 mutations may be influenced either by the clinical context (for example target versus conventional therapies) or by some pathogenetic aspects, such as concomitant mutations that can represent an indirect sign of higher genomic instability or clonal branching [ 20 , 21 , 22 ].…”

Section: Discussionmentioning

confidence: 99%

Digital Droplet PCR is a Specific and Sensitive Tool for Detecting IDH2 Mutations in Acute Myeloid LeuKemia Patients

Grassi

Guerrini

Ciabatti

et al. 2020

Cancers

View full text Add to dashboard Cite

Isocitrate dehydrogenase 1 and 2 (IDH1 and IDH2) interfere with cellular metabolism contributing to oncogenesis. Mutations of IDH2 at R140 and R172 residues are observed in 20% of acute myeloid leukemias (AML), and the availability of the IDH2 inhibitor Enasidenib made IDH2 mutational screening a clinical need. The aim of this study was to set a new quantitative polymerase chain reaction (PCR) technique, the drop-off digital droplet PCR (drop-off ddPCR), as a sensitive and accurate tool for detecting IDH2 mutations. With this technique we tested 60 AML patients. Sanger sequencing identified 8/60 (13.5%) mutated cases, while ddPCR and the amplification refractory mutation system (ARMS) PCR, used as a reference technique, identified mutations in 13/60 (21.6%) cases. When the outcome of IDH2-mutated was compared to that of wild-type patients, no significant difference in terms of quality of response, overall survival, or progression-free survival was observed. Finally, we monitored IDH2 mutations during follow-up in nine cases, finding that IDH2 can be considered a valid marker of minimal residual disease (MRD) in 2/3 of our patients. In conclusion, a rapid screening of IDH2 mutations is now a clinical need well satisfied by ddPCR, but the role of IDH2 as a marker for MRD still remains a matter of debate.

show abstract

“…Furthermore, the PNA/DNA dimer cannot be amplified by DNA polymerase [ 17 ]. PNA-PCR clamping is based on the competition between a PNA probe and a primer to bind the same DNA sequence, and exploits the ability of PNA to hybridize to DNA and suppress amplification [ 18 , 19 , 20 , 21 , 22 ].…”

Section: Introductionmentioning

confidence: 99%

Detection of SF3B1 p.Lys700Glu Mutation by PNA-PCR Clamping in Myelodysplastic Syndromes and Myeloproliferative Neoplasms

Petiti

Itri

Signorino

et al. 2022

JCM

Self Cite

View full text Add to dashboard Cite

Mutations in SF3B1 are found in 20% of myelodysplastic syndromes and 5–10% of myeloproliferative neoplasms, where they are considered important for diagnosis and therapy decisions. Sanger sequencing and NGS are the currently available methods to identify SF3B1 mutations, but both are time-consuming and expensive techniques that are not practicable in most small-/medium-sized laboratories. To identify the most frequent SF3B1 mutation, p.Lys700Glu, we developed a novel fast and cheap assay based on PNA-PCR clamping. After setting the optimal PCR conditions, the limit of detection of PNA-PCR clamping was evaluated, and the method allowed up to 0.1% of mutated SF3B1 to be identified. Successively, PNA-PCR clamping and Sanger sequencing were used to blind test 90 DNA from patients affected by myelodysplastic syndromes and myeloproliferative neoplasms for the SF3B1 p.Lys700Glu mutation. PNA-PCR clamping and Sanger sequencing congruently identified 75 negative and 13 positive patients. Two patients identified as positive by PNA-PCR clamping were missed by Sanger analysis. The discordant samples were analyzed by NGS, which confirmed the PNA-PCR clamping result, indicating that these samples contained the SF3B1 p.Lys700Glu mutation. This approach could easily increase the characterization of myelodysplastic syndromes and myeloproliferative neoplasms in small-/medium-sized laboratories, and guide patients towards more appropriate therapy.

show abstract

Highly Sensitive Detection of IDH2 Mutations in Acute Myeloid Leukemia

Cited by 12 publications

References 29 publications

Molecular Testing in CML between Old and New Methods: Are We at a Turning Point?

Molecular Testing in CML between Old and New Methods: Are We at a Turning Point?

Digital Droplet PCR is a Specific and Sensitive Tool for Detecting IDH2 Mutations in Acute Myeloid LeuKemia Patients

Detection of SF3B1 p.Lys700Glu Mutation by PNA-PCR Clamping in Myelodysplastic Syndromes and Myeloproliferative Neoplasms

Contact Info

Product

Resources

About