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Open access
MDPI
Cancers
,  volume 17 ,  issue 5 ,  pages 854

Conventional PCR Versus Next Generation Sequencing for Diagnosis of FLT3, IDH and NPM1 Mutations in Acute Myeloid Leukemia: Results of the PETHEMA PCR-LMA Study

Blanca Boluda 1
, 
Rebeca Rodriguez-Veiga 1
, 
C. Sargas 2
, 
Rosa Ayala 3
, 
María José Larráyoz 4
, 
María Carmen Chillón 5
, 
Elena Soria‐Saldise 6
, 
Cristina Bilbao 7
, 
Esther Prados de la Torre 8
, 
Irene Navarro 1
, 
D. Martínez-Cuadrón 1, 9
, 
Cristina Gil 10
, 
T. Bernal 11
, 
Juan Bergua 12
, 
Lorenzo Algarra 13
, 
M J Tormo 14
, 
Pilar Martínez-Sanchez 3
, 
Estrella Carrillo-Cruz 6
, 
Josefina Serrano 8
, 
Juan Manuel Alonso Domínguez 15
, 
Raimundo García 16
, 
Maria Luz Amigo 17
, 
Pilar Herrera-Puente 18
, 
María José Sayas Lloris 19
, 
E. Lavilla 20
, 
María José García-Pérez 21
, 
Julia Morán 22
, 
Esther Pérez‐Santaolalla 23
, 
Natalia Alonso-Vence 24
, 
Ana Oliva 25
, 
Juan Antonio López 26
, 
Manuel Barrios 27
, 
María García Fortes 28
, 
Mayte Olave 29
, 
J. Labrador 30
, 
Joaquín Martínez-López 3
, 
María J Calasanz 4
, 
Ramón García-Sanz 5
, 
José Antonio Perez-Simon 6
, 
María T. Gómez-Casares 7
, 
Joaquín Sánchez-Garcia 8
, 
Yolanda Mendizabal 1
, 
Eva Barragán 2, 9
, 
Pau Montesinos 1, 9
1
 
Hematology Department, Hospital Universitari i Politécnic-IIS La Fe, 46026 Valencia, Spain | 
2
 
3
 
4
 
5
 
6
 
7
 
Hospital Universitario de Gran Canaria Dr. Negrín, 35010 Las Palmas de Gran Canaria, Spain | 
8
 
9
 
10
 
11
 
12
 
Hospital Universitario San Pedro de Alcántara, 10003 Cáceres, Spain | 
13
 
Hospital Universitario General de Albacete, 02008 Albacete, Spain | 
14
 
15
 
16
 
Hospital Universitari General de Castelló, 12004 Castellón, Spain | 
17
 
Hospital Universitario Morales Messeguer, 300008 Murcia, Spain | 
18
 
19
 
Hospital Universitari Dr. Peset, 46017 Valencia, Spain | 
20
 
21
 
Complejo Hospitalario Torrecardenas, 04009 Almería, Spain | 
22
 
23
 
Hospital Donostia, 20014 Donostia, Spain | 
24
 
Hospital Santiago de Compostela, 15706 Santiago, Spain | 
25
 
26
 
Hospital General Ciudad de Jaen, 23007 Jaen, Spain | 
27
 
28
 
Hospital U. Virgen de la Victoria, 29010 Málaga, Spain | 
29
 
Hospital Clínico U. Lozano Blesa, 50009 Zaragoza, Spain | 
30
 
Department of Hematology, Hospital University Burgos, 09006 Burgos, Spain | 
Publication type: Journal Article
Publication date: 2025-03-01
MDPI
scimago Q1
wos Q2
SJR: 1.462
CiteScore: 8.8
Impact factor: 4.4
ISSN: 20726694
Abstract

Background/Objectives: This PETHEMA PCR-LMA study aimed to evaluate whether mutations detected by NGS (VAF cut-off of ≥5%) correlate with NPM1, FLT3-ITD, FLT3-TKD, IDH1, and IDH2 mutations detected using conventional PCR (analytical sensitivity 3%) in a nationwide network of seven reference laboratories. Methods: Between 2019 and 2021, 1685 adult AML patients with at least one centralized sample (NGS or PCR) at primary diagnosis or relapse/refractory episode were included. Results: During this period, 1288 paired NGS/PCR samples (1094 at diagnosis, 103 at relapse and 88 at refractoriness) were analyzed. Considering PCR the gold-standard, for NPM1 NGS sensitivity was 98.5% and specificity 98.9%, for FLT3-ITD 73.8% and 99.6%, for FLT3-TKD 84.5% and 99.3%, for IDH1 98.7% and 98.7%, and for IDH2 99.1% and 97.7%, respectively. Overall concordance rate of positive results between NGS (and PCR was 95% (262/276) for NPM1, 72% (149/206) for FLT3-ITD, 74% (49/66) for FLT3-TKD, 87% (77/89) for IDH1 and 84% (107/127) for IDH2. Overall, median days from sample reception until report were 7 for PCR and 28 for NGS. Conclusions: This study shows high concordance between NPM1 and IDH results using PCR and NGS. However, sensible important discrepancies are observed for FLT3 mutations. In our context, rapid screening for these druggable mutations should be performed by conventional PCR.

