# encoding: utf-8 require 'bio' require 'forwardable' module Cheripic # Custom error handling for ContigPileup class class ContigPileupsError < CheripicError; end # A ContigPileup object for each contig from assembly that stores # pileup file information and variants are selected from analysis of pileup files # selected variants from pileup files is stored as hashes # # @!attribute [rw] id # @return [String] id of the contig in assembly taken from fasta file # @!attribute [rw] mut_bulk # @return [Hash] a hash of variant positions from mut_bulk as keys and pileup info as values # @!attribute [rw] bg_bulk # @return [Hash] a hash of variant positions from bg_bulk as keys and pileup info as values # @!attribute [rw] mut_parent # @return [Hash] a hash of variant positions from mut_parent as keys and pileup info as values # @!attribute [rw] bg_parent # @return [Hash] a hash of variant positions from bg_parent as keys and pileup info as values # @!attribute [rw] parent_hemi # @return [Hash] a hash of hemi-variant positions as keys and bfr calculated from parent bulks as values class ContigPileups include Enumerable extend Forwardable def_delegators :@mut_bulk, :each, :each_key, :each_value, :length, :[], :store def_delegators :@bg_bulk, :each, :each_key, :each_value, :length, :[], :store def_delegators :@mut_parent, :each, :each_key, :each_value, :length, :[], :store def_delegators :@bg_parent, :each, :each_key, :each_value, :length, :[], :store attr_accessor :id, :parent_hemi attr_accessor :mut_bulk, :bg_bulk, :mut_parent, :bg_parent # creates a ContigPileup object using fasta entry id # @param fasta [String] a contig id from fasta entry def initialize (fasta) @id = fasta @mut_bulk = {} @bg_bulk = {} @mut_parent = {} @bg_parent = {} @parent_hemi = {} end # bulk pileups are compared and variant positions are selected # @return [Array] variant positions are stored in hashes # for homozygous, heterozygous and hemi-variant positions def bulks_compared @hm_pos = {} @ht_pos = {} @hemi_pos = {} @mut_bulk.each_key do | pos | if Options.polyploidy and @parent_hemi.key?(pos) bg_bases = '' if @bg_bulk.key?(pos) bg_bases = @bg_bulk[pos].var_base_frac end mut_bases = @mut_bulk[pos].var_base_frac bfr = Bfr.get_bfr(mut_bases, bg_bases) @hemi_pos[pos] = bfr else self.compare_pileup(pos) end end [@hm_pos, @ht_pos, @hemi_pos] end # mut_bulk and bg_bulk pileups are compared at selected position of the contig. # Empty hash results from position below selected coverage # or bases freq below noise and such positions are deleted. # @param pos [Integer] position in the contig # stores variant type, position and allele fraction to either @hm_pos or @ht_pos hashes def compare_pileup(pos) base_hash = @mut_bulk[pos].var_base_frac base_hash.delete(:ref) return nil if base_hash.empty? # we could ignore complex loci or # take the variant type based on predominant base if base_hash.length > 1 fraction = base_hash.values.max mut_type = var_mode(fraction) else fraction = base_hash[base_hash.keys[0]] mut_type = var_mode(fraction) end if @bg_bulk.key?(pos) bg_type = bg_bulk_var(pos) mut_type = compare_var_type(mut_type, bg_type) end unless mut_type == nil categorise_pos(mut_type, pos, fraction) end end # Categorizes variant zygosity based on the allele fraction provided. # Uses lower and upper limit set for heterozygosity in the options. # @note consider increasing the range of heterozygosity limits for RNA-seq data # @param fraction [Float] allele fraction # @return [Symbol] of either :het or :hom to represent heterozygous or homozygous respectively def var_mode(fraction) ht_low = Options.htlow ht_high = Options.hthigh mode = '' if fraction.between?(ht_low, ht_high) mode = :het elsif fraction > ht_high mode = :hom end mode end # Simple comparison of variant type of mut and bg bulks at a position # If both bulks have homozygous variant at selected position then it is ignored # @param muttype [Symbol] values are either :hom or :het # @param bgtype [Symbol] values are either :hom or :het # @return [Symbol] variant mode of the mut bulk (:hom or :het) at the position or nil def compare_var_type(muttype, bgtype) if muttype == :hom and bgtype == :hom nil else muttype end end # Method to extract var_mode from pileup information at a position in contig # # @param pos [Integer] position in the contig # @return [Symbol] variant mode of the background bulk (:hom or :het) at the position def bg_bulk_var(pos) bg_base_hash = @bg_bulk[pos].var_base_frac bg_base_hash.delete(:ref) return nil if bg_base_hash.empty? if bg_base_hash.length > 1 # taking only var mode var_mode(bg_base_hash.values.max) else # taking only var mode var_mode(bg_base_hash[bg_base_hash.keys[0]]) end end # method stores pos as key and allele fraction as value # to @hm_pos or @ht_pos hash based on variant type # @param var_type [Symbol] values are either :hom or :het # @param pos [Integer] position in the contig # @param ratio [Float] allele fraction def categorise_pos(var_type, pos, ratio) if var_type == :hom @hm_pos[pos] = ratio elsif var_type == :het @ht_pos[pos] = ratio end end # Compares parental pileups for the contig and identify position # that indicate variants from homelogues called hemi-snps # and calculates bulk frequency ratio (bfr) # @return [Hash] parent_hemi hash with position as key and bfr as value def hemisnps_in_parent # mark all the hemi snp based on both parents self.mut_parent.each_key do |pos| mut_parent_frac = @mut_parent[pos].var_base_frac if self.bg_parent.key?(pos) bg_parent_frac = @bg_parent[pos].var_base_frac bfr = Bfr.get_bfr(mut_parent_frac, bg_parent_frac) @parent_hemi[pos] = bfr self.bg_parent.delete(pos) else bfr = Bfr.get_bfr(mut_parent_frac) @parent_hemi[pos] = bfr end end # now include all hemi snp unique to background parent self.bg_parent.each_key do |pos| unless @parent_hemi.key?(pos) bg_parent_frac = @bg_parent[pos].var_base_frac bfr = Bfr.get_bfr(bg_parent_frac) @parent_hemi[pos] = bfr end end end end end