$LOAD_PATH.unshift(File.expand_path("../../", __FILE__)) require 'bio-blastxmlparser' require 'logger' module Quorum module SearchTools # # Blast Search Tool # class Blast class QuorumJob < ActiveRecord::Base self.table_name = "quorum_jobs" has_one :quorum_blastn_job, :foreign_key => "job_id" has_many :quorum_blastn_job_reports, :foreign_key => "blastn_job_id" has_one :quorum_blastx_job, :foreign_key => "job_id" has_many :quorum_blastx_job_reports, :foreign_key => "blastx_job_id" has_one :quorum_tblastn_job, :foreign_key => "job_id" has_many :quorum_tblastn_job_reports, :foreign_key => "tblastn_job_id" has_one :quorum_blastp_job, :foreign_key => "job_id" has_many :quorum_blastp_job_reports, :foreign_key => "blastp_job_id" end class QuorumBlastnJob < ActiveRecord::Base self.table_name = "quorum_blastn_jobs" belongs_to :quorum_job has_many :quorum_blastn_job_reports end class QuorumBlastnJobReport < ActiveRecord::Base self.table_name = "quorum_blastn_job_reports" belongs_to :quorum_blastn_job end class QuorumBlastxJob < ActiveRecord::Base self.table_name = "quorum_blastx_jobs" belongs_to :quorum_job has_many :quorum_blastx_job_reports end class QuorumBlastxJobReport < ActiveRecord::Base self.table_name = "quorum_blastx_job_reports" belongs_to :quorum_blastx_job end class QuorumTblastnJob < ActiveRecord::Base self.table_name = "quorum_tblastn_jobs" belongs_to :quorum_job has_many :quorum_tblastn_job_reports end class QuorumTblastnJobReport < ActiveRecord::Base self.table_name = "quorum_tblastn_job_reports" belongs_to :quorum_tblastn_job end class QuorumBlastpJob < ActiveRecord::Base self.table_name = "quorum_blastp_jobs" belongs_to :quorum_job has_many :quorum_blastp_job_reports end class QuorumBlastpJobReport < ActiveRecord::Base self.table_name = "quorum_blastp_job_reports" belongs_to :quorum_blastp_job end private def initialize(args) @algorithm = args[:search_tool] @id = args[:id] @log_directory = args[:log_directory] @tmp = args[:tmp_directory] @search_database = args[:search_database] @threads = args[:threads] @logger = Quorum::Logger.new(@log_directory) begin @job = QuorumJob.find(@id) rescue Exception => e @logger.log("ActiveRecord", e.message, 1) end @na_sequence = @job.na_sequence @aa_sequence = @job.aa_sequence ## Create for method invocation ## @job_association = "quorum_#{@algorithm}_job".to_sym @job_report_association = "quorum_#{@algorithm}_job_reports".to_sym @filter = @job.method(@job_association).call.filter @expectation = @job.method(@job_association).call.expectation @max_score = @job.method(@job_association).call.max_score @min_score = @job.method(@job_association).call.min_bit_score @gapped_alignments = @job.method(@job_association).call.gapped_alignments @gap_opening_penalty = @job.method(@job_association).call.gap_opening_penalty @gap_extension_penalty = @job.method(@job_association).call.gap_extension_penalty @db = @job.method(@job_association).call.blast_dbs.split(';') @db.map! { |d| File.join(@search_database, d) } @db = @db.join(' ') @hash = create_hash(@job.sequence) @tmp_files = File.join(@tmp, @hash) << "*" end # # Create a unique hash plus timestamp. # def create_hash(sequence) Digest::MD5.hexdigest(sequence).to_s + "-" + Time.now.to_f.to_s end # # Blastn command # def generate_blastn_cmd blastn = "blastn " << "-db \"#{@db}\" " << "-query #{@na_fasta} " << "-outfmt 5 " << "-num_threads #{@threads} " << "-evalue #{@expectation} " << "-max_target_seqs #{@max_score} " << "-out #{@out} " if @gapped_alignments blastn << "-gapopen #{@gap_opening_penalty} " blastn << "-gapextend #{@gap_extension_penalty} " else blastn << "-ungapped " end blastn << "-dust yes " if @filter blastn end # # Blastx command # def generate_blastx_cmd blastx = "blastx " << "-db \"#{@db}\" " << "-query #{@na_fasta} " << "-outfmt 5 " << "-num_threads #{@threads} " << "-evalue #{@expectation} " << "-max_target_seqs #{@max_score} " << "-out #{@out} " if @gapped_alignments blastx << "-gapopen #{@gap_opening_penalty} " blastx << "-gapextend #{@gap_extension_penalty} " else blastx << "-ungapped " end blastx << "-seg yes " if @filter blastx end # # Tblastn command # def generate_tblastn_cmd tblastn = "tblastn " << "-db \"#{@db}\" " << "-query #{@aa_fasta} " << "-outfmt 5 " << "-num_threads #{@threads} " << "-evalue #{@expectation} " << "-max_target_seqs #{@max_score} " << "-out #{@out} " if @gapped_alignments tblastn << "-gapopen #{@gap_opening_penalty} " tblastn << "-gapextend #{@gap_extension_penalty} " tblastn << "-comp_based_stats D " else tblastn << "-ungapped " tblastn << "-comp_based_stats F " end tblastn << "-seg yes " if @filter tblastn end # # Blastp command # def generate_blastp_cmd blastp = "blastp " << "-db \"#{@db}\" " << "-query #{@aa_fasta} " << "-outfmt 5 " << "-num_threads #{@threads} " << "-evalue #{@expectation} " << "-max_target_seqs #{@max_score} " << "-out #{@out} " if @gapped_alignments blastp << "-gapopen #{@gap_opening_penalty} " blastp << "-gapextend #{@gap_extension_penalty} " blastp << "-comp_based_stats D " else blastp << "-ungapped " blastp << "-comp_based_stats F " end blastp << "-seg yes " if @filter blastp end # # Generate Blast Command # def generate_blast_cmd @cmd = "" @na_fasta = File.join(@tmp, @hash + ".na.fa") @aa_fasta = File.join(@tmp, @hash + ".aa.fa") File.open(@na_fasta, "w") { |f| f << @na_sequence } File.open(@aa_fasta, "w") { |f| f << @aa_sequence } @out = File.join(@tmp, @hash + ".out.xml") File.new(@out, "w") case @algorithm when "blastn" @cmd << generate_blastn_cmd when "blastx" @cmd << generate_blastx_cmd when "tblastn" @cmd << generate_tblastn_cmd when "blastp" @cmd << generate_blastp_cmd end end # # Format Blast report hit_display_id and hit_def. # # For added flexibility, Quorum doesn't parse seqids or deflines. # Instead, format_hit splits the hit_def on whitespace and # reports hit_display_id as the first element and hit_def as the second. # def format_hit(str) hit_display_id = "" hit_def = "" hit = str.split(" ", 2) if hit.length < 2 hit_display_id = hit.first hit_def = "None" else hit_display_id = hit.first hit_def = hit.last end return hit_display_id, hit_def end # # Save Blast Job Report # # Hsps are only reported if a query hit against the Blast db. # Only save the @data if bit_score exists and it's > the user # defined minimum score. # # Set the attribute results to true for downstream processes. # def save_hsp_results if @data[:bit_score] && (@data[:bit_score].to_i > @min_score.to_i) @data[:results] = true @data["#{@algorithm}_job_id".to_sym] = @job.method(@job_association).call.job_id job_report = @job.method(@job_report_association).call.build(@data) unless job_report.save! @logger.log( "ActiveRecord", "Unable to save #{@algorithm} results to database.", 1, @tmp_files ) end end end # # Save empty Blast Job Report # # Set the attribute results to false for downstream processes. # def save_empty_results job_report = @job.method(@job_report_association).call.build( "#{@algorithm}_job_id" => @job.method(@job_association).call.job_id, "results" => false ) unless job_report.save! @logger.log( "ActiveRecord", "Unable to save #{@algorithm} results to database.", 1, @tmp_files ) end @logger.log( "NCBI Blast", "#{@algorithm} report empty.", 0, @tmp_files ) end # # Parse and save Blast results. # # Parse the Blast XML output and save results. # def parse_and_save_results @results = false # Did the xml contain results? if File.size(@out) > 0 report = Bio::Blast::XmlIterator.new(@out) report.to_enum.each do |iteration| @data = {} @data[:query] = iteration.query_def.split(" ").first @data[:query_len] = iteration.query_len iteration.each do |hit| @data[:hit_display_id], @data[:hit_def] = format_hit(hit.hit_def) @data[:hit_id] = hit.hit_id @data[:hit_accession] = hit.accession @data[:hit_len] = hit.len hit.each do |hsp| @data[:hsp_num] = hsp.hsp_num @data[:bit_score] = hsp.bit_score @data[:score] = hsp.score @data[:evalue] = hsp.evalue @data[:query_from] = hsp.query_from @data[:query_to] = hsp.query_to @data[:hit_from] = hsp.hit_from @data[:hit_to] = hsp.hit_to @data[:query_frame] = hsp.query_frame @data[:hit_frame] = hsp.hit_frame @data[:identity] = hsp.identity @data[:positive] = hsp.positive @data[:gaps] = hsp.gaps @data[:align_len] = hsp.align_len @data[:qseq] = hsp.qseq @data[:hseq] = hsp.hseq @data[:midline] = hsp.midline if @data[:bit_score] && (@data[:bit_score].to_i > @min_score.to_i) @results = true save_hsp_results end end end end end save_empty_results unless @results end # # Group record ids of hsps belonging to the same query and hit_accession. # def add_hps_groups_to_reports groups = {} query_hit_acc = @job.method( @job_report_association ).call.select("DISTINCT query, hit_accession") # Populate the hash of hashes with correct query and hit_accession. query_hit_acc.each do |q| groups[q.query] = {} end query_hit_acc.each do |q| groups[q.query][q.hit_accession] = [] end # Group record ids. reports = @job.method(@job_report_association).call.order("id ASC") reports.each do |r| if groups[r.query][r.hit_accession] groups[r.query][r.hit_accession] << r.id end end # Save hsp_group as a comma delimited string. reports.each do |r| if groups[r.query][r.hit_accession] if groups[r.query][r.hit_accession].length > 1 r.hsp_group = groups[r.query][r.hit_accession].join(",") unless r.save! @logger.log( "ActiveRecord", "Unable to save #{@algorithm} results to database.", 1, @tmp_files ) end end end end end # # Remove tmp files. # def remove_tmp_files `rm #{@tmp_files}` if @tmp_files end public # # Execute Blast on a given dataset. # def execute_blast generate_blast_cmd @logger.log("NCBI Blast", @cmd) system(@cmd) # Wrap these methods in a transaction to prevent premature return. @job.method(@job_report_association).call.transaction do parse_and_save_results add_hps_groups_to_reports end remove_tmp_files end end end end