def test_nucleotide_search_unaligned_reads_read_count_aligned_subject_coverage(self):
"""
Test the unaligned reads and the store alignments
Test with a bowtie2/sam output file
Test for aligned read counts
Test with subject coverage filtering
"""
# create a set of alignments
alignments=store.Alignments()
unaligned_reads_store=store.Reads()
# turn off subject filtering
config.nucleotide_query_coverage_threshold = 0
# read in the aligned and unaligned reads
[unaligned_reads_file_fasta, reduced_aligned_reads_file] = nucleotide.unaligned_reads(
cfg.sam_file_unaligned_reads, alignments, unaligned_reads_store, keep_sam=True)
# reset subject filtering
config.nucleotide_query_coverage_threshold = self.default_nucleotide_query_coverage_threshold
# remove temp files
utils.remove_temp_file(unaligned_reads_file_fasta)
utils.remove_temp_file(reduced_aligned_reads_file)
# check the aligned reads count
self.assertEqual(len(alignments.get_hit_list()),cfg.sam_file_unaligned_reads_total_aligned_subject_coverage)
def test_nucleotide_search_unaligned_reads_output_fasta_format(self):
"""
Test the unaligned reads and the store alignments
Test with a bowtie2/sam output file
Test output file is of fasta format
Test sam file is not removed
"""
# create a set of alignments
alignments=store.Alignments()
unaligned_reads_store=store.Reads()
# turn off query/subject filtering
config.nucleotide_subject_coverage_threshold = 0
config.nucleotide_query_coverage_threshold = 0
# read in the aligned and unaligned reads
[unaligned_reads_file_fasta, reduced_aligned_reads_file] = nucleotide.unaligned_reads(
cfg.sam_file_unaligned_reads, alignments, unaligned_reads_store, keep_sam=True)
# reset query/subject filtering
config.nucleotide_subject_coverage_threshold = self.default_nucleotide_subject_coverage_threshold
config.nucleotide_query_coverage_threshold = self.default_nucleotide_query_coverage_threshold
# check for fasta output file format
file_format=utilities.determine_file_format(unaligned_reads_file_fasta)
self.assertEqual("fasta",file_format)
# remove temp files
utils.remove_temp_file(unaligned_reads_file_fasta)
utils.remove_temp_file(reduced_aligned_reads_file)
def test_nucleotide_search_unaligned_reads_output_blast_format(self):
"""
Test the unaligned reads and the store alignments
Test with a bowtie2/sam output file
Test the aligned reads file created is of the blastm8 format
"""
# create a set of alignments
alignments=store.Alignments()
unaligned_reads_store=store.Reads()
# turn off query/subject filtering
config.nucleotide_subject_coverage_threshold = 0
config.nucleotide_query_coverage_threshold = 0
config.file_basename="TEST"
# read in the aligned and unaligned reads
[unaligned_reads_file_fasta, reduced_aligned_reads_file] = nucleotide.unaligned_reads(
cfg.sam_file_annotations, alignments, unaligned_reads_store, keep_sam=True)
# reset query/subject filtering
config.nucleotide_subject_coverage_threshold = self.default_nucleotide_subject_coverage_threshold
config.nucleotide_query_coverage_threshold = self.default_nucleotide_query_coverage_threshold
# test file is of the blastm8 format
file_format=utilities.determine_file_format(reduced_aligned_reads_file)
# remove temp files
utils.remove_temp_file(unaligned_reads_file_fasta)
utils.remove_temp_file(reduced_aligned_reads_file)
self.assertEqual(file_format,"blastm8")
def test_nucleotide_search_unaligned_reads_annotations_reference(self):
"""
Test the unaligned reads and the store alignments
Test with a bowtie2/sam output file
Test the different annotation formats are recognized for reference
"""
# create a set of alignments
alignments=store.Alignments()
unaligned_reads_store=store.Reads()
# turn off query/subject filtering
config.nucleotide_subject_coverage_threshold = 0
config.nucleotide_query_coverage_threshold = 0
# read in the aligned and unaligned reads
[unaligned_reads_file_fasta, reduced_aligned_reads_file] = nucleotide.unaligned_reads(
cfg.sam_file_annotations, alignments, unaligned_reads_store, keep_sam=True)
# reset query/subject filtering
config.nucleotide_subject_coverage_threshold = self.default_nucleotide_subject_coverage_threshold
config.nucleotide_query_coverage_threshold = self.default_nucleotide_query_coverage_threshold
# remove temp files
utils.remove_temp_file(unaligned_reads_file_fasta)
utils.remove_temp_file(reduced_aligned_reads_file)
