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author | greg |
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date | Tue, 10 Jan 2023 20:40:45 +0000 |
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#!/bin/env Rscript library(parallel) library(hash) library(stringr) library(grid) library(gridExtra) library(optparse) options(width = 180) printif = function(string = NULL, condition){ if (condition) { print(string) } } findPlasmids = function(plasmidPSLFile = NULL, plasmidDatabase = NULL, amrPSLFile = NULL, amrDatabase, noAMR = FALSE, incPSLFile = NULL, incDatabase, noInc = FALSE, outputDirectory = NA, overwrite = TRUE, maxTargetLength = 300000, minQueryLength = 500, makeCircos = FALSE, minQueryCoverage = 1/2, minTargetCoverage = 1/2, searchDepth = NULL, verbosity = 0) { ## Verify the arguments argumentsGood = TRUE if (minQueryCoverage < .1 || minQueryCoverage > 1) { argumentsGood = FALSE message(paste('Minimum query coverage', minQueryCoverage, 'is outside of the range 0.1 <= x <= 1')) } if (minTargetCoverage < 0.02 || minTargetCoverage > 1) { argumentsGood = FALSE message(paste('Minimum target coverage', minTargetCoverage, 'is outside of the range 0.1 <= x <= 1')) } if (!argumentsGood){ message('There is a problem with the arguments') return() } printif(paste('Finding plasmids in', plasmidPSLFile), verbosity > 0) ## Keep track of the total score in case we doing a grid search totalPlasmidScore = 0 ## Check for the existence of the output directory, remove if it exists if (file.exists(outputDirectory)) { printif(paste('Removing existing output directory', outputDirectory), verbosity > 1) unlink(outputDirectory, recursive = TRUE) } printif(paste('Making output directory', outputDirectory), verbosity > 1) dir.create(outputDirectory) outputPrefix = paste0(outputDirectory, "/plasmids") ## Read in and filter the plasmid hits plasmidHits = read.table(plasmidPSLFile, row.names = NULL, header = FALSE, sep = '\t', stringsAsFactors = FALSE, skip = 5) colnames(plasmidHits) = c('match', 'mismatch', 'rep_m', 'Ns', 'tgap_c', 'tgap_b', 'qgap_c', 'qgap_b', 'strand', 'target', 'tlength', 'tstart', 'tstop', 'query', 'qlength', 'qstart', 'qstop', 'blocks', 'block_sizes', 'tstarts', 'qstarts') printif(paste("Sequence-plasmid hits:", nrow(plasmidHits)), verbosity > 0) plasmidHits = plasmidHits[order(plasmidHits[,'target'], -plasmidHits[,'qlength']), ] ## Toss out any hits missing information plasmidHits = plasmidHits[complete.cases(plasmidHits),] ## Toss out very long plasmid sequences -- probably actually genome chunks labeled incorrectly veryLongHits = sum(plasmidHits[,'tlength'] >= maxTargetLength) printif(paste('Removing', veryLongHits, 'hits greater than', maxTargetLength), verbosity > 0) plasmidHits = plasmidHits[plasmidHits[,'tlength'] <= maxTargetLength, ] printif(paste("Sequence-plasmid hits after removing very long plasmids:", nrow(plasmidHits)), verbosity > 0) ## Toss out very short query sequences -- probably junk or repeats veryShortQuery = sum(plasmidHits[,'qlength'] >= minQueryLength) printif(paste('Removing', veryShortQuery, 'queries less than', minQueryLength), verbosity > 0) plasmidHits = plasmidHits[plasmidHits[,'qlength'] >= minQueryLength, ] printif(paste("Sequence-plasmid hits after removing very short