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A protocol to impute haptoglobin alleles from 16q22.2 genotypes
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A protocol to impute haptoglobin haplotypes from surrounding genotypes computed from genotype array or whole genome sequence data using the reference panel from:

Boettger L., McCarroll S., et al. Recurring exon deletions in the HP (haptoglobin)
gene contribute to lower blood cholesterol levels. Nat. Genet. 48, 359–366 (2016)

This reference panel was generated as explained here. For any feedback, send an email to or


Install basic tools (Debian/Ubuntu specific):

sudo apt install wget gzip samtools bcftools plink1.9 openjdk-11-jre-headless

Preparation steps

mkdir -p $HOME/res

Download Beagle binary

wget -P $HOME/res/

Download reference panels

wget -P $HOME/res/{7,8}.vcf.gz

Run haptoglobin imputation from an input VCF

Run imputation using Beagle

build=38 # build=37
declare -A reg=( ["37"]="16:71070878-73097663" ["38"]="chr16:71036975-73063764" )

bcftools view --no-version "$vcf" -r ${reg[$build]} | \
  java -Xmx8g -jar $HOME/res/beagle.25Nov19.28d.jar gt=/dev/stdin \
  ref=$HOME/res/HP_Euroref_1kgOMNI_HM3_merged.GRCh$build.vcf.gz out="$out" \
  map=<(bcftools query -f "%CHROM\t%POS\n" $HOME/res/HP_Euroref_1kgOMNI_HM3_merged.GRCh$build.vcf.gz | \
  awk '{print $1"\t.\t"$2/1e7"\t"$2}')

Extract imputed haptoglobin alleles into a table

build=38 # build=37
declare -A reg=( ["37"]="16:72092044-72092044" ["38"]="chr16:72058145-72058145" )

bcftools index -ft "$out.vcf.gz" && \
bcftools query -f "[%SAMPLE\t%ALT\t%GT\n]" "$out.vcf.gz" -r ${reg[$build]} | tr -d '[<>]' | \
  awk -F"\t" -v OFS="\t" '{split($2,a,","); a["0"]="NA"; split($3,b,"|"); \
  print $1,a[b[1]],a[b[2]]}' > "$out.tsv"

Build the reference panels yourself

This section is only in case you want to build the reference panels yourself, you can skip it otherwise

Download GRCh37 human genome reference

wget -O- | \
  gzip -d > $HOME/res/human_g1k_v37.fasta
samtools faidx $HOME/res/human_g1k_v37.fasta

Download GRCh38 human genome reference

wget -O- | \
  gzip -d > $HOME/res/GCA_000001405.15_GRCh38_no_alt_analysis_set.fna
samtools faidx $HOME/res/GCA_000001405.15_GRCh38_no_alt_analysis_set.fna

Download haptoglobin reference panel in Beagle 3 format with marker positions for the GRCh37 human genome reference

wget -P $HOME/res/

Liftover marker positions for the GRCh38 human genome reference

chmod a+x liftOver
tail -n+2 $HOME/res/HP_Euroref_1kgOMNI_HM3_merged.bgl.phased | grep -v HP | \
    awk '{print "16\t"$2}' > $HOME/res/HP_Euroref_1kgOMNI_HM3_merged.GRCh37
tail -n+2 $HOME/res/HP_Euroref_1kgOMNI_HM3_merged.bgl.phased | grep -v HP | \
  awk '{print "chr16\t"$2-1"\t"$2}' | \
  ./liftOver /dev/stdin hg19ToHg38.over.chain.gz /dev/stdout /dev/stderr | \
  awk '{print $1"\t"$3}' > $HOME/res/HP_Euroref_1kgOMNI_HM3_merged.GRCh38

Generate haptoglobin reference panels in VCF format for both the GRCh37 and GRCh38 human genome references

declare -A fasta=( ["37"]="$HOME/res/human_g1k_v37.fasta" ["38"]="$HOME/res/GCA_000001405.15_GRCh38_no_alt_analysis_set.fna" )
declare -A chr=( ["37"]="16" ["38"]="chr16" )
declare -A pos=( ["37"]="72092044" ["38"]="72058145" )

grep HP $HOME/res/HP_Euroref_1kgOMNI_HM3_merged.bgl.phased | \
  awk 'BEGIN {x["AAAAABAAA"]=1; x["AABBBAAAB"]=4; x["BAABBAABA"]=3; x["BBAAAABAA"]=2}
  {for (i=3; i<=NF; i++) y[i]=y[i]$i}
  END {printf "<HP1A>,<HP1B>,<HP2A>,<HP2B>\t.\t.\t.\tGT";
  for (i in y) printf "\t"x[y[i]]}' | \
  sed 's/\t\([1-9]\)\t\([1-9]\)/\t\1|\2/g' > tmp

for build in 37 38; do
  tail -n+2 $HOME/res/HP_Euroref_1kgOMNI_HM3_merged.bgl.phased | grep -v HP | tr ' ' '\t' | cut -f3- | \
    paste $HOME/res/HP_Euroref_1kgOMNI_HM3_merged.GRCh$build - | \
    sed 's/\t\([ACGT]\)\t\([ACGT]\)/\t\1\2/g' | \
    bcftools convert --no-version --tsv2vcf /dev/stdin -c CHROM,POS,AA -f ${fasta[$build]} --samples \
    $(head -n1 $HOME/res/HP_Euroref_1kgOMNI_HM3_merged.bgl.phased | tr ' ' '\n' | \
    tail -n+3 | uniq | tr '\n' ',' | sed 's/,$//') | tr '/' '|' | \
    awk -v chr=${chr[$build]} -v pos=${pos[$build]} 'NR==FNR {line=$0}
    NR>FNR {if (line && $0!~"^#" && $2>pos) {print chr"\t"pos"\t.\tG\t"line; line=""} print}' tmp - | \
    bcftools view --no-version -Oz \
    -o $HOME/res/HP_Euroref_1kgOMNI_HM3_merged.GRCh$build.vcf.gz
/bin/rm tmp

Check for consistency of the haptoglobin reference panel

Convert the haptoglobin and 1000 Genomes project reference panels to plink and then merge to compute consistency

bcftools view --no-version $url -r 16:71070878-73097663 | \
  awk 'NF==2 {print "##contig=<ID=16,length=90354753>"} {print}' | \
  bcftools view --no-version -v snps | \
  bcftools annotate --no-version -x ID -I +'%CHROM:%POS:%REF:%ALT' | \
  $HOME/bin/plink --vcf /dev/stdin --keep-allele-order --const-fid --make-bed \
  --out ALL.chr16.integrated_phase1_v3.20101123.snps_indels_svs.genotypes

bcftools annotate --no-version -x ID -I +'%CHROM:%POS:%REF:%ALT' \
  $HOME/res/HP_Euroref_1kgOMNI_HM3_merged.GRCh37.vcf.gz | \
  $HOME/bin/plink --vcf /dev/stdin --biallelic-only --keep-allele-order --const-fid --make-bed \
  --out HP_Euroref_1kgOMNI_HM3_merged.GRCh37

plink --bfile HP_Euroref_1kgOMNI_HM3_merged.GRCh37 \
  --bmerge ALL.chr16.integrated_phase1_v3.20101123.snps_indels_svs.genotypes --merge-mode 6

You should get the following result

241696 overlapping calls, 241696 nonmissing in both filesets.
240611 concordant, for a concordance rate of 0.995511.
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