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Boluda B. et al. Conventional PCR Versus Next Generation Sequencing for Diagnosis of FLT3, IDH and NPM1 Mutations in Acute Myeloid Leukemia: Results of the PETHEMA PCR-LMA Study // Cancers. 2025. Vol. 17. No. 5. p. 854.
GOST all authors (up to 50) Copy
Boluda B., Rodriguez-Veiga R., Sargas C., Ayala R., Larráyoz M. J., Chillón M. C., Soria‐Saldise E., Bilbao C., Prados de la Torre E., Navarro I., Martínez-Cuadrón D., Gil C., Bernal T., Bergua J., Algarra L., Tormo M. J., Martínez-Sanchez P., Carrillo-Cruz E., Serrano J., Alonso Domínguez J. M., García R., Amigo M. L., Herrera-Puente P., Sayas Lloris M. J., Lavilla E., García-Pérez M. J., Morán J., Pérez‐Santaolalla E., Alonso-Vence N., Oliva A., López J. A., Barrios M., García Fortes M., Olave M., Labrador J., Martínez-López J., Calasanz M. J., García-Sanz R., Perez-Simon J. A., Gómez-Casares M. T., Sánchez-Garcia J., Mendizabal Y., Barragán E., Montesinos P. Conventional PCR Versus Next Generation Sequencing for Diagnosis of FLT3, IDH and NPM1 Mutations in Acute Myeloid Leukemia: Results of the PETHEMA PCR-LMA Study // Cancers. 2025. Vol. 17. No. 5. p. 854.
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TY - JOUR
DO - 10.3390/cancers17050854
UR - https://www.mdpi.com/2072-6694/17/5/854
TI - Conventional PCR Versus Next Generation Sequencing for Diagnosis of FLT3, IDH and NPM1 Mutations in Acute Myeloid Leukemia: Results of the PETHEMA PCR-LMA Study
T2 - Cancers
AU - Boluda, Blanca
AU - Rodriguez-Veiga, Rebeca
AU - Sargas, C.
AU - Ayala, Rosa
AU - Larráyoz, María José
AU - Chillón, María Carmen
AU - Soria‐Saldise, Elena
AU - Bilbao, Cristina
AU - Prados de la Torre, Esther
AU - Navarro, Irene
AU - Martínez-Cuadrón, D.
AU - Gil, Cristina
AU - Bernal, T.
AU - Bergua, Juan
AU - Algarra, Lorenzo
AU - Tormo, M J
AU - Martínez-Sanchez, Pilar
AU - Carrillo-Cruz, Estrella
AU - Serrano, Josefina
AU - Alonso Domínguez, Juan Manuel
AU - García, Raimundo
AU - Amigo, Maria Luz
AU - Herrera-Puente, Pilar
AU - Sayas Lloris, María José
AU - Lavilla, E.
AU - García-Pérez, María José
AU - Morán, Julia
AU - Pérez‐Santaolalla, Esther
AU - Alonso-Vence, Natalia
AU - Oliva, Ana
AU - López, Juan Antonio
AU - Barrios, Manuel
AU - García Fortes, María
AU - Olave, Mayte
AU - Labrador, J.
AU - Martínez-López, Joaquín
AU - Calasanz, María J
AU - García-Sanz, Ramón
AU - Perez-Simon, José Antonio
AU - Gómez-Casares, María T.
AU - Sánchez-Garcia, Joaquín
AU - Mendizabal, Yolanda
AU - Barragán, Eva
AU - Montesinos, Pau
PY - 2025
DA - 2025/03/01
PB - MDPI
SP - 854
IS - 5
VL - 17
SN - 2072-6694
ER -
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@article{2025_Boluda,
author = {Blanca Boluda and Rebeca Rodriguez-Veiga and C. Sargas and Rosa Ayala and María José Larráyoz and María Carmen Chillón and Elena Soria‐Saldise and Cristina Bilbao and Esther Prados de la Torre and Irene Navarro and D. Martínez-Cuadrón and Cristina Gil and T. Bernal and Juan Bergua and Lorenzo Algarra and M J Tormo and Pilar Martínez-Sanchez and Estrella Carrillo-Cruz and Josefina Serrano and Juan Manuel Alonso Domínguez and Raimundo García and Maria Luz Amigo and Pilar Herrera-Puente and María José Sayas Lloris and E. Lavilla and María José García-Pérez and Julia Morán and Esther Pérez‐Santaolalla and Natalia Alonso-Vence and Ana Oliva and Juan Antonio López and Manuel Barrios and María García Fortes and Mayte Olave and J. Labrador and Joaquín Martínez-López and María J Calasanz and Ramón García-Sanz and José Antonio Perez-Simon and María T. Gómez-Casares and Joaquín Sánchez-Garcia and Yolanda Mendizabal and Eva Barragán and Pau Montesinos},
title = {Conventional PCR Versus Next Generation Sequencing for Diagnosis of FLT3, IDH and NPM1 Mutations in Acute Myeloid Leukemia: Results of the PETHEMA PCR-LMA Study},
journal = {Cancers},
year = {2025},
volume = {17},
publisher = {MDPI},
month = {mar},
url = {https://www.mdpi.com/2072-6694/17/5/854},
number = {5},
pages = {854},
doi = {10.3390/cancers17050854}
}
MLA
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MLA Copy
Boluda, Blanca, et al. “Conventional PCR Versus Next Generation Sequencing for Diagnosis of FLT3, IDH and NPM1 Mutations in Acute Myeloid Leukemia: Results of the PETHEMA PCR-LMA Study.” Cancers, vol. 17, no. 5, Mar. 2025, p. 854. https://www.mdpi.com/2072-6694/17/5/854.