Accuracy Of Genotype Imputation Research Articles

A fine-scale genetic map of the Japanese population.

Genetic maps are fundamental resources for linkage and association studies. A fine-scale genetic map can be constructed by inferring historical recombination events from the genome-wide structure of linkage disequilibrium-a non-random association of alleles among loci-by using population-scale sequencing data. We constructed a fine-scale genetic map and identified recombination hotspots from 10 092 551 bi-allelic high-quality autosomal markers segregating among 150 unrelated Japanese individuals whose genotypes were determined by high-coverage (30×) whole-genome sequencing, and the genotype quality was carefully controlled by using their parents' and offspring's genotypes. The pedigree information was also utilized for haplotype phasing. The resulting genome-wide recombination rate profiles were concordant with those of the worldwide population on a broad scale, and the resolution was much improved. We identified 9487 recombination hotspots and confirmed the enrichment of previously known motifs in the hotspots. Moreover, we demonstrated that the Japanese genetic map improved the haplotype phasing and genotype imputation accuracy for the Japanese population. The construction of a population-specific genetic map will help make genetics research more accurate.

A resampling-based approach to share reference panels

For many genome-wide association studies, imputing genotypes from a haplotype reference panel is a necessary step. Over the past 15 years, reference panels have become larger and more diverse, leading to improvements in imputation accuracy. However, the latest generation of reference panels is subject to restrictions on data sharing due to concerns about privacy, limiting their usefulness for genotype imputation. In this context, here we propose RESHAPE, a method that employs a recombination Poisson process on a reference panel to simulate the genomes of hypothetical descendants after multiple generations. This data transformation helps to protect against re-identification threats and preserves data attributes, such as linkage disequilibrium patterns and, to some degree, identity-by-descent sharing, allowing for genotype imputation. Our experiments on gold-standard datasets show that simulated descendants up to eight generations can serve as reference panels without substantially reducing genotype imputation accuracy.

Accuracy of genotype imputation of a low-density SNP array for the Amazon fish Colossoma macropomum.

In South America, Tambaqui (Colossoma macropomum) stands as the primary target for aquaculture, yet breeding programs for this Amazon native species are in their early stages. While high-density single nucleotide polymorphism (SNP) arrays are pivotal for aquaculture breeding, their costs can be prohibitive for non- or semi-industrial species. To overcome this, a cost-effective approach involves developing low-density SNP arrays followed by genotype imputation to higher densities. In this study, a 1K SNP array for tambaqui was created and validated, offering a balance between SNP quantity and genome representativity. The imputation accuracy from various SNP densities to a medium-density array was evaluated, with the 1K density demonstrating the best trade-off (accuracy of 0.93). This subset was further utilized to construct a commercial array through Agriseq™ targeted genotyping-by-sequencing, validated in 192 DNA samples, affirming its high quality for genotyping tambaqui. The low-density SNP array, with genome-wide coverage and high polymorphism, emerges as an effective tool for exploring genetic variation within diverse populations. Population analyses using the 1K panel proved to be an efficient tool for genetic characterization of sampled broodstocks, making it a valuable resource for genetic improvement programs targeting this Amazon native species.

Genotype imputation accuracy and the quality metrics of the minor ancestry in multi-ancestry reference panels.

Large-scale imputation reference panels are currently available and have contributed to efficient genome-wide association studies through genotype imputation. However, whether large-size multi-ancestry or small-size population-specific reference panels are the optimal choices for under-represented populations continues to be debated. We imputed genotypes of East Asian (180k Japanese) subjects using the Trans-Omics for Precision Medicine reference panel and found that the standard imputation quality metric (Rsq) overestimated dosage r2 (squared correlation between imputed dosage and true genotype) particularly in marginal-quality bins. Variance component analysis of Rsq revealed that the increased imputed-genotype certainty (dosages closer to 0, 1 or 2) caused upward bias, indicating some systemic bias in the imputation. Through systematic simulations using different template switching rates (θvalue) in the hidden Markov model, we revealed that the lower θ value increased the imputed-genotype certainty and Rsq; however, dosage r2 was insensitive to the θ value, thereby causing a deviation. In simulated reference panels with different sizes and ancestral diversities, the θ value estimates from Minimac decreased with the size of a single ancestry and increased with the ancestral diversity. Thus, Rsq could be deviated from dosage r2 for a subpopulation in the multi-ancestry panel, and the deviation represents different imputed-dosage distributions. Finally, despite the impact of the θ value, distant ancestries in the reference panel contributed only a few additional variants passing a predefined Rsq threshold. We conclude that the θ value substantially impacts the imputed dosage and the imputation quality metric value.

