Fluorescent and colorimetric RT-LAMP as a rapid and specific qualitative method for chronic myeloid leukemia diagnosis.

Autor: Marin AM; Laboratory for Applied Science and Technology in Health, Carlos Chagas Institute, Oswaldo Cruz Foundation (Fiocruz), Prof Algacyr Munhoz Mader 3775 Street, Curitiba, Paraná, ZIP 81350-010, Brazil., Zanette DL; Laboratory for Applied Science and Technology in Health, Carlos Chagas Institute, Oswaldo Cruz Foundation (Fiocruz), Prof Algacyr Munhoz Mader 3775 Street, Curitiba, Paraná, ZIP 81350-010, Brazil., Nardin JM; Erasto Gaertner Hospital, Dr. Ovande do Amaral 201 Street, Curitiba, Paraná, ZIP 81520-060, Brazil., Munhoz EC; Erasto Gaertner Hospital, Dr. Ovande do Amaral 201 Street, Curitiba, Paraná, ZIP 81520-060, Brazil., Blanes L; Laboratory for Applied Science and Technology in Health, Carlos Chagas Institute, Oswaldo Cruz Foundation (Fiocruz), Prof Algacyr Munhoz Mader 3775 Street, Curitiba, Paraná, ZIP 81350-010, Brazil., Soligo Sanchuki HB; Laboratory for Applied Science and Technology in Health, Carlos Chagas Institute, Oswaldo Cruz Foundation (Fiocruz), Prof Algacyr Munhoz Mader 3775 Street, Curitiba, Paraná, ZIP 81350-010, Brazil., Boçon de Araújo Munhoz F; Laboratory for Applied Science and Technology in Health, Carlos Chagas Institute, Oswaldo Cruz Foundation (Fiocruz), Prof Algacyr Munhoz Mader 3775 Street, Curitiba, Paraná, ZIP 81350-010, Brazil., de Oliveira Coelho B; Laboratory for Applied Science and Technology in Health, Carlos Chagas Institute, Oswaldo Cruz Foundation (Fiocruz), Prof Algacyr Munhoz Mader 3775 Street, Curitiba, Paraná, ZIP 81350-010, Brazil., Aoki MN; Laboratory for Applied Science and Technology in Health, Carlos Chagas Institute, Oswaldo Cruz Foundation (Fiocruz), Prof Algacyr Munhoz Mader 3775 Street, Curitiba, Paraná, ZIP 81350-010, Brazil. Electronic address: mateus.aoki@fiocruz.br.
Jazyk: angličtina
Zdroj: Analytical biochemistry [Anal Biochem] 2022 Mar 15; Vol. 641, pp. 114541. Date of Electronic Publication: 2021 Dec 29.
DOI: 10.1016/j.ab.2021.114541
Abstrakt: The detection of BCR-ABL1 mRNA transcripts is essential to molecular chronic myeloid leukemia (CML) diagnosis. In most cases, the RT-qPCR technique is performed as the gold standard diagnosis tool for clinical cases. However, this method requires expensive reagents and equipment, such as a real-time thermal cycler, probes and master mix. Consequently, the development and validation of simple and low-cost methods are essential for a rapid CML diagnosis in less specialized and equipped centers. In this study, we develop and demonstrate an accessible, rapid, and low-cost method using RT-LAMP for BCR-ABL1 detection in both cell lines and CML clinical samples, using fluorescent and colorimetric assays. Both methods demonstrated diagnostic specificity of 100% and while diagnostic sensitivity reaches more than 90% in samples with RT-qPCR cycle threshold above 31. The obtained data indicates that the proposed method here described is robust, specific and rapid approach for CML diagnosis with outstanding performance, especially for CML diagnostic procedure where present high BCR-ABL1 expression.
(Copyright © 2021. Published by Elsevier Inc.)
Databáze: MEDLINE
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