queries:", nrow(plasmidHits)), verbosity > 0) ## Toss out sequece-plasmid pairs below the coverage cutoff sequenceMatches = aggregate(x = plasmidHits[,'match',drop = FALSE], by = list(plasmidHits[,'query'], plasmidHits[,'target']), FUN = sum) printif(head(sequenceMatches), verbosity > 1) printif(paste('Sequence-plasmid pair matches:', paste(dim(sequenceMatches), collapse = 'x')), verbosity > 1) sequenceLengths = aggregate(x = plasmidHits[,'qlength', drop = FALSE], by = list(plasmidHits[,'query'], plasmidHits[,'target']), FUN = max) printif(head(sequenceLengths), verbosity > 1) printif(paste('Sequence-plasmid pair lengths:', paste(dim(sequenceLengths), collapse = 'x')), verbosity > 1) matchingFractions = cbind(sequenceMatches[,c(1,2)], sequenceMatches[,3] / sequenceLengths[,3]) colnames(matchingFractions) = c('query', 'target', 'fraction') printif(head(matchingFractions), verbosity > 1) printif(paste('Sequence-plasmid pair fractions:', paste(dim(matchingFractions), collapse = 'x')), verbosity > 1) matchingFractions = matchingFractions[matchingFractions[,'fraction'] >= minQueryCoverage,] printif(head(matchingFractions), verbosity > 1) printif(paste('Passing sequence-plasmid pair fractions:', paste(dim(matchingFractions), collapse = 'x')), verbosity > 1) aboveMinCoverage = apply(matchingFractions, 1, function(i){paste0(i['query'], '|', i['target'])}) plasmidHits = plasmidHits[apply(plasmidHits, 1, function(i){paste0(i['query'], '|', i['target'])}) %in% aboveMinCoverage, ] printif(paste("Sequence-plasmid hits after removing low-coverage hits:", nrow(plasmidHits)), verbosity > 0) ## Toss out plasmid sequences below the coverage cutoff targetMatches = aggregate(x = plasmidHits[,'match',drop = FALSE], by = list(plasmidHits[,'target']), FUN = sum) printif(head(targetMatches), verbosity > 1) printif(paste('Plasmid matches:', paste(dim(targetMatches), collapse = 'x')), verbosity > 1) targetLengths = aggregate(x = plasmidHits[,'tlength', drop = FALSE], by = list(plasmidHits[,'target']), FUN = max) printif(head(targetLengths), verbosity > 1) printif(paste('Plasmid lengths:', paste(dim(targetLengths), collapse = 'x')), verbosity > 1) matchingFractions = cbind(targetMatches[,1], targetMatches[,2] / targetLengths[,2]) colnames(matchingFractions) = c('target', 'fraction') printif(head(matchingFractions), verbosity > 1) printif(paste('Plasmid fractions:', paste(dim(matchingFractions), collapse = 'x')), verbosity > 1) matchingFractions = matchingFractions[matchingFractions[,'fraction'] >= minTargetCoverage,] printif(head(matchingFractions), verbosity > 1) printif(paste('Passing plasmid fractions:', paste(dim(matchingFractions), collapse = 'x')), verbosity > 1) aboveMinCoverage = matchingFractions[, 'target'] plasmidHits = plasmidHits[plasmidHits[, 'target'] %in% aboveMinCoverage, ] printif(paste("Sequence-plasmid hits after removing low-coverage hits:", nrow(plasmidHits)), verbosity > 0) ## If we're out of sequece-plasmid hits, then stop here if (nrow(plasmidHits) == 0) { message(paste('Not hits found')) return } ## Find out how much of each query (contig) is covered by each target (plasmid). ## Query coverage is constant and does not change as we assign contigs to plasmids queryCoverage = hash() queryMismatches = hash() for (i in 1:nrow(plasmidHits)) { if (!