A diverse ancestrally-matched reference panel increases genotype imputation accuracy in a underrepresented population

Variant imputation, a common practice in genome-wide association studies, relies on reference panels to infer unobserved genotypes. Multiple public reference panels are currently available with variations in size, sequencing depth, and represented populations. Currently, limited data exist regarding the performance of public reference panels when used in an imputation of populations underrepresented in the reference panel. Here, we compare the performance of various public reference panels: 1000 Genomes Project, Haplotype Reference Consortium, GenomeAsia 100 K, and the recent Trans-Omics for Precision Medicine (TOPMed) program, when used in an imputation of samples from the Thai population. Genotype yields were assessed, and imputation accuracies were examined by comparison with high-depth whole genome sequencing data of the same sample. We found that imputation using the TOPMed panel yielded the largest number of variants (~ 271 million). Despite being the smallest in size, GenomeAsia 100 K achieved the best imputation accuracy with a median genotype concordance rate of 0.97. For rare variants, GenomeAsia 100 K also offered the best accuracy, although rare variants were less accurately imputable than common variants (30.3% reduction in concordance rates). The high accuracy observed when using GenomeAsia 100 K is likely attributable to the diverse representation of populations genetically similar to the study cohort emphasizing the benefits of sequencing populations classically underrepresented in human genomics.

Accurate rare variant phasing of whole-genome and whole-exome sequencing data in the UK Biobank

Phasing involves distinguishing the two parentally inherited copies of each chromosome into haplotypes. Here, we introduce SHAPEIT5, a new phasing method that quickly and accurately processes large sequencing datasets and applied it to UK Biobank (UKB) whole-genome and whole-exome sequencing data. We demonstrate that SHAPEIT5 phases rare variants with low switch error rates of below 5% for variants present in just 1 sample out of 100,000. Furthermore, we outline a method for phasing singletons, which, although less precise, constitutes an important step towards future developments. We then demonstrate that the use of UKB as a reference panel improves the accuracy of genotype imputation, which is even more pronounced when phased with SHAPEIT5 compared with other methods. Finally, we screen the UKB data for loss-of-function compound heterozygous events and identify 549 genes where both gene copies are knocked out. These genes complement current knowledge of gene essentiality in the human genome.

A Comprehensive Strategy Combining Feature Selection and Local Optimization Algorithm to Optimize the Design of Low-Density Chip for Genomic Selection

Design of low-density SNP chips provides an opportunity for wide application of genomic selection at lower cost. A novel strategy referred to as the “block-free” method is proposed in this study to select a subset of SNPs from a high-density chip to form a low-density panel. In this method, Feature Selection using a Feature Similarity (FSFS) algorithm was first performed to remove highly correlated SNPs, and then a Multiple-Objective, Local-Optimization (MOLO) algorithm was used to pick SNPs for the low-density panel. Two other commonly used methods called the “uniform” method and the “block-based” method were also implemented for comparison purposes. A real pig dataset with 7967 individuals from three breeds containing 43,832 SNPs was used for comparison of the methods. In terms of genotype imputation accuracy and genomic prediction accuracy, our strategy was superior in most cases when the densities were lower than 1K. The genotype imputation accuracy from the low-density chip compared to the original high-density chip was higher than 90% in all pig breeds as the density increased to 1K. In addition, the accuracies of predicted genomic breeding values (GEBV) calculated using the imputed panel were nearly 90% of estimates from the original chip for all traits and breeds. Our strategy is effective to design low-density chips by making full use of information of close relationships for genomic selection in animals and plants.

