def gather_guess_tax_at_each_rank(
gather_results,
num_hashes,
taxlist=lca_utils.taxlist(include_strain=False),
minimum_matches=3,
lowest_rank="genus"):
rank_results = []
prev_lineage = ""
top_lineage = ""
for rank in taxlist:
top_lineage, f_ident, f_major = gather_guess_tax_at_rank(
gather_results, num_hashes, rank, minimum_matches=minimum_matches)
# summarizing at a lower rank than exists will yield same result as prev. break!
if not top_lineage or top_lineage == prev_lineage:
break
rank_results.append(
RankSumGatherResult(lineage=top_lineage,
f_ident=f_ident,
f_major=f_major))
prev_lineage = top_lineage
if rank == lowest_rank:
break
return rank_results
def add_hashes_at_ranks(lineage_hashD, hashes_to_add, lineage, match_rank):
# first add full lineage
lineage_hashD[lineage].add_many(hashes_to_add)
for rank in lca_utils.taxlist(include_strain=False):
# TODO: add check to pop ONLY if needed (no need to pop at genus if lineage only has superk, phyl)
lin_at_rank = pop_to_rank(lineage, rank)
lineage_hashD[lin_at_rank].add_many(hashes_to_add)
if rank == match_rank:
break
return lineage_hashD
def test_searchfiles_contigs_just_gather(location):
prefix = os.path.join(location, "pref")
filelist = [
f"{prefix}.contigs.rankgather.csv", f"{prefix}.contigs.unmatched.fq"
]
sf = SearchFiles(prefix, search=True, gather=True)
# two minhashes, share ranks at phylum level
hashval = 12345678
ident1 = 'first'
mh1, sig1, lin1 = make_sig_and_lin([hashval], ident1, 'a;b;c')
hashval2 = 87654321
ident2 = 'second'
mh2, sig2, lin2 = make_sig_and_lin([hashval2], ident2, 'a;b;f')
# create lca_db w sigs
lca_db = LCA_Database(scaled=1, ksize=3)
lca_db.insert(sig1, ident=ident1)
lca_db.insert(sig2, ident=ident2)
# make lin_db
lin_db = LineageDB()
lin_db.insert(ident1, lin1)
lin_db.insert(ident2, lin2)
num_hashes = 2
# search with combined hashvals
search_mh = make_mh([hashval, hashval2])
gather_results = list(gather_at_rank(search_mh, lca_db, lin_db, "class"))
gather_rank_results=gather_guess_tax_at_each_rank(gather_results, num_hashes, minimum_matches=1, \
lowest_rank="class", \
taxlist=lca_utils.taxlist(include_strain=False))
#write search results
name = 'name'
seq_len = 6
for gres in gather_rank_results:
sf.write_result(gres, name, seq_len, result_type="rankgather")
sf.close()
# check results are in files
for f in filelist:
assert os.path.exists(f)
with open(f"{prefix}.contigs.rankgather.csv", "r") as gatherres:
this_gather_csvset = get_csv_set(gatherres)
with open(utils.get_testfile("test-data/test.contigs.rankgather.csv"),
"r") as searchres:
saved_gather_csvset = get_csv_set(searchres)
assert saved_gather_csvset == this_gather_csvset
def sort_by_rank_and_containment(summarized_results, match_rank):
sorted_results = []
# iterate superkingdom --> match_rank
for rank in lca_utils.taxlist(include_strain=False):
rank_res = summarized_results[rank]
rank_res.sort(key=itemgetter(1), reverse=True) # sort by containment
for (lin, containment, intersect_bp, match_sig) in rank_res:
sorted_results.append(
RankSumSearchResult(lineage=lin,
contained_at_rank=containment,
contained_bp=intersect_bp,
match=match_sig))
if rank == match_rank:
break
return sorted_results
def test_gather_guess_tax_at_each_rank_1():
#two minhashes, fully shared ranks
# first sig
hashval = 12345678
ident1 = 'first'
mh1, sig1, lin1 = make_sig_and_lin([hashval], ident1, 'a;b;c')
# second sig
hashval2 = 87654321
ident2 = 'second'
mh2, sig2, lin2 = make_sig_and_lin([hashval2], ident2, 'a;b;c')
# create lca_db w sigs
lca_db = LCA_Database(scaled=1, ksize=3)
lca_db.insert(sig1, ident=ident1)
lca_db.insert(sig2, ident=ident2)
# make lin_db
lin_db = LineageDB()
lin_db.insert(ident1, lin1)
lin_db.insert(ident2, lin2)
num_hashes = 2
superk_lin = lca_utils.make_lineage('a')
phylum_lin = lca_utils.make_lineage('a;b')
# search with combined hashvals
search_mh = make_mh([hashval, hashval2])
gather_results = list(gather_at_rank(search_mh, lca_db, lin_db, "class"))
rank_results=gather_guess_tax_at_each_rank(gather_results, num_hashes, minimum_matches=1, \
