The Q-LAMP Method Represents a Valid and Rapid Alternative for the Detection of the BCR-ABL1 Rearrangement in Philadelphia-Positive Leukemias

Molecular detection of the fusion transcripts is necessary for the genetic confirmation of a chronic myeloid leukemia diagnosis and for the risk classification of acute lymphoblastic leukemia. mRNAs are usually identified using a conventional RT-PCR technique according to the BIOMED-1 method. In thi...

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Published in:	International journal of molecular sciences Vol. 20; no. 24; p. 6106
Main Authors:	Stella, Stefania, Gottardi, Enrico Marco, Favout, Valeria, Barragan Gonzalez, Eva, Errichiello, Santa, Vitale, Silvia Rita, Fava, Carmen, Luciano, Luigia, Stagno, Fabio, Grimaldi, Francesco, Pironi, Lucrezia, Sargas Simarro, Claudia, Vigneri, Paolo, Izzo, Barbara
Format:	Journal Article
Language:	English
Published:	Switzerland MDPI AG 04-12-2019 MDPI
Subjects:	Acute lymphoblastic leukemia Area Under Curve Assaying BCR protein Blood diseases Chromosomes Chronic myeloid leukemia Diagnosis Discrimination Fusion Proteins, bcr-abl - genetics Fusion Proteins, bcr-abl - metabolism Humans Isoforms Laboratories Leukemia Leukemia, Myelogenous, Chronic, BCR-ABL Positive - genetics Leukemia, Myelogenous, Chronic, BCR-ABL Positive - pathology Lymphatic leukemia Myeloid leukemia Nucleic Acid Amplification Techniques - methods Patients Philadelphia Chromosome Polymerase chain reaction Protein Isoforms - genetics Protein Isoforms - metabolism ROC Curve Thermal cycling chronic myeloid leukemia Q-LAMP rare transcripts e13a2 BCR-ABL1 e14a2
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Summary:	Molecular detection of the fusion transcripts is necessary for the genetic confirmation of a chronic myeloid leukemia diagnosis and for the risk classification of acute lymphoblastic leukemia. mRNAs are usually identified using a conventional RT-PCR technique according to the BIOMED-1 method. In this study, we evaluated 122 -positive samples with the Q-LAMP assay to establish if this technology may represent a valid alternative to the qualitative BIOMED-1 PCR technique usually employed for the detection and the discrimination of the common transcripts (p190 and p210 isoforms). We found a 100% concordance rate between the two methods. Specifically, the p190- and p210-positive samples were amplified by Q-LAMP with a median threshold time (Tt) of 26.70 min (range: 24.45-31.80 min) and 20.26 min (range: 15.25-34.57 min), respectively. A median time of 19.63 was observed in samples displaying both (e13a2/e14a2) p210 isoforms. Moreover, the Q-LAMP assay allowed recognition of the e13a2 and e14a2 isoforms (median Tts 18.48 for e13a2 vs. 26.08 min for e14a2; < 0.001). Finally, 20 samples harboring rare isoforms (e1a3, e13a3, e14a3, and e19a2) were correctly identified by the Q-LAMP assay. We conclude that the Q-LAMP assay may represent a faster and valid alternative to the qualitative BIOMED-1 RT-PCR for the diagnosis at -positive leukemias, especially when samples are analyzed in centers with restricted resources and/or limited technical expertise.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 These authors contributed equally to this work.
ISSN:	1422-0067 1661-6596 1422-0067
DOI:	10.3390/ijms20246106