Ultra-low-coverage genome-wide association study—insights into gestational age using 17,844 embryo samples with preimplantation genetic testing

BackgroundVery low-coverage (0.1 to 1×) whole genome sequencing (WGS) has become a promising and affordable approach to discover genomic variants of human populations for genome-wide association study (GWAS). To support genetic screening using preimplantation genetic testing (PGT) in a large population, the sequencing coverage goes below 0.1× to an ultra-low level. However, the feasibility and effectiveness of ultra-low-coverage WGS (ulcWGS) for GWAS remains undetermined.MethodsWe built a pipeline to carry out analysis of ulcWGS data for GWAS. To examine its effectiveness, we benchmarked the accuracy of genotype imputation at the combination of different coverages below 0.1× and sample sizes from 2000 to 16,000, using 17,844 embryo PGT samples with approximately 0.04× average coverage and the standard Chinese sample HG005 with known genotypes. We then applied the imputed genotypes of 1744 transferred embryos who have gestational ages and complete follow-up records to GWAS.ResultsThe accuracy of genotype imputation under ultra-low coverage can be improved by increasing the sample size and applying a set of filters. From 1744 born embryos, we identified 11 genomic risk loci associated with gestational ages and 166 genes mapped to these loci according to positional, expression quantitative trait locus, and chromatin interaction strategies. Among these mapped genes, CRHBP, ICAM1, and OXTR were more frequently reported as preterm birth related. By joint analysis of gene expression data from previous studies, we constructed interrelationships of mainly CRHBP, ICAM1, PLAGL1, DNMT1, CNTLN, DKK1, and EGR2 with preterm birth, infant disease, and breast cancer.ConclusionsThis study not only demonstrates that ulcWGS could achieve relatively high accuracy of adequate genotype imputation and is capable of GWAS, but also provides insights into the associations between gestational age and genetic variations of the fetal embryos from Chinese population.

The NBDC-DDBJ imputation server facilitates the use of controlled access reference panel datasets in Japan

Accurate genotype imputation requires large-scale reference panel datasets. When conducting genotype imputation on the Japanese population, researchers can use such datasets under collaborative studies or controlled access conditions in public databases. We developed the NBDC-DDBJ imputation server, which securely provides users with a web user interface to execute genotype imputation on the server. Our benchmarking analysis showed that the accuracy of genotype imputation was improved by leveraging controlled access datasets to increase the number of haplotypes available for analysis compared to using publicly available reference panels such as the 1000 Genomes Project. The NBDC-DDBJ imputation server facilitates the use of controlled access datasets for accurate genotype imputation.

Aquaculture Molecular Breeding Platform (AMBP): a comprehensive web server for genotype imputation and genetic analysis in aquaculture.

It is of vital importance to understand the population structure, dissect the genetic bases of performance traits, and make proper strategies for selection in breeding programs. However, there is no single webserver covering the specific needs in aquaculture. We present Aquaculture Molecular Breeding Platform (AMBP), the first web server for genetic data analysis in aquatic species of farming interest. AMBP integrates the haplotype reference panels of 18 aquaculture species, which greatly improves the accuracy of genotype imputation. It also supports multiple tools to infer genetic structures, dissect the genetic architecture of performance traits, estimate breeding values, and predict optimum contribution. All the tools are coherently linked in a web-interface for users to generate interpretable results and evaluate statistical appropriateness. The webserver supports standard VCF and PLINK (PED, MAP) files, and implements automated pipelines for format transformation and visualization to simplify the process of analysis. As a demonstration, we applied the webserver to Pacific white shrimp and Atlantic salmon datasets. In summary, AMBP constitutes comprehensive resources and analytical tools for exploring genetic data and guiding practical breeding programs. AMBP is available at http://mgb.qnlm.ac.

Assessing accuracy of genotype imputation in the Afrikaner and Brahman cattle breeds of South Africa.