# two of the hits should be for gene "UniRef50"
hits=alignments.hits_for_gene("UniRef50")
self.assertEqual(len(hits),2)
def test_nucleotide_search_unaligned_reads_annotations_bug(self):
"""
Test the unaligned reads and the store alignments
Test with a bowtie2/sam output file
Test the different annotation formats are recognized for bug
"""
# create a set of alignments
alignments=store.Alignments()
unaligned_reads_store=store.Reads()
# turn off query/subject filtering
config.nucleotide_subject_coverage_threshold = 0
config.nucleotide_query_coverage_threshold = 0
# read in the aligned and unaligned reads
[unaligned_reads_file_fasta, reduced_aligned_reads_file] = nucleotide.unaligned_reads(
cfg.sam_file_annotations, alignments, unaligned_reads_store, keep_sam=True)
# reset query/subject filtering
config.nucleotide_subject_coverage_threshold = self.default_nucleotide_subject_coverage_threshold
config.nucleotide_query_coverage_threshold = self.default_nucleotide_query_coverage_threshold
# remove temp files
utils.remove_temp_file(unaligned_reads_file_fasta)
utils.remove_temp_file(reduced_aligned_reads_file)
# there should be one bug which is unclassified
self.assertEqual(alignments.bug_list(),["unclassified"])
def test_translated_search_unaligned_reads_annotations_bug(self):
"""
Test the unaligned reads and the store alignments
Test with a rapsearch2 output file
Test the different annotation formats are recognized for bug
Test without the coverage filter
"""
# create a set of alignments
alignments = store.Alignments()
unaligned_reads_store = store.Reads()
# set the coverage threshold to zero so as to not test with filter on
current_coverage_threshold = config.translated_subject_coverage_threshold
config.translated_subject_coverage_threshold = 0
# load the rapsearch2 output with the unaligned reads function
unaligned_file_fasta = translated.unaligned_reads(
unaligned_reads_store, cfg.rapsearch_file_annotations, alignments)
# remove temp file
utils.remove_temp_file(unaligned_file_fasta)
# reset the coverage threshold
config.translated_subject_coverage_threshold = current_coverage_threshold
# there should be one bug name and the other should be unclassified
self.assertEqual(
sorted(alignments.bug_list()),
sorted([
"g__Bacteroides.s__Bacteroides_xylanisolvens", "unclassified"
]))
def test_nucleotide_search_unaligned_reads_read_count_unaligned_minimize_memory_use(self):
"""
Test the unaligned reads and the store alignments
Test with a bowtie2/sam output file
Test for unaligned read counts
Test with minimize memory use
"""
# create a set of alignments
alignments=store.Alignments()
unaligned_reads_store=store.Reads(minimize_memory_use=True)
# turn off query/subject filtering
config.nucleotide_subject_coverage_threshold = 0
config.nucleotide_query_coverage_threshold = 0
# read in the aligned and unaligned reads
[unaligned_reads_file_fasta, reduced_aligned_reads_file] = nucleotide.unaligned_reads(
cfg.sam_file_unaligned_reads, alignments, unaligned_reads_store, keep_sam=True)
# reset query/subject filtering
config.nucleotide_subject_coverage_threshold = self.default_nucleotide_subject_coverage_threshold
config.nucleotide_query_coverage_threshold = self.default_nucleotide_query_coverage_threshold
# remove temp files
utils.remove_temp_file(unaligned_reads_file_fasta)
utils.remove_temp_file(reduced_aligned_reads_file)
# check the unaligned reads count
self.assertEqual(unaligned_reads_store.count_reads(),cfg.sam_file_unaligned_reads_total_unaligned)
def test_translated_search_unaligned_reads_annotations_reference(self):
"""
Test the unaligned reads and the store alignments
Test with a rapsearch2 output file
Test the different annotation formats are recognized for reference
Test without the coverage filter
"""
# create a set of alignments
alignments = store.Alignments()
unaligned_reads_store = store.Reads()
# set the coverage threshold to zero so as to not test with filter on
current_coverage_threshold = config.translated_subject_coverage_threshold
config.translated_subject_coverage_threshold = 0
# load the rapsearch2 output with the unaligned reads function
unaligned_file_fasta = translated.unaligned_reads(
unaligned_reads_store, cfg.rapsearch_file_annotations, alignments)
# remove temp file
utils.remove_temp_file(unaligned_file_fasta)
# reset the coverage threshold
config.translated_subject_coverage_threshold = current_coverage_threshold
