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_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(["s__Bacteroides_xylanisolvens", "unclassified"]))
def test_Alignments_id_mapping_half_hits_with_temp_alignment_file(self):
"""
Test the store_id_mapping function
Test the add_annotated and process_reference_annotation with id mapping
Test the lengths are mapped correctly with only some references included
in those provided for id mapping
Test with the temp alignment file
"""
alignments_store = store.Alignments(minimize_memory_use=True)
# load in the id_mapping file
alignments_store.process_id_mapping(cfg.id_mapping_file)
# store some alignments
alignments_store.add_annotated("query1", 1, "ref1")
alignments_store.add_annotated("query2", 1, "ref2")
alignments_store.add_annotated("query3", 1, "ref1|100")
alignments_store.add_annotated("query3", 1, "200|ref2")
hit_list = alignments_store.get_hit_list()
# delete the temp alignment file
alignments_store.delete_temp_alignments_file()
# test the lengths are correct
stored_lengths = [item[-1] for item in hit_list]
self.assertEqual(
sorted(stored_lengths),
sorted([1 / 1000.0, 100 / 1000.0, 200 / 1000.0, 1000 / 1000.0]))
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()
# 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)
# 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_gene_families_tsv_output_with_names(self):
"""
Test the gene families function and the blast config indexes
Test UniRef50_unknown is read in and used for gene scores but not printed
Test the tsv output
Test that gene families have names applied to them
Test unmapped reads total is written with the same precision as other lines
"""
# update the max decimals to allow for rounding
config.output_max_decimals = 7
# set to a smaller mapping file
original_gene_family_mapping_file = config.gene_family_name_mapping_file
config.gene_family_name_mapping_file = cfg.gene_families_to_names_file
# create a set of alignments
alignments = store.Alignments()
# load the usearch output
file_handle = open(cfg.usearch_uniref50_file)
for line in file_handle:
if not re.search("^#", line):
data = line.strip().split(config.blast_delimiter)
referenceids = data[config.blast_reference_index].split("|")
queryid = data[config.blast_query_index]
identity = float(data[config.blast_identity_index])
alignments.add(referenceids[1], 1, queryid, identity,
referenceids[0])
file_handle.close()
# set the output format
config.output_format = "tsv"
# set the location of the file to write to as a temp file
file_out, gene_families_file = tempfile.mkstemp()
os.close(file_out)
config.genefamilies_file = gene_families_file
# create gene_scores instance
gene_scores = store.GeneScores()
# obtain the gene families
gene_families_file = families.gene_families(alignments, gene_scores, 1)
# check the gene families output is as expected
self.assertTrue(
filecmp.cmp(gene_families_file,
cfg.gene_familes_uniref50_with_names_file,
shallow=False))
# reset the mapping file
config.gene_family_name_mapping_file = original_gene_family_mapping_file
# delete the temp file
utils.remove_temp_file(gene_families_file)
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()
# 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)
# 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 zero as none should pass the threshold)
self.assertEqual(len(alignments.get_hit_list()), 0)
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)
# 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)
# 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_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()
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)
# 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_gene_families_gene_list(self):
"""
Test the gene families function and the blast config indexes
Test UniRef50_unknown is read in and used for gene scores but not printed
Test the gene list
"""
# create a set of alignments
alignments = store.Alignments()
# load the usearch output
file_handle = open(cfg.usearch_file)
for line in file_handle:
if not re.search("^#", line):
data = line.strip().split(config.blast_delimiter)
