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Rapid screening for chromosomal aneuploidies using array-MLPA

BACKGROUND: Chromosome abnormalities, especially trisomy of chromosome 21, 13, or 18 as well as sex chromosome aneuploidy, are a well-established cause of pregnancy loss. Cultured cell karyotype analysis and FISH have been considered reliable detectors of fetal abnormality. However, results are usua...

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Autores principales: Yan, Jing-Bin, Xu, Miao, Xiong, Can, Zhou, Da-Wen, Ren, Zhao-Rui, Huang, Ying, Mommersteeg, Monique, van Beuningen, Rinie, Wang, Ying-Tai, Liao, Shi-Xiu, Zeng, Fanyi, Wu, Ying, Zeng, Yi-Tao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3111339/
https://www.ncbi.nlm.nih.gov/pubmed/21575262
http://dx.doi.org/10.1186/1471-2350-12-68
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author Yan, Jing-Bin
Xu, Miao
Xiong, Can
Zhou, Da-Wen
Ren, Zhao-Rui
Huang, Ying
Mommersteeg, Monique
van Beuningen, Rinie
Wang, Ying-Tai
Liao, Shi-Xiu
Zeng, Fanyi
Wu, Ying
Zeng, Yi-Tao
author_facet Yan, Jing-Bin
Xu, Miao
Xiong, Can
Zhou, Da-Wen
Ren, Zhao-Rui
Huang, Ying
Mommersteeg, Monique
van Beuningen, Rinie
Wang, Ying-Tai
Liao, Shi-Xiu
Zeng, Fanyi
Wu, Ying
Zeng, Yi-Tao
author_sort Yan, Jing-Bin
collection PubMed
description BACKGROUND: Chromosome abnormalities, especially trisomy of chromosome 21, 13, or 18 as well as sex chromosome aneuploidy, are a well-established cause of pregnancy loss. Cultured cell karyotype analysis and FISH have been considered reliable detectors of fetal abnormality. However, results are usually not available for 3-4 days or more. Multiplex ligation-dependent probe amplification (MLPA) has emerged as an alternative rapid technique for detection of chromosome aneuploidies. However, conventional MLPA does not allow for relative quantification of more than 50 different target sequences in one reaction and does not detect mosaic trisomy. A multiplexed MLPA with more sensitive detection would be useful for fetal genetic screening. METHODS: We developed a method of array-based MLPA to rapidly screen for common aneuploidies. We designed 116 universal tag-probes covering chromosomes 13, 18, 21, X, and Y, and 8 control autosomal genes. We performed MLPA and hybridized the products on a 4-well flow-through microarray system. We determined chromosome copy numbers by analyzing the relative signals of the chromosome-specific probes. RESULTS: In a blind study of 161 peripheral blood and 12 amniotic fluid samples previously karyotyped, 169 of 173 (97.7%) including all the amniotic fluid samples were correctly identified by array-MLPA. Furthermore, we detected two chromosome X monosomy mosaic cases in which the mosaism rates estimated by array-MLPA were basically consistent with the results from karyotyping. Additionally, we identified five Y chromosome abnormalities in which G-banding could not distinguish their origins for four of the five cases. CONCLUSIONS: Our study demonstrates the successful application and strong potential of array-MLPA in clinical diagnosis and prenatal testing for rapid and sensitive chromosomal aneuploidy screening. Furthermore, we have developed a simple and rapid procedure for screening copy numbers on chromosomes 13, 18, 21, X, and Y using array-MLPA.
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spelling pubmed-31113392011-06-10 Rapid screening for chromosomal aneuploidies using array-MLPA Yan, Jing-Bin Xu, Miao Xiong, Can Zhou, Da-Wen Ren, Zhao-Rui Huang, Ying Mommersteeg, Monique van Beuningen, Rinie Wang, Ying-Tai Liao, Shi-Xiu Zeng, Fanyi Wu, Ying Zeng, Yi-Tao BMC Med Genet Technical Advance BACKGROUND: Chromosome abnormalities, especially trisomy of chromosome 21, 13, or 18 as well as sex chromosome aneuploidy, are a well-established cause of pregnancy loss. Cultured cell karyotype analysis and FISH have been considered reliable detectors of fetal abnormality. However, results are usually not available for 3-4 days or more. Multiplex ligation-dependent probe amplification (MLPA) has emerged as an alternative rapid technique for detection of chromosome aneuploidies. However, conventional MLPA does not allow for relative quantification of more than 50 different target sequences in one reaction and does not detect mosaic trisomy. A multiplexed MLPA with more sensitive detection would be useful for fetal genetic screening. METHODS: We developed a method of array-based MLPA to rapidly screen for common aneuploidies. We designed 116 universal tag-probes covering chromosomes 13, 18, 21, X, and Y, and 8 control autosomal genes. We performed MLPA and hybridized the products on a 4-well flow-through microarray system. We determined chromosome copy numbers by analyzing the relative signals of the chromosome-specific probes. RESULTS: In a blind study of 161 peripheral blood and 12 amniotic fluid samples previously karyotyped, 169 of 173 (97.7%) including all the amniotic fluid samples were correctly identified by array-MLPA. Furthermore, we detected two chromosome X monosomy mosaic cases in which the mosaism rates estimated by array-MLPA were basically consistent with the results from karyotyping. Additionally, we identified five Y chromosome abnormalities in which G-banding could not distinguish their origins for four of the five cases. CONCLUSIONS: Our study demonstrates the successful application and strong potential of array-MLPA in clinical diagnosis and prenatal testing for rapid and sensitive chromosomal aneuploidy screening. Furthermore, we have developed a simple and rapid procedure for screening copy numbers on chromosomes 13, 18, 21, X, and Y using array-MLPA. BioMed Central 2011-05-17 /pmc/articles/PMC3111339/ /pubmed/21575262 http://dx.doi.org/10.1186/1471-2350-12-68 Text en Copyright ©2011 Yan et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Technical Advance
Yan, Jing-Bin
Xu, Miao
Xiong, Can
Zhou, Da-Wen
Ren, Zhao-Rui
Huang, Ying
Mommersteeg, Monique
van Beuningen, Rinie
Wang, Ying-Tai
Liao, Shi-Xiu
Zeng, Fanyi
Wu, Ying
Zeng, Yi-Tao
Rapid screening for chromosomal aneuploidies using array-MLPA
title Rapid screening for chromosomal aneuploidies using array-MLPA
title_full Rapid screening for chromosomal aneuploidies using array-MLPA
title_fullStr Rapid screening for chromosomal aneuploidies using array-MLPA
title_full_unstemmed Rapid screening for chromosomal aneuploidies using array-MLPA
title_short Rapid screening for chromosomal aneuploidies using array-MLPA
title_sort rapid screening for chromosomal aneuploidies using array-mlpa
topic Technical Advance
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3111339/
https://www.ncbi.nlm.nih.gov/pubmed/21575262
http://dx.doi.org/10.1186/1471-2350-12-68
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