(i %% 1000)) { printif(paste('Processing hit', i, '/', nrow(plasmidHits)), verbosity > 0) } query = plasmidHits[i,'query'] target = plasmidHits[i, 'target'] ## Represent each sequence-plasmid hit as a series of 0/1 vectors that if (!has.key(query, queryCoverage)) { queryCoverage[[query]] = hash() queryMismatches[[query]] = hash() } if (!has.key(target, queryCoverage[[query]])) { queryCoverage[[query]][[target]] = rep(0, times = plasmidHits[i, 'qlength']) queryMismatches[[query]][[target]] = 0 } blockSizes = as.numeric(unlist(strsplit(x = plasmidHits[i,'block_sizes'], ','))) qBlockStarts = as.numeric(unlist(strsplit(x = plasmidHits[i,'qstarts'], ','))) for (j in 1:length(blockSizes)) { queryCoverage[[query]][[target]][qBlockStarts[j]:(qBlockStarts[j]+blockSizes[j])] = 1 } queryMismatches[[query]][[target]] = queryMismatches[[query]][[target]] + plasmidHits[i,'mismatch'] } ## Pull the full plasmid names from the blast database because BLAT/minimap2 doesn't report them, just the ID's targetIDs = plasmidHits[,'target'] targetIDs = gsub("\\|$", "", targetIDs) targetIDs = gsub(".*(\\|.*)$", "\\1", targetIDs) noDotIDs = gsub("\\|", "", targetIDs) noDotIDs = gsub("(^H[^.]+).[0-9]+$", "\\1", noDotIDs) noDotIDs = cbind(noDotIDs) targetFile = paste0(outputDirectory, '/targets.tsv', sep = '') write.table(file = targetFile, x = noDotIDs, quote = FALSE, row.names = FALSE, col.names = FALSE) command = paste('blastdbcmd -db', plasmidDatabase, '-entry_batch', targetFile, '| grep ">"') targetNames = system(command, intern = TRUE) printif(paste('Found', length(targetNames), 'target names for', length(targetIDs), 'targets.'), verbosity > 0) targetNames = gsub('^>.*\\| ', '', targetNames) targetNames = gsub('^>[^ ]+', '', targetNames) plasmidHits = cbind(plasmidHits, targetIDs, targetNames) printif(paste('Named hits:', nrow(plasmidHits)), verbosity > 1) #Pull just the plasmids out of the larget set of hits, i.e, make sure it has the word 'plasmid' in the description. plasmidHits = plasmidHits[grep('plasmid|vector', plasmidHits[,'targetNames'], ignore.case = TRUE), ,drop = FALSE] plasmidHits = plasmidHits[!grepl('tig0000|unnamed', plasmidHits[,'targetNames'], ignore.case = TRUE), , drop = FALSE] printif(paste("Sequece-plasmid hits after screening by name:", paste(dim(plasmidHits), collapse = 'x')), verbosity > 1) ## Stop if there is nothing left if (is.null(plasmidHits)) { message('Not hits found') return() } if (nrow(plasmidHits) == 0) { message('Not hits found') return() } ## Clean up the plasmid names -- they look like crap by default. plasmidNames = plasmidHits[,'targetNames'] plasmidNames = gsub(', comp.*', '', plasmidNames) plasmidNames = gsub(', contig.*', '', plasmidNames) plasmidNames = gsub(', partial.*', '', plasmidNames) plasmidNames = gsub('strain ', '', plasmidNames) plasmidNames = gsub('^ *', '', plasmidNames) plasmidNames = sub('^(cl\\|)(.*?) ', '', plasmidNames) plasmidNames = sub('subsp. (.*?) ', '', plasmidNames) plasmidNames = sub('serovar (.