referenceids = data[config.blast_reference_index].split("|")
queryid = data[config.blast_query_index]
identity = float(data[config.blast_identity_index])
alignments.add(referenceids[1], 1, queryid, identity,
referenceids[0])
file_handle.close()
# check the genes were loaded correctly
self.assertEqual(sorted(cfg.usearch_file_gene_list),
sorted(alignments.gene_list()))
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()
# 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)
# 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_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_nucleotide_search_unaligned_reads_read_count_aligned(self):
"""
Test the unaligned reads and the store alignments
Test with a bowtie2/sam output file
Test for aligned read counts
"""
# create a set of alignments
alignments = store.Alignments()
unaligned_reads_store = store.Reads()
# 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)
# 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)
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()
# 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)
# 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_Alignments_id_mapping_all_bug_list_with_temp_alignment_file(self):
"""
Test the store_id_mapping function
Test the add_annotated and process_reference_annotation with id mapping
Test the bugs are mapped correctly
Test with the temp alignment file
"""
alignments_store = store.Alignments(minimize_memory_use=True)
# load in the id_mapping file
alignments_store.process_id_mapping(cfg.id_mapping_file)
# store some alignments
alignments_store.add_annotated("query1", 1, "ref1")
alignments_store.add_annotated("query2", 1, "ref2")
alignments_store.add_annotated("query3", 1, "ref3")
bug_list = alignments_store.bug_list()
# delete the temp alignment file
alignments_store.delete_temp_alignments_file()
# test the bugs are correct
self.assertEqual(sorted(bug_list), sorted(["bug3", "unclassified"]))
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()
# 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)
# 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_Alignments_process_chocophlan_length(self):
"""
Test the process_chocophlan_length with standard length format
"""
alignments_store=store.Alignments()
length=alignments_store.process_chocophlan_length("1-100","gene")
self.assertEqual(length, 100)
def test_Alignments_process_chocophlan_length_multiple(self):
"""
Test the process_chocophlan_length with multiple lengths
Test with one length on the reverse strand
"""
alignments_store=store.Alignments()
length=alignments_store.process_chocophlan_length("c:100-1,1-100","gene")
self.assertEqual(length, 200)
def test_Alignments_process_reference_annotation_gene_length_reversed(self):
"""
Test the process reference annotation function with a gene and length reversed
"""
alignments_store=store.Alignments()
output=alignments_store.process_reference_annotation("3000|gene")
expected_output=["gene",3000,"unclassified"]
self.assertEqual(expected_output,output)
def test_Alignments_process_reference_annotation_unknown_annotations_three_items_length_string(self):
"""
Test the process reference annotation function with unknown annotations (three items) with string for length
"""
alignments_store=store.Alignments()
output=alignments_store.process_reference_annotation("UniRef90_W1Q3F0|UniRef50_P59787|5000")
expected_output=["UniRef90_W1Q3F0|UniRef50_P59787|5000",0,"unclassified"]
self.assertEqual(expected_output,output)
def test_Alignments_process_reference_annotation_unknown_annotations_four_items(self):
"""
Test the process reference annotation function with unknown annotations (four items)
"""
alignments_store=store.Alignments()
output=alignments_store.process_reference_annotation("UniRef90_W1Q3F0|UniRef50_P59787|5000|bug")
expected_output=["UniRef90_W1Q3F0|UniRef50_P59787|5000|bug",0,"unclassified"]
self.assertEqual(expected_output,output)
def test_Alignments_process_reference_annotation_gene_length_with_bug(self):
"""
Test the process reference annotation function with a gene and length and bug
"""
alignments_store=store.Alignments()
output=alignments_store.process_reference_annotation("gene|3000|bug")