Imputation may be used to rescue genomic data from animals that would otherwise be eliminated due to a lower than desired call rate. The aim of this study was to compare the accuracy of genotype imputation for Afrikaner, Brahman, and Brangus cattle of South Africa using within- and multiple-breed reference populations. A total of 373, 309, and 101 Afrikaner, Brahman, and Brangus cattle, respectively, were genotyped using the GeneSeek Genomic Profiler 150K panel that contained 141,746 markers. Markers with MAF ≤ 0.02 and call rates ≤ 0.95 or that deviated from Hardy Weinberg Equilibrium frequency with a probability of ≤ 0.0001 were excluded from the data as were animals with a call rate ≤ 0.90. The remaining data included 99,086 SNPs and 360 Afrikaner, 75,291 SNPs and 288 animals Brahman, and 97,897 SNPs and 99 Brangus animals. A total of 7986, 7002, and 7000 SNP from 50 Afrikaner and Brahman and 30 Brangus cattle, respectively, were masked and then imputed using BEAGLE v3 and FImpute v2. The within-breed imputation yielded accuracies ranging from 89.9 to 96.6% for the three breeds. The multiple-breed imputation yielded corresponding accuracies from 69.21 to 88.35%. The results showed that population homogeneity and numerical representation for within and across breed strategies, respectively, are crucial components for improving imputation accuracies.

Accuracy of genotype imputation to whole genome sequencing level using different populations of Nile tilapia

A cost-effective strategy to obtain ultra-dense genomic information is to sequence part of population and perform imputation from lower density genotypes to sequence level for the remaining animals. The aims of this study were to evaluate the feasibility of genotype imputation from medium density to sequence level in Nile tilapia and to investigate the impacts of size and origin of reference population in the accuracy of imputation. Genomic DNA was extracted from fin-clip samples of 326 animals from 3 different populations (PA, PB and PC). After sequencing, alignment, variant calling and quality control of genotypes, approximately 4.6 million of single-nucleotide polymorphisms (SNPs) in common to all populations were retained and used for further imputation analyses. Four scenarios were evaluated to assess imputation accuracy on each population, including: two reference sizes (10 or 90% of animals of each reference population) and two reference origins (two different populations only or all three populations used as reference). The animals in the validation set had part of their genotypes masked keeping only 49,216 SNPs available and the accuracy of imputation was assessed using the correlation between the imputed and observed genotypes (R2). Imputation was carried out using FImpute3 software. At individual level, the R2 showed intermediate values ranging from 0.37 ± 0.04 to 0.56 ± 0.07 for PA, 0.43 ± 0.05 to 0.58 ± 0.08 for PB and 0.43 ± 0.05 to 0.58 ± 0.07 for PC. An increase in the R2 was observed when 90% of animals from the same population were used as reference in comparison to only 10% (0.37 ± 0.04 to 0.54 ± 0.07 for PA, 0.43 ± 0.05 to 0.57 ± 0.07 for PB and 0.43 ± 0.05 to 0.58 ± 0.07 for PC). At SNP level, the use of all three populations as reference yielded the best results in terms of number of SNPs imputed with accuracy greater than 0.8. On average, 676,233 ± 142,291, 666,559 ± 52,648 and 592,187 ± 89,663 SNPs were imputed with accuracy >0.8 for PA, PB and PC, respectively. Considering only these highly accurate imputed SNPs, the average imputation accuracy of samples was equal to 0.95 ± 0.06 for PA and 0.92 ± 0.07 for PB and PC, for scenarios that included more animals as reference (90% of same population as reference, two and three populations). There were no significant differences for R2 between scenarios that used 90% of animals from the same population and used animals from the three population as reference showing that the strategy of using information from other population to increase the reference population had minor effect on accuracy of imputation. In conclusion, it was feasible to impute from 50 K to approximately 700 K with high accuracy using tilapia sequence data. We also expect that the use of more animals from these populations or animals from ascending lines as reference could help in the imputation process to obtain millions of imputed SNPs with high accuracy.

Comparison of Genotype Imputation for SNP Array and Low-Coverage Whole-Genome Sequencing Data.

