8cab21c8b8a5e84a3491b796bd6590c5d6905325 angie Thu Dec 4 15:45:54 2025 -0800 Added a couple samples to spare for reversion-riddled LF.7.6.5 diff --git src/hg/utils/otto/sarscov2phylo/updateLineageTreePb.sh src/hg/utils/otto/sarscov2phylo/updateLineageTreePb.sh index 9459bda28b0..9128b4b4a25 100755 --- src/hg/utils/otto/sarscov2phylo/updateLineageTreePb.sh +++ src/hg/utils/otto/sarscov2phylo/updateLineageTreePb.sh @@ -1,170 +1,173 @@ #!/bin/bash source ~/.bashrc set -beEu -x -o pipefail # Do not modify this script, modify the source tree copy: # kent/src/hg/utils/otto/sarscov2phylo/updateLineageTreePb.sh usage() { echo "usage: $0 buildDate [tree.pb.gz]" } if (( $# != 1 && $# != 2 )); then usage exit 1 fi buildDate=$1 if [ $# == 2 ]; then startingTree=$2 else startingTree=gisaidAndPublic.$buildDate.masked.pb.gz fi ottoDir=/hive/data/outside/otto/sarscov2phylo usherDir=~angie/github/usher matUtils=$usherDir/build/matUtils today=$(date +%F) cd $ottoDir/$buildDate # Remove sequences that have two or more reversions relative to their assigned clade/lineage. $matUtils summary -i $startingTree --node-stats node-stats -#*** Until BA.2.3.22 gets more samples that don't have bogus reversion on 25000 and 26577, -#*** exempt some samples; also, most of JP.1 has rev on 27383,27384: +# Until BA.2.3.22 gets more samples that don't have bogus reversion on 25000 and 26577, +# exempt some samples; also, most of JP.1 has rev on 27383,27384 and 2 of LF.7.6.5's 4 +# samples have revs. cat > pruneRevsExemptions <<EOF Germany/HH-RKI-I-1073418/2022|EPI_ISL_16330648|2022-12-18 Germany/SH-RKI-I-1071936/2022|EPI_ISL_16329344|2022-12-05 Germany/SH-RKI-I-1074649/2022|EPI_ISL_16396486|2022-12-12 Germany/SH-RKI-I-1071904/2022|EPI_ISL_16329314|2022-12-05 Germany/SH-RKI-I-1071924/2022|EPI_ISL_16329332|2022-12-05 Germany/SH-RKI-I-1071920/2022|EPI_ISL_16329328|2022-12-05 Germany/SH-RKI-I-1071899/2022|EPI_ISL_16329310|2022-12-05 Germany/HH-RKI-I-1029775/2022|EPI_ISL_15901009|2022-11-13 Germany/HH-RKI-I-1105132/2023|EPI_ISL_16940514|2023-01-27 OY160699.1 OY148358.1 OY125285.1 OY165180.1 OY176754.1 OY146516.1 OY142598.1 OY124642.1 OY143123.1 OY151190.1 SouthAfrica/NICD-N55605/2023|EPI_ISL_17859330|2023-05-22 SouthAfrica/NICD-N56015/2023|EPI_ISL_18125227|2023-06-15 SouthAfrica/NICD-N56013/2023|EPI_ISL_18125226|2023-06-15 SouthAfrica/NICD-R10184/2023|EPI_ISL_18341856|2023-09-05 SouthAfrica/NICD-R10169/2023|EPI_ISL_18341854|2023-09-05 SouthAfrica/NICD-R10455/2023|EPI_ISL_18341867|2023-09-12 +USA/NY-PRL-250108_82B06/2024|PQ898526.1|2024-12-22 +Egypt/MOH-CPHL-0003/2025|EPI_ISL_19975232|2025-04-13 EOF tawk '$5 > 1 {print $1;}' node-stats \ | grep -vFwf pruneRevsExemptions \ > pruneRevs $matUtils extract -i $startingTree \ -p -s pruneRevs -O -o gisaidAndPublic.$buildDate.masked.pruneRevs.pb.gz # Get node ID for root of lineage A, used as reference/root by Pangolin: $matUtils extract -i gisaidAndPublic.$buildDate.masked.pruneRevs.pb.gz -C clade-paths.prunedRevs lineageARoot=$(grep ^A$'\t' clade-paths.prunedRevs | cut -f 2) # Reroot protobuf to lineage A and restrict to low mutation density (highly supported nodes): $matUtils extract -i gisaidAndPublic.$buildDate.masked.pruneRevs.pb.gz \ --reroot $lineageARoot \ --max-mutation-density 2 \ -O -o gisaidAndPublic.