*?) ', '', plasmidNames) plasmidNames = sub('strain ', '', plasmidNames) plasmidNames = sub('plasmid$', '', plasmidNames) plasmidHits[,'targetNames'] = plasmidNames ## Just take the best hit for each plasmid, hence the head, 1 in the agg plasmidNames = aggregate(plasmidHits, by = list(plasmidHits[,'query']), FUN = head, 1) plasmidNames = plasmidNames[, 'targetNames', drop = FALSE] ## Find the set of plasmid coverage hits for each itteration usedContigs = c() plasmidMismatches = c() ## Order hits by the plasmid ID and the query length plasmidHits = plasmidHits[order(plasmidHits[,'target'], -plasmidHits[,'qlength']), ] ## Iterate, finding plasmids until we run out of usable sequence-plasmid its plasmidNumber = 0 plasmidResults = c() while (1) { ## Keep track of how many plasmids we have gone over plasmidNumber = plasmidNumber + 1 printif(paste('Sequence-plasmid hits left:', nrow(plasmidHits)), verbosity > 1) contigToPlasmid = hash() plasmidToContig = hash() plasmidCoverage = hash() plasmidCoverageWithRepeats = hash() contigCoverage = hash() ##Find contig/plasmid plasmid/contig pairs if (is.null(plasmidHits)) { break } if (nrow(plasmidHits) == 0) { break } repLengths = c() ## Find the coverage of each plasmid in the possible set by the contigs for (i in 1:nrow(plasmidHits)) { query = plasmidHits[i,'query'] target = plasmidHits[i,'target'] matches = plasmidHits[i,'match'] mismatches = plasmidHits[i,'mismatch'] score = matches - mismatches queryLength = plasmidHits[i,'qlength'] blockSizes = as.numeric(unlist(strsplit(plasmidHits[i, 'block_sizes'], ','))) queryStarts = as.numeric(unlist(strsplit(plasmidHits[i, 'qstarts'], ','))) targetStarts = as.numeric(unlist(strsplit(plasmidHits[i, 'tstarts'], ','))) ## Skip matches which have less than 50% of the bases from the contig on the plasmid -- probably not a good match if ((sum(queryCoverage[[query]][[target]]) - queryMismatches[[query]][[target]]) / queryLength <= minQueryCoverage) { next } targetLength = plasmidHits[i, 'tlength']; targetStart = plasmidHits[i, 'tstart']; targetStop = plasmidHits[i, 'tstop'] ## Relate this contig to this plasmid if (!has.key(query, contigToPlasmid)) { contigToPlasmid[[query]] = hash() contigCoverage[[query]] = hash() } if (!has.key(target, contigToPlasmid[[query]])) { contigToPlasmid[[query]][[target]] = score contigCoverage[[query]][[target]] = rep(0, queryLength) } else { contigToPlasmid[[query]][[target]] = contigToPlasmid[[query]][[target]] + score } ## Keep track of target(plasmid) coverage by the contigs if (!has.key(target, plasmidCoverage)) { plasmidCoverage[[target]] = rep(0, targetLength) plasmidCoverageWithRepeats[[target]] = rep(0, targetLength) plasmidToContig[[target]] = hash() plasmidMismatches[target] = 0 } penalized = FALSE for (j in 1:length(blockSizes)) { ## Keep track of all contig alignments to this plasmid, even with repeats plasmidCoverageWithRepeats[[target]][targetStarts[j]:(targetStarts[j] + blockSizes[j])] = 1 ## Skip if this region of the query sequence has already been assigned to this plasmid if (sum(contigCoverage[[query]][[target]][queryStarts[j]:(queryStarts[j] + blockSizes[j])] == 0) <= 50) { printif(paste('Sequence', query, 'already used for', target, '. ', paste0(queryStarts[j], '-', queryStarts[j] + blockSizes[j])), verbosity > 2) next } if (!penalized) { ## Penalty for every gap, only penalize once per match ## TODO: Penalize for gap length, not just once per gap #plasmidMismatches[target] = plasmidMismatches[target] + (length(blockSizes) - 1) * 100 plasmidMismatches[target] = plasmidMismatches[target] + mismatches * 5 penalized = TRUE } plasmidCoverage[[target]][targetStarts[j]:(targetStarts[j] + blockSizes[j])][contigCoverage[[query]][[target]][queryStarts[j]:(queryStarts[j] + blockSizes[j])] == 0] = plasmidCoverage[[target]][targetStarts[j]:(targetStarts[j] + blockSizes[j])][contigCoverage[[query]][[target]][queryStarts[j]:(queryStarts[j] + blockSizes[j])] == 0] + 1 contigCoverage[[query]][[target]][queryStarts[j]:(queryStarts[j] + blockSizes[j])] = 1 } ## Relate this plasmid to this contig if (!has.key(query, plasmidToContig[[target]])) { plasmidToContig[[target]][[query]] = score } else { plasmidToContig[[target]][[query]] = plasmidToContig[[target]][[query]] + score } if (target == 'NZ_GG692894.1' && query == 'contig_3_0') { print('NZ_GG692894.1') print(sum(plasmidCoverage[[target]])) print(sum(contigCoverage[[query]][[target]])) } } ## Get the best set of plasmids out, i.e., the set with the most bases matching between the contig and plasmid plasmidScores = c() for (thisPlasmid in keys(plasmidCoverage)){ thisPlasmidScore = sum(plasmidCoverage[[thisPlasmid]]) plasmidScores = c(plasmidScores, thisPlasmidScore) names(plasmidScores)[length(plasmidScores)] = thisPlasmid } plasmidScores = sort(plasmidScores - plasmidMismatches[names(plasmidScores)], dec = TRUE) if (length(plasmidScores) > 0) { printif('Highest scoring plasmids', verbosity > 1) printif(head(cbind(plasmidScores), 20), verbosity > 1) } ## Stop searching for plasmids if nothing matches well or we're out of hits if (length(plasmidScores) == 0) { printif('Out of plasmids', verbosity > 0) break } else if (max(plasmidScores) < 500) { printif('Out of min-scoring plasmids', verbosity > 0) break } ## For each matching plasmid, ordered by total bases matching the assembly, find the set of corresponding contigs plasmidToUse = 1 if (!is.null(searchDepth) && plasmidNumber <= length(searchDepth)) { plasmidToUse = searchDepth[plasmidNumber] } if (plasmidToUse > length(plasmidScores)) { plasmidToUse = length(plasmidScores) } plasmid = names(plasmidScores)[plasmidToUse] print(paste('Plasmid picked', plasmid)) totalPlasmidScore = totalPlasmidScore + plasmidScores[plasmid] printif(paste("Pulling sequences for", plasmid), verbosity > 0) ## Find contigs what haven't already been given to another plasmid so we can assign them next round plasmidContigs = keys(plasmidToContig[[plasmid]]) unusedContigs = plasmidContigs[!(plasmidContigs %in% usedContigs)] ## If no unused contigs that also map to this plasmid then skip it if (length(unusedContigs) == 0) { printif(paste("No unused sequences for", plasmid), verbosity > 1) next } ## Keep just the rows for this plasmid and which haven't already been used by another plasmid plasmidRows = plasmidHits[plasmidHits[,'target'] == plasmid,] plasmidRows = plasmidRows[plasmidRows[,'query'] %in% unusedContigs,,drop = FALSE] plasmidName = plasmidRows[1, 'targetNames'] plasmidID = plasmidRows[1, 'targetIDs'] ## Get the plasmid length command = paste('blastdbcmd -db', plasmidDatabase, '-entry', plasmidID, '-outfmt "%l"') plasmidLength = system(command, intern = TRUE) plasmidLength = rep(plasmidLength, nrow(plasmidRows)) ## How many bases from the plasmid are uncovered? plasmidMissing = rep(sum(plasmidCoverageWithRepeats[[as.character(plasmidID)]] == 0), nrow(plasmidRows)) ## Keep track of all