# three of the hits should be for gene "UniRef50"
hits = alignments.hits_for_gene("UniRef50")
self.assertEqual(len(hits), 3)
def test_translated_search_unaligned_reads_blastm8(self):
"""
Test the unaligned reads and the store alignments
Test with a blastm8-like output file
Test with empty reads structure
Test that function does not require gene lengths in reference id
Test without the coverage filter
"""
# create a set of alignments
alignments = store.Alignments()
# set the coverage threshold to zero so as to not test with filter on
current_coverage_threshold = config.translated_subject_coverage_threshold
config.translated_subject_coverage_threshold = 0
# load the blastm8-like output
file_handle = open(cfg.rapsearch2_output_file_without_header)
for line in file_handle:
if not re.search("^#", line):
data = line.strip().split(config.blast_delimiter)
referenceid = data[config.blast_reference_index]
queryid = data[config.blast_query_index]
identity = float(data[config.blast_identity_index])
alignment_length = float(
data[config.blast_aligned_length_index])
alignments.add(referenceid, 0, queryid,
identity / 100.0 * alignment_length,
"unclassified", alignment_length)
file_handle.close()
alignments_test = store.Alignments()
unaligned_reads_store = store.Reads()
# load the blastm8-like output with the unaligned reads function
unaligned_file_fasta = translated.unaligned_reads(
unaligned_reads_store, cfg.rapsearch2_output_file_without_header,
alignments_test)
# remove temp file
utils.remove_temp_file(unaligned_file_fasta)
# reset the coverage threshold
config.translated_subject_coverage_threshold = current_coverage_threshold
# check the values are unchanged
self.assertEqual(sorted(alignments.get_hit_list()),
sorted(alignments_test.get_hit_list()))
def test_nucleotide_search_unaligned_reads_annotations_gene_length(self):
"""
Test the unaligned reads and the store alignments
Test with a bowtie2/sam output file
Test the different annotation formats are recognized for gene length
Test the gene length uses the read length from the sam file
"""
# create a set of alignments
alignments=store.Alignments()
unaligned_reads_store=store.Reads()
# turn off query/subject filtering
config.nucleotide_subject_coverage_threshold = 0
config.nucleotide_query_coverage_threshold = 0
# read in the aligned and unaligned reads
[unaligned_reads_file_fasta, reduced_aligned_reads_file] = nucleotide.unaligned_reads(
cfg.sam_file_annotations, alignments, unaligned_reads_store, keep_sam=True)
# reset query/subject filtering
config.nucleotide_subject_coverage_threshold = self.default_nucleotide_subject_coverage_threshold
config.nucleotide_query_coverage_threshold = self.default_nucleotide_query_coverage_threshold
# remove temp files
utils.remove_temp_file(unaligned_reads_file_fasta)
utils.remove_temp_file(reduced_aligned_reads_file)
# there should be 4 hits identified
all_hits=alignments.get_hit_list()
self.assertEqual(len(all_hits),4)
# check for set and default gene lengths
read_length = 151
expected_length_uniref50 = (abs(2000 - read_length)+1)/1000.0
expected_length_other = (abs(1000 - read_length)+1)/1000.0
for hit in all_hits:
query, bug, reference, score, length = hit
if reference == "UniRef50":
self.assertEqual(length,expected_length_uniref50)
else:
self.assertEqual(length,expected_length_other)
def test_translated_search_unaligned_reads_annotations_gene_length(self):
"""
Test the unaligned reads and the store alignments
Test with a rapsearch2 output file
Test the different annotation formats are recognized for gene length
Test without the coverage filter
"""
# create a set of alignments
alignments = store.Alignments()
unaligned_reads_store = store.Reads()
# set the coverage threshold to zero so as to not test with filter on
current_coverage_threshold = config.translated_subject_coverage_threshold
config.translated_subject_coverage_threshold = 0
# load the rapsearch2 output with the unaligned reads function
unaligned_file_fasta = translated.unaligned_reads(
unaligned_reads_store, cfg.rapsearch_file_annotations, alignments)
# remove temp file
utils.remove_temp_file(unaligned_file_fasta)
# reset the coverage threshold
config.translated_subject_coverage_threshold = current_coverage_threshold
# there should be 4 hits identified
all_hits = alignments.get_hit_list()
self.assertEqual(len(all_hits), 4)