lowest_rank="class",
taxlist=lca_utils.taxlist(include_strain=False))
assert len(rank_results) == 3
assert rank_results[0] == RankSumGatherResult(lineage=superk_lin,
f_ident=1.0,
f_major=1.0)
assert rank_results[1] == RankSumGatherResult(lineage=phylum_lin,
f_ident=1.0,
f_major=1.0)
assert rank_results[2] == RankSumGatherResult(lineage=lin1,
f_ident=1.0,
f_major=1.0)
def test_gather_guess_tax_at_each_rank_3():
# two minhashes, totally distinct ranks
# first sig
hashval1 = 12345678
ident1 = 'first'
mh1, sig1, lin1 = make_sig_and_lin([hashval1], ident1, 'a;b;c')
# second sig
hashval2 = 87654321
ident2 = 'second'
mh2, sig2, lin2 = make_sig_and_lin([hashval2], ident2, 'd;e;f')
# create lca_db w sig1
lca_db = LCA_Database(scaled=1, ksize=3)
lca_db.insert(sig1, ident=ident1)
lca_db.insert(sig2, ident=ident2)
# next, make lin_db
lin_db = LineageDB()
lin_db.insert(ident1, lin1)
lin_db.insert(ident2, lin2)
num_hashes = 2
#winner seems to be def lineage.. will this remain true always?
superk_lin = lca_utils.make_lineage('d')
phylum_lin = lca_utils.make_lineage('d;e')
# search with combined hashvals
search_mh = make_mh([hashval1, hashval2])
gather_results = list(gather_at_rank(search_mh, lca_db, lin_db, "class"))
rank_results=gather_guess_tax_at_each_rank(gather_results, num_hashes, minimum_matches=1, \
lowest_rank="class",
taxlist=lca_utils.taxlist(include_strain=False))
assert len(rank_results) == 3
assert rank_results[0] == RankSumGatherResult(lineage=superk_lin,
f_ident=1.0,
f_major=0.5)
assert rank_results[1] == RankSumGatherResult(lineage=phylum_lin,
f_ident=1.0,
f_major=0.5)
assert rank_results[2] == RankSumGatherResult(lineage=lin2,
f_ident=1.0,
f_major=0.5)
def pop_to_rank(lin, rank):
"Remove lineage tuples from given lineage `lin` until `rank` is reached."
lin = list(lin)
txl = lca_utils.taxlist()
before_rank = []
for txl_rank in txl:
if txl_rank != rank:
before_rank.append(txl_rank)
else:
break
# are we already above rank?
if lin and lin[-1].rank in before_rank:
return tuple(lin)
while lin and lin[-1].rank != rank:
lin.pop()
return tuple(lin)
def make_lca_counts(dblist,
lowest_rank='phylum',
min_num=0,
min_hashes=5,
prefix='oddities'):
"""
Collect counts of all the LCAs in the list of databases.
"""
assert len(dblist) == 1
keep_ranks = ['root']
for rank in lca_utils.taxlist():
keep_ranks.append(rank)
if rank == lowest_rank:
break
print('keeping hashvals at following ranks:', keep_ranks)
print('min number of lineages:', min_num)
print('min number of shared hashes:', min_hashes)
print('---')
# gather all hashvalue assignments from across all the databases
assignments = defaultdict(set)
for lca_db in dblist:
for hashval, idx_list in lca_db.hashval_to_idx.items():
if min_num and len(idx_list) < min_num:
continue
for idx in idx_list:
lid = lca_db.idx_to_lid.get(idx)
if lid is not None:
lineage = lca_db.lid_to_lineage[lid]
assignments[hashval].add(lineage)
# now convert to trees -> do LCA & counts
counts = defaultdict(int)
mixdict = defaultdict(set)
for hashval, lineages in assignments.items():
# for each list of tuple_info [(rank, name), ...] build
# a tree that lets us discover lowest-common-ancestor.
debug("{}", lineages)
tree = lca_utils.build_tree(lineages)
# now find either a leaf or the first node with multiple
# children; that's our lowest-common-ancestor node.
lca, reason = lca_utils.find_lca(tree)
# find cross-superkingdom hashes, and record combinations of lineages
# that have them.
rank = 'root'
if lca:
rank = lca[-1].rank
if rank in keep_ranks:
xx = []
for lineage in lineages:
xx.append(tuple(lineage))
xx = tuple(xx)
mixdict[xx].add(hashval)
counts[lca] += 1
# sort on number of confused hash vals by combination of lineages.
mixdict_items = list(mixdict.items())
mixdict_items.sort(key=lambda x: -len(x[1]))
confused_hashvals = set()
fp = open(prefix + '.csv', 'wt')
w = csv.writer(fp)
w.writerow([
'cluster', 'num_lineages', 'shared_kmers', 'ksize', 'rank', 'lca',
'ident1', 'lineage1', 'ident2', 'lineage2'
])