Genotype imputation is the term used to describe the process of inferring unobserved genotypes in a sample of individuals. It is a key step prior to a genome-wide association study (GWAS) or genomic prediction. The imputation accuracy will directly influence the results from subsequent analyses. In this simulation-based study, we investigate the accuracy of genotype imputation in relation to some factors characterizing SNP chip or low-coverage whole-genome sequencing (LCWGS) data. The factors included the imputation reference population size, the proportion of target markers /SNP density, the genetic relationship (distance) between the target population and the reference population, and the imputation method. Simulations of genotypes were based on coalescence theory accounting for the demographic history of pigs. A population of simulated founders diverged to produce four separate but related populations of descendants. The genomic data of 20,000 individuals were simulated for a 10-Mb chromosome fragment. Our results showed that the proportion of target markers or SNP density was the most critical factor affecting imputation accuracy under all imputation situations. Compared with Minimac4, Beagle5.1 reproduced higher-accuracy imputed data in most cases, more notably when imputing from the LCWGS data. Compared with SNP chip data, LCWGS provided more accurate genotype imputation. Our findings provided a relatively comprehensive insight into the accuracy of genotype imputation in a realistic population of domestic animals.

Towards a Cost-Effective Implementation of Genomic Prediction Based on Low Coverage Whole Genome Sequencing in Dezhou Donkey.

Low-coverage whole genome sequencing is a low-cost genotyping technology. Combined with genotype imputation approaches, it is likely to become a critical component of cost-effective genomic selection programs in agricultural livestock. Here, we used the low-coverage sequence data of 617 Dezhou donkeys to investigate the performance of genotype imputation for low-coverage whole genome sequence data and genomic prediction based on the imputed genotype data. The specific aims were as follows: 1) to measure the accuracy of genotype imputation under different sequencing depths, sample sizes, minor allele frequency (MAF), and imputation pipelines and 2) to assess the accuracy of genomic prediction under different marker densities derived from the imputed sequence data, different strategies for constructing the genomic relationship matrixes, and single-vs. multi-trait models. We found that a high imputation accuracy (>0.95) can be achieved for sequence data with a sequencing depth as low as 1x and the number of sequenced individuals ≥400. For genomic prediction, the best performance was obtained by using a marker density of 410K and a G matrix constructed using expected marker dosages. Multi-trait genomic best linear unbiased prediction (GBLUP) performed better than single-trait GBLUP. Our study demonstrates that low-coverage whole genome sequencing would be a cost-effective approach for genomic prediction in Dezhou donkey.

Accuracy of genotype imputation based on reference population size and marker density in Hanwoo cattle.

Recently, the cattle genome sequence has been completed, followed by developing a commercial single nucleotide polymorphism (SNP) chip panel in the animal genome industry. In order to increase statistical power for detecting quantitative trait locus (QTL), a number of animals should be genotyped. However, a high-density chip for many animals would be increasing the genotyping cost. Therefore, statistical inference of genotype imputation (low-density chip to high-density) will be useful in the animal industry. The purpose of this study is to investigate the effect of the reference population size and marker density on the imputation accuracy and to suggest the appropriate number of reference population sets for the imputation in Hanwoo cattle. A total of 3,821 Hanwoo cattle were divided into reference and validation populations. The reference sets consisted of 50k (38,916) marker data and different population sizes (500, 1,000, 1,500, 2,000, and 3,600). The validation sets consisted of four validation sets (Total 889) and the different marker density (5k [5,000], 10k [10,000], and 15k [15,000]). The accuracy of imputation was calculated by direct comparison of the true genotype and the imputed genotype. In conclusion, when the lowest marker density (5k) was used in the validation set, according to the reference population size, the imputation accuracy was 0.793 to 0.929. On the other hand, when the highest marker density (15k), according to the reference population size, the imputation accuracy was 0.904 to 0.967. Moreover, the reference population size should be more than 1,000 to obtain at least 88% imputation accuracy in Hanwoo cattle.

The impact of reference composition and genome build on the accuracy of genotype imputation in Australian Angus cattle