# check for set and default gene lengths
read_length = 50
expected_length_uniref50 = (abs(2000 - read_length) + 1) / 1000.0
expected_length_other = (abs(1000 - read_length) + 1) / 1000.0
# check for set and default gene lengths
for hit in all_hits:
query, bug, reference, score, length = hit
if reference == "UniRef50":
self.assertEqual(length, expected_length_uniref50)
else:
self.assertEqual(length, expected_length_other)
def test_nucleotide_search_unaligned_reads_read_count_aligned_evalue_threshold(self):
"""
Test the unaligned reads and the store alignments
Test with a bowtie2/sam output file
Test for aligned read counts
Test the evalue threshold does not filter alignments
"""
# create a set of alignments
alignments=store.Alignments()
unaligned_reads_store=store.Reads()
# turn off query/subject filtering
config.nucleotide_subject_coverage_threshold = 0
config.nucleotide_query_coverage_threshold = 0
# update the evalue threshold to a number less than those for the alignment file
original_evalue_threshold=config.evalue_threshold
config.evalue_threshold=1e-15
# read in the aligned and unaligned reads
[unaligned_reads_file_fasta, reduced_aligned_reads_file] = nucleotide.unaligned_reads(
cfg.sam_file_unaligned_reads, alignments, unaligned_reads_store, keep_sam=True)
# reset query/subject filtering
config.nucleotide_subject_coverage_threshold = self.default_nucleotide_subject_coverage_threshold
config.nucleotide_query_coverage_threshold = self.default_nucleotide_query_coverage_threshold
# remove temp files
utils.remove_temp_file(unaligned_reads_file_fasta)
utils.remove_temp_file(reduced_aligned_reads_file)
# reset the evalue threshold back to the original
config.evalue_threshold=original_evalue_threshold
# check the aligned reads count (all reads should be aligned even though they do not
# meet the threshold as the evalue threshold is not applied for this type of alignment)
self.assertEqual(len(alignments.get_hit_list()),cfg.sam_file_unaligned_reads_total_aligned)
def test_nucleotide_search_unaligned_reads_read_count_aligned_identity_threshold(self):
"""
Test the unaligned reads and the store alignments
Test with a bowtie2/sam output file
Test for aligned read counts
Test the identity threshold does filter alignments
"""
# create a set of alignments
alignments=store.Alignments()
unaligned_reads_store=store.Reads()
# turn off query/subject filtering
config.nucleotide_subject_coverage_threshold = 0
config.nucleotide_query_coverage_threshold = 0
# update the identity threshold to a number larger than those in the alignments
original_identity_threshold=config.identity_threshold
config.identity_threshold=101.0
# read in the aligned and unaligned reads
[unaligned_reads_file_fasta, reduced_aligned_reads_file] = nucleotide.unaligned_reads(
cfg.sam_file_unaligned_reads, alignments, unaligned_reads_store, keep_sam=True)
# reset query/subject filtering
config.nucleotide_subject_coverage_threshold = self.default_nucleotide_subject_coverage_threshold
config.nucleotide_query_coverage_threshold = self.default_nucleotide_query_coverage_threshold
# remove temp files
utils.remove_temp_file(unaligned_reads_file_fasta)
utils.remove_temp_file(reduced_aligned_reads_file)
# reset the identity threshold back to the original
config.identity_threshold=original_identity_threshold
# check the aligned reads count (it should be two as both should pass the threshold)
self.assertEqual(len(alignments.get_hit_list()),2)
def test_nucleotide_search_unaligned_reads_scores(self):
"""
Test the unaligned reads and the store alignments
Test with a bowtie2/sam output file
Test the scores are based on percent identities
"""
# create a set of alignments
alignments=store.Alignments()
unaligned_reads_store=store.Reads()
# turn off query/subject filtering
config.nucleotide_subject_coverage_threshold = 0
config.nucleotide_query_coverage_threshold = 0
# read in the aligned and unaligned reads
[unaligned_reads_file_fasta, reduced_aligned_reads_file] = nucleotide.unaligned_reads(
cfg.sam_file_annotations, alignments, unaligned_reads_store, keep_sam=True)
# reset query/subject filtering
config.nucleotide_subject_coverage_threshold = self.default_nucleotide_subject_coverage_threshold
config.nucleotide_query_coverage_threshold = self.default_nucleotide_query_coverage_threshold
# remove temp files
utils.remove_temp_file(unaligned_reads_file_fasta)