#
# find candidate lineages, then evaluate pairwise intersections.
#
for cluster_n, (lineages, hashvals) in enumerate(mixdict_items):
# insist on more than N hash vals
if len(hashvals) < min_hashes:
continue
# display summary:
print('cluster {} has {} assignments for {} hashvals / {} bp'.format(
cluster_n, len(lineages), len(hashvals),
dblist[0].scaled * len(hashvals)))
confused_hashvals.update(hashvals)
tree = lca_utils.build_tree(lineages)
lca, reason = lca_utils.find_lca(tree)
if lca:
rank = lca[-1].rank
else:
rank = 'root'
print(' rank & lca:', rank, lca_utils.display_lineage(lca))
# for lineage_n, lineage in enumerate(lineages):
# print('* ', lca_utils.display_lineage(lineage))
# now, identify all members of these lineages by their index:
all_idxs = []
for lineage_n, lineage in enumerate(lineages):
lids = dblist[0].lineage_to_lids[lineage]
for lid in lids:
idxs = dblist[0].lid_to_idx[lid]
all_idxs.extend(idxs)
for idx in idxs:
ident = dblist[0].idx_to_ident[idx]
# run through and look at all pairs of genomes in these lineages;
# filter so that we're comparing across lineages with the right
# LCA, and with significant intersection.
pair_n = 0
candidates = []
for i in range(len(all_idxs)):
idx1 = all_idxs[i]
lid1 = dblist[0].idx_to_lid[idx1]
lin1 = dblist[0].lid_to_lineage[lid1]
for j in range(i):
idx2 = all_idxs[j]
lid2 = dblist[0].idx_to_lid[idx2]
lin2 = dblist[0].lid_to_lineage[lid2]
ident1 = dblist[0].idx_to_ident[idx1]
ident2 = dblist[0].idx_to_ident[idx2]
debug("{} x {}", ident1, ident2)
this_tree = lca_utils.build_tree([lin1, lin2])
this_lca, this_reason = lca_utils.find_lca(this_tree)
# weed out pairs that don't have the desired intersection
if lca != this_lca:
continue
mh1 = dblist[0]._signatures[idx1]
mh2 = dblist[0]._signatures[idx2]
mins1 = set(mh1.get_mins())
mins2 = set(mh2.get_mins())
intersect_size = len(mins1.intersection(mins2))
# weed out pairs that don't have enough k-mer intersection
if intersect_size < min_hashes:
continue
candidates.append(
(pair_n, ident1, lin1, ident2, lin2, intersect_size))
# write summary to CSV for find-oddities-examine.py to use
w.writerow([
'cluster{}.{}'.format(cluster_n, pair_n),
len(lineages), intersect_size * dblist[0].scaled,
dblist[0].ksize, rank,
lca_utils.display_lineage(lca), ident1,
lca_utils.display_lineage(lin1), ident2,
lca_utils.display_lineage(lin2)
])
pair_n += 1
print(' Candidate genome pairs for these lineages:')
for pair_n, ident1, lin1, ident2, lin2, intersection_size in candidates:
print(' cluster.pair {}.{} share {} bases'.format(
cluster_n, pair_n, intersection_size * dblist[0].scaled))
print(' - {} ({})'.format(ident1,
lca_utils.display_lineage(lin1)))
print(' - {} ({})'.format(ident2,
lca_utils.display_lineage(lin2)))
print('')
print('')
return counts, confused_hashvals
def test_taxlist_2():
assert list(taxlist(include_strain=False)) == ['superkingdom', 'phylum', 'class', 'order', 'family', 'genus', 'species']
def test_taxlist_1():
assert list(taxlist()) == ['superkingdom', 'phylum', 'class', 'order', 'family', 'genus', 'species', 'strain']
def main(args):
"Main entry point for scripting. Use cmdline for command line entry."
genomebase = os.path.basename(args.genome)
match_rank = 'genus'
# load taxonomy CSV
tax_assign, _ = load_taxonomy_assignments(args.lineages_csv,
start_column=2)
print(f'loaded {len(tax_assign)} tax assignments.')