Context Genotype imputation is an effective method to increase the number of SNP markers available for an animal and thereby increase the overall power of genome-wide associations and accuracy of genomic predictions. It is also the key to achieve a common set of markers for all individuals when the original genotypes are obtained using multiple genotyping platforms. High accuracy of imputed genotypes is crucial to their utility. Aims In this study, we propose a method for the construction of a common set of medium density markers for imputation, which relies on keeping as much information as possible. We also investigated the impact of changing marker coordinates on the basis of the new bovine genome assembly, ARS-UCD 1.2, on imputation accuracy. Methods In total, 49 754 animals with 45 364 single nucleotide polymorphism markers were used in a 10-fold cross-validation to compare four different imputation scenarios. The four scenarios were based on two alternative designs for the reference datasets. (1) A traditional reference panel that was created using the overlapping SNP from five medium density arrays and (2) a composite reference panel created by combining SNPs across the five arrays. Each of the reference datasets was used to test imputation accuracy when the SNPs were aligned on the basis of two genome assemblies (UMD 3.1 and ARS-UCD 1.2). Key results Our results showed that a composite reference panel can achieve higher imputation accuracies than does a traditional overlap reference. Incorporating mapping information on the basis of the recent genome build slightly improved the imputation accuracies, especially for lower density chips. Conclusions Markers with unreliable mapping information and animals with low connectedness to the imputation reference dataset benefited the most from the ARS-UCD 1.2 assembly and composite reference respectively. Implications The presented method is straightforward and can be used to setup an optimal imputation for accurate inference of genotypes in Australian Angus cattle.

Genome-Wide Association Study and Cost-Efficient Genomic Predictions for Growth and Fillet Yield in Nile Tilapia (Oreochromis niloticus).

Fillet yield (FY) and harvest weight (HW) are economically important traits in Nile tilapia production. Genetic improvement of these traits, especially for FY, are lacking, due to the absence of efficient methods to measure the traits without sacrificing fish and the use of information from relatives to selection. However, genomic information could be used by genomic selection to improve traits that are difficult to measure directly in selection candidates, as in the case of FY. The objectives of this study were: (i) to perform genome-wide association studies (GWAS) to dissect the genetic architecture of FY and HW, (ii) to evaluate the accuracy of genotype imputation and (iii) to assess the accuracy of genomic selection using true and imputed low-density (LD) single nucleotide polymorphism (SNP) panels to determine a cost-effective strategy for practical implementation of genomic information in tilapia breeding programs. The data set consisted of 5,866 phenotyped animals and 1,238 genotyped animals (108 parents and 1,130 offspring) using a 50K SNP panel. The GWAS were performed using all genotyped and phenotyped animals. The genotyped imputation was performed from LD panels (LD0.5K, LD1K and LD3K) to high-density panel (HD), using information from parents and 20% of offspring in the reference set and the remaining 80% in the validation set. In addition, we tested the accuracy of genomic selection using true and imputed genotypes comparing the accuracy obtained from pedigree-based best linear unbiased prediction (PBLUP) and genomic predictions. The results from GWAS supports evidence of the polygenic nature of FY and HW. The accuracy of imputation ranged from 0.90 to 0.98 for LD0.5K and LD3K, respectively. The accuracy of genomic prediction outperformed the estimated breeding value from PBLUP. The use of imputation for genomic selection resulted in an increased relative accuracy independent of the trait and LD panel analyzed. The present results suggest that genotype imputation could be a cost-effective strategy for genomic selection in Nile tilapia breeding programs.

36 A large multi-breed reference panel and maximizes genotype imputation accuracy in cattle

Abstract The use of array-based SNP genotyping in the beef and dairy industries has produced an astounding amount of medium-to-low density genomic data in the last decade. While low-density assays work exceptionally well in the context of genomic prediction, they are less useful in mapping and causal variant discovery. This project focuses on maximizing imputation accuracies to the marker set of two high-density research assays, the Illumina Bovine HD, and the GGP-F250 which contains a large proportion of rare and potentially functional variants (~850,000 total SNPs). This 850K SNP set is well-suited for both imputation to sequence-level genotypes and direct downstream analysis. For testing, 310 animals from multiple breeds, all with observed HD and F250 genotypes, were downsampled to various commercial chip densities ranging from 8K–130K markers. We use both well-established and novel measures of imputation accuracy to categorize precisely where, why, and how imputation errors are made. These metrics provide insights into downstream interpretation and identify situations where caution should be exercised when analyzing imputed variants. We find that a large multi-breed composite imputation reference comprised of 36,131 samples with either HD and F250 genotypes significantly increases imputation accuracy compared to a standard within-breed reference panel, particularly at low minor allele frequencies. Breed composition information for each animal in our testing panel allowed us to identify how a breed’s representation in the reference panel affects the imputation accuracy of both purebred and admixed animals. Starting chip density also impacts imputation accuracy, but gains appear to plateau at around 50,000 markers. The addition of the F250’s rare variation to the reference panel increased the imputation accuracy of rare variants from the HD assay by an average of 4.32%. We expect this low MAF content from the F250 to have a similar positive impact on rare variant imputation at the sequence level. Early work using 850K imputed data in genomic predictions has shown substantial increases in both chip heritability and prediction accuracies. Using a large multi-breed reference and the best practices identified through this work will maximize imputation accuracies in virtually all cattle populations, particularly ones that are highly admixed with little or no available pedigree information.