$buildDate.masked.reroot.pb.gz # Reroot pango.clade-mutations.tsv grep -w ^A $scriptDir/pango.clade-mutations.tsv \ | sed -re 's/T28144C( > )?//; s/C8782T( > )?//;' \ > pango.clade-mutations.reroot.tsv grep -vw ^A $scriptDir/pango.clade-mutations.tsv \ | sed -re 's/\t/\tT8782C > C28144T > /;' \ | sed -re 's/T8782C >([ >ACGTN0-9,]+)C8782T/\1/; s/> +>/>/;' \ >> pango.clade-mutations.reroot.tsv # Mask additional bases at the beginning and end of the genome that pangolin masks after # aligning input sequences. set +x for ((i=56; $i <= 265; i++)); do echo -e "N${i}N" done > maskPangoEnds for ((i=29674; $i < 29804; i++)); do echo -e "N${i}N" done >> maskPangoEnds set -x $matUtils mask -i gisaidAndPublic.$buildDate.masked.reroot.pb.gz -c \ -m maskPangoEnds -o gisaidAndPublic.$buildDate.masked.reroot.pangoMasked.pb.gz # Preserve lineage annotations that survived rerooting and pango-masking $matUtils extract -i gisaidAndPublic.$buildDate.masked.reroot.pangoMasked.pb.gz \ -C clade-paths.reroot.pangoMasked tail -n+2 clade-paths.reroot.pangoMasked \ | grep '^[A-Za-z]' \ | cut -f 1,3 > lineageToPath.reroot.pangoMasked # Assign updated lineages on the rerooted & pango-masked tree, pango-only for pangolin: time $matUtils annotate -T 50 \ -i gisaidAndPublic.$buildDate.masked.reroot.pangoMasked.pb.gz \ -P lineageToPath.reroot.pangoMasked \ -M pango.clade-mutations.reroot.tsv \ -l \ -c lineageToName \ -f 0.95 \ -u mutations.pangoOnly \ -D details.pangoOnly \ -o gisaidAndPublic.$buildDate.masked.reroot.pangoOnly.pb.gz \ >& annotate.pangoOnly.out set +o pipefail grep 'Could not' annotate.pangoOnly.out | cat grep skip annotate.pangoOnly.out | cat set -o pipefail # Make a bunch of smaller trees and see how they do. mkdir -p /hive/users/angie/lineageTreeUpdate.$today cd /hive/users/angie/lineageTreeUpdate.$today for i in 0 1 2 3 4 5 6 7 8 9; do echo test.50.$i $matUtils extract -i $ottoDir/$buildDate/gisaidAndPublic.$buildDate.masked.reroot.pangoOnly.pb.gz \ -r 50 -o test.50.$i.pb $matUtils mask -i test.50.$i.pb -S -o test.50.$i.simp.pb done # Run pangolin on each subtree # 7.5-14.5m each job, ~3.5hrs total: conda activate pangolin for i in 0 1 2 3 4 5 6 7 8 9; do echo test.50.$i.simp time pangolin -t 80 --usher-tree test.50.$i.simp.pb --skip-scorpio \ --skip-designation-cache --no-temp --outdir subset_10000_0.pusher.test.50.$i.simp.out \ ../pangolin_eval/subset_10000_0.fa done # Summarize results for i in 0 1 2 3 4 5 6 7 8 9; do echo test.50.$i.simp tail -n+2 subset_10000_0.pusher.test.50.$i.simp.out/lineage_report.csv \ | awk -F, '{print $1 "\t" $2;}' \ | sort \ > subset_10000_0.pusher.test.50.$i.simp join -t$'\t' ../pangolin_eval/subset_10000_0.cogNameToLin subset_10000_0.pusher.test.50.$i.simp \ | tawk '$2 != $3' \ > subset_10000_0.pusher.test.50.$i.simp.diff cut -f 2,3 subset_10000_0.pusher.test.50.$i.simp.diff \ | sort | uniq -c | sort -nr > subset_10000_0.pusher.test.50.$i.simp.diffrank done wc -l subset_10000_0.*.diff | sort -n # Pick the subtree with the fewest differences set +o pipefail bestTree=$(wc -l subset_10000_0.*.diff | sort -n | head -1 \ | sed -re 's/.*pusher\.test\.50\.//; s/\.diff//') set -o pipefail echo $bestTree # Which lineages have the most differences? head subset_10000_0.pusher.test.50.$bestTree.diffrank