of the contigs included usedContigs = c(usedContigs, unusedContigs) ## How many matching bases for each query sequence? thisPlasmidQuerySizes = c() thisPlasmidMatches = c() for (contig in unusedContigs) { thisPlasmidQuerySizes = c(thisPlasmidQuerySizes, length(queryCoverage[[contig]][[plasmid]])) thisPlasmidMatches = c(thisPlasmidMatches, sum(queryCoverage[[contig]][[plasmid]])) } ## Add this plasmid's hits onto the growing list of sequence-plasmid hits thisPlasmidResults = cbind(unusedContigs, plasmidName, as.character(plasmidID), thisPlasmidQuerySizes, thisPlasmidMatches, plasmidLength, plasmidMissing) colnames(thisPlasmidResults) = c('query.name', 'plasmid.name', 'plasmid.accession', 'query.size', 'aligned.bases', 'plasmid.size', 'plasmid.missing') thisPlasmidResults = thisPlasmidResults[order(-thisPlasmidMatches), ] plasmidResults = rbind(plasmidResults, thisPlasmidResults) ## Remove the contigs added to this plasmid from the list of plasmid/contig BLAT hits plasmidHits = plasmidHits[!(plasmidHits[,'query'] %in% usedContigs),] plasmidHits = plasmidHits[!(plasmidHits[,'target'] == plasmid),] } rownames(plasmidResults) = plasmidResults[,1] ## Check for the presence of AMR genes in this file if (!noAMR) { amrBEDFile = paste0(outputDirectory, '/amrMapping.bed') command = paste('cat', amrPSLFile, '| awk -F \'\\t\' \'($3 >= 80) && ($4 / $14 >= .95){OFS = "\t"; print $2,($9 < $10 ? $9 : $10),($9 < $10 ? $10 : $9),$1,$3/100,($9 < $10 ? "+" : "-")}\'', '| sort -k 1,1 -k 2,2n >', amrBEDFile) printif(command, verbosity > 1) system(command) ## Find local overlapping regions amrMergedBEDFile = paste0(outputDirectory, '/amrMergedMapping.bed') command = paste('bedtools merge -d -30 -i', amrBEDFile, '>', amrMergedBEDFile) printif(command, verbosity > 1) system(command) ## Find the best AMR gene for each region amrFinalBEDFile = paste0(outputDirectory, '/amrFinal.bed') command = paste('bedtools intersect', '-a', amrBEDFile, '-b', amrMergedBEDFile, '-f .9 -F .9', '-wao', '| awk \'$7 != "."\'', '| awk \'{OFS="\t";locus=$7"\t"$8"\t"$9; if($5 > s[locus]){s[locus]=$5;b[locus] = $1"\t"$2"\t"$3"\t"$4"\t"$5"\t"$6}}END{for(i in b){print i,b[i]}}\'', '>', amrFinalBEDFile) printif(command, verbosity > 1) system(command) ## Read the AMR results in and add them to the plasmid contigs amrResults = read.table(file = amrFinalBEDFile, header = FALSE, row.names = NULL, stringsAsFactors = FALSE, quote = '') amrResults[,7] = gsub('(_.*$)|(.*\\|)', '', amrResults[,7]) amrResults = aggregate(amrResults[ , 7, drop = FALSE], by = list(amrResults[,1]), function(i){paste(i, collapse = ', ')}) rownames(amrResults) = amrResults[,1] amrResults = amrResults[ , 2, drop = FALSE] print(amrResults) plasmidResults = cbind(plasmidResults, rep('', nrow(plasmidResults))) colnames(plasmidResults)[ncol(plasmidResults)] = 'amr' plasmidResults[rownames(plasmidResults) %in% rownames(amrResults), 'amr'] = amrResults[rownames(plasmidResults)[rownames(plasmidResults) %in% rownames(amrResults)], 1] } ## Check for the presence of incompatibility groups in this file if (!noInc) { incBEDFile = paste0(outputDirectory, '/incMapping.bed') command = paste('cat', incPSLFile, '| awk -F \'\\t\' \'($3 >= 80) && ($4 / $14 >= .95){OFS = "\t"; print $2,($9 < $10 ? $9 : $10),($9 < $10 ? $10 : $9),$1,$3/100,($9 < $10 ? "+" : "-")}\'', '| sort -k 1,1 -k 2,2n >', incBEDFile) printif(command, verbosity > 1) system(command) ## Find local overlapping regions incMergedBEDFile = paste0(outputDirectory, '/incMergedMapping.bed') command = paste('bedtools merge -d -30 -i', incBEDFile, '>', incMergedBEDFile) printif(command, verbosity > 1) system(command) ## Find the best INC group for each region incFinalBEDFile = paste0(outputDirectory, '/incFinal.bed') command = paste('bedtools intersect', '-a', incBEDFile, '-b', incMergedBEDFile, '-f .9 -F .9', '-wao', '| awk \'$7 != "."\'', '| awk \'{OFS="\t";locus=$7"\t"$8"\t"$9; if($5 > s[locus]){s[locus]=$5;b[locus] = $1"\t"$2"\t"$3"\t"$4"\t"$5"\t"$6}}END{for(i in b){print i,b[i]}}\'', '>', incFinalBEDFile) printif(command, verbosity > 1) system(command) ## Read the inc group results in and add them to the plasmid contigs incResults = read.table(file = incFinalBEDFile, header = FALSE, row.names = NULL, stringsAsFactors = FALSE, quote = '') incResults[,7] = gsub('(_.*$)|(.*\\|)', '', incResults[,7]) incResults = aggregate(incResults[ , 7, drop = FALSE], by = list(incResults[,1]), function(i){paste(i, collapse = ', ')}) rownames(incResults) = incResults[,1] incResults = incResults[ , 2, drop = FALSE] print(incResults) plasmidResults = cbind(plasmidResults, rep('', nrow(plasmidResults))) colnames(plasmidResults)[ncol(plasmidResults)] = 'inc' plasmidResults[rownames(plasmidResults) %in% rownames(incResults), 'inc'] = incResults[rownames(plasmidResults)[rownames(plasmidResults) %in% rownames(incResults)], 1] } ## Write the plasmid results to file plasmidChunksFile = paste0(outputDirectory, '/plasmids.tsv') write.table(file = plasmidChunksFile, x = plasmidResults, quote = FALSE, sep = '\t', row.names = FALSE, col.names = TRUE) ## Dump a sequence file of potential plasmid contigs plasmidSequenceFile = paste0(outputPrefix, '.fna') system(paste0('echo "" >', plasmidSequenceFile)) for (contig in plasmidResults[,'query.name']) { command = paste('faidx', paste0('assembly.fasta'), contig, '>>', plasmidSequenceFile) print(command) #system(command) } ## Return the total score of this round, in case we are doing a search return(totalPlasmidScore) } pChunks = function(plasmidPSLFile = NULL, plasmidDatabase = NULL, amrPSLFile = NULL, amrDatabase, noAMR = FALSE, incPSLFile = NULL, incDatabase, noInc = FALSE, outputDirectory = NA, overwrite = TRUE, maxTargetLength = 300000, minQueryLength = 200, makeCircos = FALSE, minQueryCoverage = 1/2, minTargetCoverage = 1/2, searchDepth = c(1), threads = 1, verbosity = 2) { print(plasmidDatabase) ## Verify the arguments argumentsGood = TRUE if (!file.exists(plasmidPSLFile)) { argumentsGood = FALSE message(paste('Plasmid PSL file', plasmidPSLFile, 'not found')) } if (is.na(outputDirectory)) { argumentsGood = FALSE message('Output directory not given') } if (file.exists(outputDirectory) && !overwrite) { argumentsGood = FALSE message(paste('Output directory', outputDirectory, 'already exists. Add overwrite = TRUE')) } if (minQueryCoverage < .1 || minQueryCoverage > 1) { argumentsGood = FALSE message(paste('Minimum query coverage', minQueryCoverage, 'is outside of the range 0.1 <= x <= 1')) } if (minTargetCoverage < 0.02 || minTargetCoverage > 1) { argumentsGood = FALSE message(paste('Minimum target