Impacts of SNP genotyping call rate and SNP genotyping error rate on imputation accuracy inHolsteincattle

Single nucleotide polymorphism (SNP) chips have been widely used in genetic studies and breeding applications in animal and plant species. The quality of SNP genotypes is of paramount importance. More often than not, there are situations in which a number of genotypes may fail, requiring them to be imputed. There are also situations in which ungenotyped loci need to be imputed between different chips, or high-density genotypes need to be imputed based on low-density genotypes. Under these circumstances, the validity and reliability of subsequent data analyses is subject to the accuracy of these imputed genotypes. For justifying a better understanding of factors affecting imputation accuracy, in the present study, the impacts of SNP genotyping call rate and SNP genotyping error rate on the accuracy of genotype imputation were investigated under two scenarios in 20 116 U.S. Holstein cattle, each genotyped with a GGP 50K SNP chip. When the two factors were not correlated in scenario 1, simulated genotyping call rate varied from 50% to 100% and simulated genotyping error rate changed from 0% to 50%, with both factors being independent of each other. In scenario 2, genotyping error rates were correlated with genotyping call rate, and the relationship was set up by fitting a linear regression model between the two variables on a real dataset. That is, the simulated SNP call rate varied from 100% to 50% whereas the SNP genotyping rate changed from 0% to 13.55%. Finally, a 5-fold cross-validation was used to assess the subsequent imputation accuracy. The results showed that when original SNP genotyping call rate were independent of SNP genotyping error rate, the imputation accuracy did not change significantly with the original genotyping call rate (P>0.05), but it decreased significantly as the genotyping error rate increased (P<0.01). However, when original genotyping call rate was negatively correlated with genotyping error rate, the imputation error increased with elevated original genotyping error rate. In both scenarios, genotyping call rate needs to be no less than 0.90 in order to obtain 98% or higher genotype imputation accuracy. The present results can provide guidance for establishing quality assurance criteria for SNP genotyping in practice.

Examining the Impact of Imputation Errors on Fine-Mapping Using DNA Methylation QTL as a Model Trait.

Genetic variants disrupting DNA methylation at CpG dinucleotides (CpG-SNP) provide a set of known causal variants to serve as models to test fine-mapping methodology. We use 1716 CpG-SNPs to test three fine-mapping approaches (Bayesian imputation-based association mapping, Bayesian sparse linear mixed model, and the J-test), assessing the impact of imputation errors and the choice of reference panel by using both whole-genome sequence (WGS), and genotype array data on the same individuals (n = 1166). The choice of imputation reference panel had a strong effect on imputation accuracy, with the 1000 Genomes Project Phase 3 (1000G) reference panel (n = 2504 from 26 populations) giving a mean nonreference discordance rate between imputed and sequenced genotypes of 3.2% compared to 1.6% when using the Haplotype Reference Consortium (HRC) reference panel (n = 32,470 Europeans). These imputation errors had an impact on whether the CpG-SNP was included in the 95% credible set, with a difference of ∼23% and ∼7% between the WGS and the 1000G and HRC imputed datasets, respectively. All of the fine-mapping methods failed to reach the expected 95% coverage of the CpG-SNP. This is attributed to secondary cis genetic effects that are unable to be statistically separated from the CpG-SNP, and through a masking mechanism where the effect of the methylation disrupting allele at the CpG-SNP is hidden by the effect of a nearby SNP that has strong linkage disequilibrium with the CpG-SNP. The reduced accuracy in fine-mapping a known causal variant in a low-level biological trait with imputed genetic data has implications for the study of higher-order complex traits and disease.