utils.remove_temp_file(reduced_aligned_reads_file)
# there should be 4 hits identified
all_hits=alignments.get_hit_list()
# check for set and default gene lengths
expected_score=math.pow(151.0, config.match_power)
for hit in all_hits:
query, bug, reference, score, length = hit
self.assertEqual(score,expected_score)
def test_translated_search_unaligned_reads_blastm8_coverage_filter(self):
"""
Test the unaligned reads and the store alignments
Test with a blastm8-like output file
Test with empty reads structure
Test that function does not require gene lengths in reference id
Test with the coverage filter
Test with query length annotations
Test that an alignment with query start larger than query end is not filtered
"""
# create a set of alignments
alignments = store.Alignments()
# set the coverage threshold to a small value so as to have some alignments pass
current_coverage_threshold = config.translated_subject_coverage_threshold
config.translated_subject_coverage_threshold = 0.50
# get the set of allowed proteins
allowed_proteins = blastx_coverage.blastx_coverage(
cfg.rapsearch2_output_file_without_header_coverage,
config.translated_subject_coverage_threshold, alignments, True)
# load the blastm8-like output
file_handle = open(cfg.rapsearch2_output_file_without_header_coverage)
for line in file_handle:
if not re.search("^#", line):
data = line.strip().split(config.blast_delimiter)
referenceid = data[config.blast_reference_index]
gene, length, bug = alignments.process_reference_annotation(
referenceid)
queryid, query_length = utilities.get_length_annotation(
data[config.blast_query_index])
identity = float(data[config.blast_identity_index])
alignment_length = float(
data[config.blast_aligned_length_index])
if gene in allowed_proteins:
alignments.add(gene, length, queryid,
identity / 100.0 * alignment_length, bug,
alignment_length)
file_handle.close()
alignments_test = store.Alignments()
unaligned_reads_store = store.Reads()
# load the blastm8-like output with the unaligned reads function
unaligned_file_fasta = translated.unaligned_reads(
unaligned_reads_store,
cfg.rapsearch2_output_file_without_header_coverage,
alignments_test)
# remove temp file
utils.remove_temp_file(unaligned_file_fasta)
# reset the coverage threshold
config.translated_subject_coverage_threshold = current_coverage_threshold
# check the values are unchanged
self.assertEqual(sorted(alignments.get_hit_list()),
sorted(alignments_test.get_hit_list()))
def test_translated_search_unaligned_reads_identity_threshold(self):
"""
Test the unaligned reads function
Test with a rapsearch output file
Test the identity threshold filtering
Test without the coverage filter
"""
# create a set of alignments
alignments = store.Alignments()
# set the coverage threshold to zero so as to not test with filter on
current_coverage_threshold = config.translated_subject_coverage_threshold
config.translated_subject_coverage_threshold = 0
# load the rapsearch output
file_handle = open(cfg.rapsearch2_output_file_with_header)
original_identity_threshold = config.identity_threshold
# set a new threshold that will select 3 of the 5 alignments
config.identity_threshold = 60.0
for line in file_handle:
if not re.search("^#", line):
data = line.strip().split(config.blast_delimiter)
referenceid = data[config.blast_reference_index]
queryid = data[config.blast_query_index]
identity = float(data[config.blast_identity_index])
alignment_length = float(
data[config.blast_aligned_length_index])
# only store those alignments with identities that meet threshold
if identity > config.identity_threshold:
alignments.add(referenceid, 0, queryid,
identity / 100.0 * alignment_length,
"unclassified", alignment_length)
file_handle.close()
alignments_test = store.Alignments()
unaligned_reads_store = store.Reads()
# load the rapsearch output with the unaligned reads function
unaligned_file_fasta = translated.unaligned_reads(
unaligned_reads_store, cfg.rapsearch2_output_file_with_header,
alignments_test)
# remove temp file
utils.remove_temp_file(unaligned_file_fasta)
# reset the coverage threshold
config.translated_subject_coverage_threshold = current_coverage_threshold
# set the threshold back to the default
config.identity_threshold = original_identity_threshold
# check the total number of alignments is the same
self.assertEqual(len(alignments.get_hit_list()),
len(alignments_test.get_hit_list()))