First‐time application of droplet digital PCR for methylation testing of the 11p15.5 imprinting regions
Abstract Background Beckwith‐Wiedemann syndrome and Silver‐Russel syndrome are two imprinting disorders caused by opposite molecular alterations in 11p15.5. With the current diagnostic tests, their molecular diagnosis is challenging due to molecular heterogeneity and mosaic occurrence of the most fr...
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| Format: | Article |
| Language: | English |
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Wiley
2023-12-01
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| Series: | Molecular Genetics & Genomic Medicine |
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| Online Access: | https://doi.org/10.1002/mgg3.2264 |
| _version_ | 1797389531276640256 |
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| author | Elia Schlaich Wouter H. G. Hubens Thomas Eggermann |
| author_facet | Elia Schlaich Wouter H. G. Hubens Thomas Eggermann |
| author_sort | Elia Schlaich |
| collection | DOAJ |
| description | Abstract Background Beckwith‐Wiedemann syndrome and Silver‐Russel syndrome are two imprinting disorders caused by opposite molecular alterations in 11p15.5. With the current diagnostic tests, their molecular diagnosis is challenging due to molecular heterogeneity and mosaic occurrence of the most frequent alterations. As the determination of precise (epi)genotype of patients is relevant as the basis for a personalized treatment, different approaches are needed to increase the sensitivity of diagnostic testing of imprinting disorders. Methods We established methylation‐specific droplet digital PCR approaches (MS‐ddPCR) for the two imprinting centers in 11p15.5, and analyzed patients with paternal uniparental disomy of chromosome 11p15.5 (upd(11)pat) and other imprinting defects in the region. The results were compared to those from MS‐MLPA (multiplex ligation‐dependent probe amplification) and MS‐pyrosequencing. Results MS‐ddPCR confirmed the molecular alterations in all patients and the results matched well with MS‐MLPA. The results of MS‐pyrosequencing varied between different runs, whereas MS‐ddPCR results were reproducible. Conclusion We show for the first time that MS‐ddPCR is a reliable and easy applicable method for determination of MS‐associated changes in imprinting disorders. It is therefore an additional tool for multimethod diagnostics of imprinting disorders suitable to improve the diagnostic yield. |
| first_indexed | 2024-03-08T22:57:21Z |
| format | Article |
| id | doaj.art-43a7dcfcf5934e0b8708de28a9b2f3b3 |
| institution | Directory Open Access Journal |
| issn | 2324-9269 |
| language | English |
| last_indexed | 2024-03-08T22:57:21Z |
| publishDate | 2023-12-01 |
| publisher | Wiley |
| record_format | Article |
| series | Molecular Genetics & Genomic Medicine |
| spelling | doaj.art-43a7dcfcf5934e0b8708de28a9b2f3b32023-12-16T05:49:02ZengWileyMolecular Genetics & Genomic Medicine2324-92692023-12-011112n/an/a10.1002/mgg3.2264First‐time application of droplet digital PCR for methylation testing of the 11p15.5 imprinting regionsElia Schlaich0Wouter H. G. Hubens1Thomas Eggermann2Institute for Human Genetics and Genome Medicine, Medical Faculty, RWTH Aachen University Aachen GermanyInstitute for Stem Cell Biology, Medical Faculty, RWTH Aachen University Aachen GermanyInstitute for Human Genetics and Genome Medicine, Medical Faculty, RWTH Aachen University Aachen GermanyAbstract Background Beckwith‐Wiedemann syndrome and Silver‐Russel syndrome are two imprinting disorders caused by opposite molecular alterations in 11p15.5. With the current diagnostic tests, their molecular diagnosis is challenging due to molecular heterogeneity and mosaic occurrence of the most frequent alterations. As the determination of precise (epi)genotype of patients is relevant as the basis for a personalized treatment, different approaches are needed to increase the sensitivity of diagnostic testing of imprinting disorders. Methods We established methylation‐specific droplet digital PCR approaches (MS‐ddPCR) for the two imprinting centers in 11p15.5, and analyzed patients with paternal uniparental disomy of chromosome 11p15.5 (upd(11)pat) and other imprinting defects in the region. The results were compared to those from MS‐MLPA (multiplex ligation‐dependent probe amplification) and MS‐pyrosequencing. Results MS‐ddPCR confirmed the molecular alterations in all patients and the results matched well with MS‐MLPA. The results of MS‐pyrosequencing varied between different runs, whereas MS‐ddPCR results were reproducible. Conclusion We show for the first time that MS‐ddPCR is a reliable and easy applicable method for determination of MS‐associated changes in imprinting disorders. It is therefore an additional tool for multimethod diagnostics of imprinting disorders suitable to improve the diagnostic yield.https://doi.org/10.1002/mgg3.2264imprinting disordersmosaicismMS‐ddPCRMS‐MLPAMS‐pyrosequencing |
| spellingShingle | Elia Schlaich Wouter H. G. Hubens Thomas Eggermann First‐time application of droplet digital PCR for methylation testing of the 11p15.5 imprinting regions Molecular Genetics & Genomic Medicine imprinting disorders mosaicism MS‐ddPCR MS‐MLPA MS‐pyrosequencing |
| title | First‐time application of droplet digital PCR for methylation testing of the 11p15.5 imprinting regions |
| title_full | First‐time application of droplet digital PCR for methylation testing of the 11p15.5 imprinting regions |
| title_fullStr | First‐time application of droplet digital PCR for methylation testing of the 11p15.5 imprinting regions |
| title_full_unstemmed | First‐time application of droplet digital PCR for methylation testing of the 11p15.5 imprinting regions |
| title_short | First‐time application of droplet digital PCR for methylation testing of the 11p15.5 imprinting regions |
| title_sort | first time application of droplet digital pcr for methylation testing of the 11p15 5 imprinting regions |
| topic | imprinting disorders mosaicism MS‐ddPCR MS‐MLPA MS‐pyrosequencing |
| url | https://doi.org/10.1002/mgg3.2264 |
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