coverage', minTargetCoverage, 'is outside of the range 0.1 <= x <= 1')) } if (!argumentsGood){ message('There is a problem with the arguments') return() } ## Check for the existence of the output directory, remove if it exists if (file.exists(outputDirectory)) { printif(paste('Removing existing output directory', outputDirectory), verbosity > 1) unlink(outputDirectory, recursive = TRUE) } printif(paste('Making output directory', outputDirectory), verbosity > 1) dir.create(outputDirectory) ## Default to c(1) for the plasmid search depth searchDepths = lapply(searchDepth, function(i){seq(i, 1)}) searchDepths = as.matrix(expand.grid(searchDepths)) print(searchDepths) plasmidScores = mclapply(1:nrow(searchDepths), function(i) { findPlasmids(plasmidPSLFile = plasmidPSLFile, plasmidDatabase = plasmidDatabase, amrPSLFile = amrPSLFile, amrDatabase, noAMR = noAMR, incPSLFile = incPSLFile, incDatabase, noInc = noInc, outputDirectory = paste0(outputDirectory, '/plasmids_', paste(searchDepths[i,], collapse = '_')), overwrite = overwrite, maxTargetLength = 300000, minQueryLength = 200, makeCircos = makeCircos, minQueryCoverage = 1/2, minTargetCoverage = 1/2, searchDepth = searchDepths[i,], ## Search depth i verbosity = verbosity) }, mc.cores = threads) plasmidScores = unlist(plasmidScores) print(cbind(searchDepths, plasmidScores)) ## Pick out the best set, penalizing for not taking the first penalties = (unlist(apply(searchDepths, 1, sum)) - ncol(searchDepths)) * 2500 print(penalties) plasmidScores = plasmidScores - penalties print(cbind(searchDepths, plasmidScores)) bestScoring = which.max(plasmidScores) bestScoringDirectory = paste0(outputDirectory, '/plasmids_', paste(searchDepths[bestScoring,], collapse = '_')) ## Link to the best scoring files files = c('plasmids.tsv', 'amrFinal.bed') commands = paste('ln -s ', paste0('"', bestScoringDirectory, '/', files, '"'), paste0(outputDirectory, '/', files)) printif(commands, verbosity >= 2) lapply(commands, system) } optionList = list( make_option('--plasmid_psl', type = 'character', default = NULL, help = 'Plasmid PSL database-v-contig output', metavar = '<PSL_FILE>'), make_option('--plasmid_database', type = 'character', default = NULL, help = 'Plasmid database', metavar = '<PLASMID_FASTA>'), make_option('--amr_database', type = 'character', default = NULL, help = 'AMR database', metavar = '<AMR_FASTA>'), make_option('--amr_blast', type = 'character', default = NULL, help = 'AMR blast output', metavar = '<BLAST_6>'), make_option('--output', type = 'character', default = NULL, help = 'Output dir', metavar = '<OUTPUT_DIR>'), make_option('--threads', type = 'numeric', default = NULL, help = 'Output dir', metavar = '<OUTPUT_DIR>'), make_option('--no_amr', action = 'store_true', default = FALSE, help = 'Don\'t run AMR'), make_option('--no_inc', action = 'store_true', default = FALSE, help = 'Don\'t run incompatibility groups') ) optParser = OptionParser(option_list = optionList) opt = parse_args(optParser) print(opt) pChunks(plasmidPSLFile = opt$'plasmid_psl', plasmidDatabase = opt$'plasmid_database', amrPSLFile = opt$'amr_blast', amrDatabase = opt$'amr_database', outputDirectory = opt$output, threads = opt$threads, # searchDepth = c(1,1), searchDepth = c(5,5), noAMR = TRUE, noInc = TRUE, verbosity = 2)