Long non‐coding RNAs: biomarkers for acute leukaemia subtypes |
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| Authors: | Carolina P. de Souza Melo Catharina B. Campos Juliana de Oliveira Rodrigues Joaquim C. Aguirre‐Neto Ângelo Atalla Mara A. D. Pianovski Edna K. Carbone Luciana B. Q. Lares Hélio Moraes‐Souza Shirlei Octacílio‐Silva Fabiano S. M. Pais Alessandro C. de Souza Ferreira Juliana G. Assumpção |
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| Affiliation: | 1. Laboratório BIOCOD Biotecnologia Ltda, Vespasiano, Brazil;2. Setor de Pesquisa e Desenvolvimento ‐ Instituto Hermes Pardini, Vespasiano, Brazil;3. Hospital Santa Casa de Misericórdia de Belo Horizonte, Belo Horizonte, Brazil;4. Hospital Universitário da Universidade Federal de Juiz de Fora, Juiz de Fora, Brazil;5. Hospital Erasto Gaertner, Curitiba, Brazil;6. Hospital Pequeno Príncipe, Curitiba, Brazil;7. Hospital S?o Jo?o de Deus, Divinópolis, Brazil;8. Hospital de Clínicas da Universidade Federal do Triangulo Mineiro, Uberlandia, Brazil;9. Departamento de Morfologia da Universidade Federal de Sergipe, Aracaju, Brazil;10. Grupo de Gen?mica e Biologia Computacional, Funda??o Oswaldo Cruz/Minas, Belo Horizonte, Brazil |
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| Abstract: | Accurate diagnosis of rare inherited anaemias is challenging, requiring a series of complex and expensive laboratory tests. Targeted next‐generation‐sequencing (NGS) has been used to investigate these disorders, but the selection of genes on individual panels has been narrow and the validation strategies used have fallen short of the standards required for clinical use. Clinical‐grade validation of negative results requires the test to distinguish between lack of adequate sequencing reads at the locations of known mutations and a real absence of mutations. To achieve a clinically‐reliable diagnostic test and minimize false‐negative results we developed an open‐source tool (CoverMi) to accurately determine base‐coverage and the ‘discoverability’ of known mutations for every sample. We validated our 33‐gene panel using Sanger sequencing and microarray. Our panel demonstrated 100% specificity and 99·7% sensitivity. We then analysed 57 clinical samples: molecular diagnoses were made in 22/57 (38·6%), corresponding to 32 mutations of which 16 were new. In all cases, accurate molecular diagnosis had a positive impact on clinical management. Using a validated NGS‐based platform for routine molecular diagnosis of previously undiagnosed congenital anaemias is feasible in a clinical diagnostic setting, improves precise diagnosis and enhances management and counselling of the patient and their family. |
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| Keywords: | lncRNA leukaemia biomarker gene expression profile classification model |
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