# load the genome signature
genome_sig = sourmash.load_one_signature(args.genome_sig,
select_moltype=args.alphabet,
ksize=args.ksize)
# load all of the matches from search --containment in the database
with open(args.matches_sig, 'rt') as fp:
try:
siglist = list(
sourmash.load_signatures(fp, do_raise=True, quiet=False))
except sourmash.exceptions.SourmashError:
siglist = []
print(f"loaded {len(siglist)} matches from '{args.matches_sig}'")
# Hack for examining members of our search database: remove exact matches.
new_siglist = []
for ss in siglist:
if genome_sig.similarity(ss) == 1.0:
print(f'removing an identical match: {ss.name()}')
else:
new_siglist.append(ss)
siglist = new_siglist
if not siglist:
# write empty files so snakemake workflows don't complain; exit.
print('no non-identical matches for this genome, exiting.')
if not args.no_search_contigs:
sf = SearchFiles(args.output_prefix,
not args.no_search,
args.gather,
contigs=True)
sf.close()
if args.search_genome:
gf = SearchFiles(args.output_prefix,
not args.no_search,
args.gather,
contigs=False)
gf.close()
return 0
# construct a template minhash object that we can use to create new 'uns
empty_mh = siglist[0].minhash.copy_and_clear()
ksize = empty_mh.ksize
scaled = empty_mh.scaled
moltype = empty_mh.moltype
# create empty LCA database to populate...
lca_db = LCA_Database(ksize=ksize, scaled=scaled, moltype=moltype)
lin_db = LineageDB()
# ...with specific matches.
for ss in siglist:
ident = get_ident(ss)
lineage = tax_assign[ident]
lca_db.insert(ss, ident=ident)
lin_db.insert(ident, lineage)
print(f'loaded {len(siglist)} signatures & created LCA Database')
print('')
print(f'reading contigs from {genomebase}')
screed_iter = screed.open(args.genome)
genome_len = 0
if not args.no_search_contigs:
sf = SearchFiles(args.output_prefix,
not args.no_search,
args.gather,
contigs=True)
for n, record in enumerate(screed_iter):
# look at each contig individually
mh = empty_mh.copy_and_clear()
mh.add_sequence(record.sequence, force=True)
# search, optionally aggregate matched hashes to get containment at rank
seq_len = len(record.sequence)
genome_len += seq_len
num_hashes = len(mh.hashes)
if not args.no_search:
search_results, search_rank_results = search_containment_at_rank(
mh, lca_db, lin_db, match_rank)
if not search_results:
# write to unclassified
sf.unmatched.write(">" + record.name + "\n" +
record.sequence + "\n")
continue # if no search results, don't bother with gather
else:
# first, print normal search --containment results
for sr in search_results:
sf.write_result(sr,
record.name,
seq_len,
result_type="search")
# now, print containment at rank results
for sr in search_rank_results:
sf.write_result(sr,
record.name,
seq_len,
result_type="ranksearch")
if args.gather:
# first, gather at match rank (default genus)
gather_results = list(
gather_at_rank(mh, lca_db, lin_db, match_rank))
# write standard gather_results?
if not gather_results:
# write to unclassified. should only get here if no search OR gather results
sf.unmatched.write(">" + record.name + "\n" +
record.sequence + "\n")
else:
# next, summarize at higher ranks
gather_taxonomy_per_rank = gather_guess_tax_at_each_rank(gather_results, num_hashes, \
minimum_matches=args.gather_min_matches, \
lowest_rank=match_rank, \
taxlist=lca_utils.taxlist(include_strain=False))
#results = list of RankSumGatherResult = namedtuple('RankSumGatherResult', 'lineage, f_ident, f_major')
# write taxonomy out
for gr in gather_taxonomy_per_rank:
sf.write_result(gr,
record.name,
seq_len,
result_type="rankgather")
print(f"Processed {n+1} contigs.")
# close contig files
sf.close()
if args.search_genome:
gf = SearchFiles(args.output_prefix,
not args.no_search,
args.gather,
contigs=False)
# MAG workflow
entire_mh = genome_sig.minhash
genome_name = genome_sig.name()
num_hashes = len(entire_mh.hashes)
if not genome_len:
for record in screed_iter:
genome_len += len(record.sequence)
if not args.no_search:
#results are guaranteed, otherwise would have exited before searching
search_results, search_rank_results = search_containment_at_rank(
entire_mh, lca_db, lin_db, match_rank)
for sr in search_results:
gf.write_result(sr,
genome_name,
genome_len,
result_type="search")
for sr in search_rank_results:
gf.write_result(sr,
genome_name,
genome_len,
result_type="ranksearch")
if args.gather:
gather_results = list(
gather_at_rank(entire_mh, lca_db, lin_db, match_rank))
# next, summarize at higher ranks
gather_taxonomy_per_rank = gather_guess_tax_at_each_rank(gather_results, num_hashes, \
minimum_matches=args.gather_min_matches, \
lowest_rank=match_rank, \
taxlist=lca_utils.taxlist(include_strain=False))
for gather_res in gather_taxonomy_per_rank:
gf.write_result(gather_res,
genome_name,
genome_len,
result_type="rankgather")
# close genome files
gf.close()
return 0