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Usefulness and Limitations of Multiple Ligation-Dependent Probe Amplification in Antithrombin Deficiency
Multiplex ligation-dependent probe amplification (MLPA) identifies genetic structural variants in SERPINC1 in 5% of cases with antithrombin deficiency (ATD), the most severe congenital thrombophilia. Our aim was to unravel the utility and limitations of MLPA in a large cohort of unrelated patients w...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10002544/ https://www.ncbi.nlm.nih.gov/pubmed/36902454 http://dx.doi.org/10.3390/ijms24055023 |
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author | Cifuentes, Rosa Padilla, José de la Morena-Barrio, María Eugenia de la Morena-Barrio, Belén Bravo-Pérez, Carlos Garrido-Rodríguez, Pedro Llamas, María Miñano, Antonia Vicente, Vicente Lozano, María Luisa Corral, Javier |
author_facet | Cifuentes, Rosa Padilla, José de la Morena-Barrio, María Eugenia de la Morena-Barrio, Belén Bravo-Pérez, Carlos Garrido-Rodríguez, Pedro Llamas, María Miñano, Antonia Vicente, Vicente Lozano, María Luisa Corral, Javier |
author_sort | Cifuentes, Rosa |
collection | PubMed |
description | Multiplex ligation-dependent probe amplification (MLPA) identifies genetic structural variants in SERPINC1 in 5% of cases with antithrombin deficiency (ATD), the most severe congenital thrombophilia. Our aim was to unravel the utility and limitations of MLPA in a large cohort of unrelated patients with ATD (N = 341). MLPA identified 22 structural variants (SVs) causing ATD (6.5%). MLPA did not detect SVs affecting introns (four cases), and the diagnosis was inaccurate in two cases according to long-range PCR or nanopore sequencing. MLPA was used to detect possible hidden SVs in 61 cases with type I deficiency with single nucleotide variations (SNVs) or small insertion/deletion (INDEL). One case had a false deletion of exon 7, as the 29-bp deletion affected an MLPA probe. We evaluated 32 variants affecting MLPA probes: 27 SNVs and 5 small INDELs. In three cases, MLPA gave false-positive results, all diagnosed as deletions of the affected exon: a small INDEL complex, and two SNVs affecting MLPA probes. Our study confirms the utility of MLPA to detect SVs in ATD, but also shows some limitations in detecting intronic SVs. MLPA renders imprecise and false-positive results for genetic defects which affect MLPA probes. Our results encourage the validation of MLPA results. |
format | Online Article Text |
id | pubmed-10002544 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-100025442023-03-11 Usefulness and Limitations of Multiple Ligation-Dependent Probe Amplification in Antithrombin Deficiency Cifuentes, Rosa Padilla, José de la Morena-Barrio, María Eugenia de la Morena-Barrio, Belén Bravo-Pérez, Carlos Garrido-Rodríguez, Pedro Llamas, María Miñano, Antonia Vicente, Vicente Lozano, María Luisa Corral, Javier Int J Mol Sci Article Multiplex ligation-dependent probe amplification (MLPA) identifies genetic structural variants in SERPINC1 in 5% of cases with antithrombin deficiency (ATD), the most severe congenital thrombophilia. Our aim was to unravel the utility and limitations of MLPA in a large cohort of unrelated patients with ATD (N = 341). MLPA identified 22 structural variants (SVs) causing ATD (6.5%). MLPA did not detect SVs affecting introns (four cases), and the diagnosis was inaccurate in two cases according to long-range PCR or nanopore sequencing. MLPA was used to detect possible hidden SVs in 61 cases with type I deficiency with single nucleotide variations (SNVs) or small insertion/deletion (INDEL). One case had a false deletion of exon 7, as the 29-bp deletion affected an MLPA probe. We evaluated 32 variants affecting MLPA probes: 27 SNVs and 5 small INDELs. In three cases, MLPA gave false-positive results, all diagnosed as deletions of the affected exon: a small INDEL complex, and two SNVs affecting MLPA probes. Our study confirms the utility of MLPA to detect SVs in ATD, but also shows some limitations in detecting intronic SVs. MLPA renders imprecise and false-positive results for genetic defects which affect MLPA probes. Our results encourage the validation of MLPA results. MDPI 2023-03-06 /pmc/articles/PMC10002544/ /pubmed/36902454 http://dx.doi.org/10.3390/ijms24055023 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Cifuentes, Rosa Padilla, José de la Morena-Barrio, María Eugenia de la Morena-Barrio, Belén Bravo-Pérez, Carlos Garrido-Rodríguez, Pedro Llamas, María Miñano, Antonia Vicente, Vicente Lozano, María Luisa Corral, Javier Usefulness and Limitations of Multiple Ligation-Dependent Probe Amplification in Antithrombin Deficiency |
title | Usefulness and Limitations of Multiple Ligation-Dependent Probe Amplification in Antithrombin Deficiency |
title_full | Usefulness and Limitations of Multiple Ligation-Dependent Probe Amplification in Antithrombin Deficiency |
title_fullStr | Usefulness and Limitations of Multiple Ligation-Dependent Probe Amplification in Antithrombin Deficiency |
title_full_unstemmed | Usefulness and Limitations of Multiple Ligation-Dependent Probe Amplification in Antithrombin Deficiency |
title_short | Usefulness and Limitations of Multiple Ligation-Dependent Probe Amplification in Antithrombin Deficiency |
title_sort | usefulness and limitations of multiple ligation-dependent probe amplification in antithrombin deficiency |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10002544/ https://www.ncbi.nlm.nih.gov/pubmed/36902454 http://dx.doi.org/10.3390/ijms24055023 |
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