Dataset Information

Detecting, quantifying, and discriminating the mechanism of mosaic chromosomal aneuploidies using MAD-seq.

ABSTRACT: Current approaches to detect and characterize mosaic chromosomal aneuploidy are limited by sensitivity, efficiency, cost, or the need to culture cells. We describe the mosaic aneuploidy detection by massively parallel sequencing (MAD-seq) capture assay and the MADSEQ analytical approach that allow low (<10%) levels of mosaicism for chromosomal aneuploidy or regional loss of heterozygosity to be detected, assigned to a meiotic or mitotic origin, and quantified as a proportion of the cells in the sample. We show results from a multi-ethnic MAD-seq (meMAD-seq) capture design that works equally well in populations of diverse racial and ethnic origins and how the MADSEQ analytical approach can be applied to exome or whole-genome sequencing data, revealing previously unrecognized aneuploidy or copy number neutral loss of heterozygosity in samples studied by the 1000 Genomes Project, cell lines from public repositories, and one of the Illumina Platinum Genomes samples. We have made the meMAD-seq capture design and MADSEQ analytical software open for unrestricted use, with the goal that they can be applied in clinical samples to allow new insights into the unrecognized prevalence of mosaic chromosomal aneuploidy in humans and its phenotypic associations.

SUBMITTER: Kong Y

PROVIDER: S-EPMC6028128 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature

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Detecting, quantifying, and discriminating the mechanism of mosaic chromosomal aneuploidies using MAD-seq.

Kong Yu Y Berko Esther R ER Marcketta Anthony A Maqbool Shahina B SB Simões-Pires Claudia A CA Kronn David F DF Ye Kenny Q KQ Suzuki Masako M Auton Adam A Greally John M JM

Genome research 20180517 7

Current approaches to detect and characterize mosaic chromosomal aneuploidy are limited by sensitivity, efficiency, cost, or the need to culture cells. We describe the mosaic aneuploidy detection by massively parallel sequencing (MAD-seq) capture assay and the <i>MADSEQ</i> analytical approach that allow low (<10%) levels of mosaicism for chromosomal aneuploidy or regional loss of heterozygosity to be detected, assigned to a meiotic or mitotic origin, and quantified as a proportion of the cells ...[more]

PMID: 29773658

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Project description:BackgroundChromosome 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.MethodsWe 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.ResultsIn 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.ConclusionsOur 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.

Project description:The presence of heterogeneity in susceptibility, differences between hosts in their likelihood of becoming infected, can fundamentally alter disease dynamics and public health responses, for example, by changing the final epidemic size, the duration of an epidemic, and even the vaccination threshold required to achieve herd immunity. Yet, heterogeneity in susceptibility is notoriously difficult to detect and measure, especially early in an epidemic. Here we develop a method that can be used to detect and estimate heterogeneity in susceptibility given contact by using contact tracing data, which are typically collected early in the course of an outbreak. This approach provides the capability, given sufficient data, to estimate and account for the effects of this heterogeneity before they become apparent during an epidemic. It additionally provides the capability to analyze the wealth of contact tracing data available for previous epidemics and estimate heterogeneity in susceptibility for disease systems in which it has never been estimated previously. The premise of our approach is that highly susceptible individuals become infected more often than less susceptible individuals, and so individuals not infected after appearing in contact networks should be less susceptible than average. This change in susceptibility can be detected and quantified when individuals show up in a second contact network after not being infected in the first. To develop our method, we simulated contact tracing data from artificial populations with known levels of heterogeneity in susceptibility according to underlying discrete or continuous distributions of susceptibilities. We analyzed these data to determine the parameter space under which we are able to detect heterogeneity and the accuracy with which we are able to estimate it. We found that our power to detect heterogeneity increases with larger sample sizes, greater heterogeneity, and intermediate fractions of contacts becoming infected in the discrete case or greater fractions of contacts becoming infected in the continuous case. We also found that we are able to reliably estimate heterogeneity and disease dynamics. Ultimately, this means that contact tracing data alone are sufficient to detect and quantify heterogeneity in susceptibility.

Dataset Information

Detecting, quantifying, and discriminating the mechanism of mosaic chromosomal aneuploidies using MAD-seq.

Publications

Detecting, quantifying, and discriminating the mechanism of mosaic chromosomal aneuploidies using MAD-seq.

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