expected_output=["gene",3000,"bug"]
self.assertEqual(expected_output,output)
def test_Alignments_process_reference_annotation_unknown_annotations_three_items_bug_int(self):
"""
Test the process reference annotation function with unknown annotations (three items) with int as bug
"""
alignments_store=store.Alignments()
output=alignments_store.process_reference_annotation("UniRef90_W1Q3F0|5000|5000")
expected_output=["UniRef90_W1Q3F0|5000|5000",0,"unclassified"]
self.assertEqual(expected_output,output)
def test_Alignments_process_reference_annotation_numerical_gene_length(self):
"""
Test the process reference annotation function with gene (as number) and length
"""
alignments_store=store.Alignments()
output=alignments_store.process_reference_annotation("59787|5000")
expected_output=["59787",5000,"unclassified"]
self.assertEqual(expected_output,output)
def test_Alignments_compute_gene_scores_double_gene_double_query_with_temp_alignment_file(
self):
"""
Test the compute_gene_scores function
Test two hits to gene with more than one hit per query
Test with the temp alignment file
"""
# create a set of hits
# bug, reference, reference_length, query, matches = hit
matches1 = 41.0
matches2 = 57.1
matches3 = 61.0
matches4 = 72.1
gene1_length = 2
gene2_length = 3
gene3_length = 4
# Create a set of alignments
alignments_store = store.Alignments(minimize_memory_use=True)
alignments_store.add("gene1", gene1_length, "query1", matches1, "bug1")
alignments_store.add("gene2", gene2_length, "query1", matches2, "bug1")
alignments_store.add("gene2", gene2_length, "query2", matches3, "bug1")
alignments_store.add("gene3", gene3_length, "query3", matches4, "bug1")
gene_scores_store = store.GeneScores()
# compute gene scores
alignments_store.convert_alignments_to_gene_scores(gene_scores_store)
# gene1
hit1_score = math.pow(matches1, config.match_power)
hit2_score = math.pow(matches2, config.match_power)
query1_sum = hit1_score + hit2_score
# convert lengths to per kb
gene2_length = gene2_length / 1000.0
# gene2
hit3_score = math.pow(matches3, config.match_power)
query2_sum = hit3_score
expected_gene_score = hit3_score / query2_sum / gene2_length + hit2_score / query1_sum / gene2_length
actual_gene_score = gene_scores_store.get_score("bug1", "gene2")
# delete the temp alignment file
alignments_store.delete_temp_alignments_file()
self.assertAlmostEqual(actual_gene_score,
expected_gene_score,
places=7)
def test_Alignments_process_reference_annotation_new_chocophlan_annotations(self):
"""
Test the process reference annotation function with the new chocophlan annotations
"""
alignments_store=store.Alignments()
output=alignments_store.process_reference_annotation(
"gi|554771211|gb|ACIN03000006.1|:c1189-5|46125|g__Abiotrophia.s__Abiotrophia_defectiva|UniRef90_W1Q3F0|UniRef50_P59787|5000")
expected_output=["UniRef50_P59787",5000,"g__Abiotrophia.s__Abiotrophia_defectiva"]
self.assertEqual(expected_output,output)
def test_Alignments_add_gene_count(self):
"""
Alignments class: Test add function
Test the total genes
"""
alignments_store=store.Alignments()
alignments_store.add("gene2", 1, "Q3", 0.01, "bug1",1)
alignments_store.add("gene1", 1, "Q1", 0.01, "bug2",1)
alignments_store.add("gene3", 1, "Q2", 0.01, "bug3",1)
alignments_store.add("gene1", 1, "Q1", 0.01, "bug1",1)
# check the total genes
self.assertEqual(alignments_store.count_genes(),3)
def test_Alignments_add_gene_list(self):
"""
Alignments class: Test add function
Test the gene list
"""
alignments_store=store.Alignments()
alignments_store.add("gene2", 1, "Q3", 0.01, "bug1",1)
alignments_store.add("gene1", 1, "Q1", 0.01, "bug2",1)
alignments_store.add("gene3", 1, "Q2", 0.01, "bug3",1)
alignments_store.add("gene1", 1, "Q1", 0.01, "bug1",1)
# check gene list
self.assertEqual(sorted(alignments_store.gene_list()),["gene1","gene2","gene3"])
def test_Alignments_add_gene_lengths(self):
"""
Alignments class: Test add function
Test the gene lengths
"""
alignments_store=store.Alignments()
alignments_store.add("gene2", 10, "Q3", 0.01, "bug1",1)
alignments_store.add("gene1", 100, "Q1", 0.01, "bug2",1)
alignments_store.add("gene3", 1000, "Q2", 0.01, "bug3",1)
alignments_store.add("gene1", 0, "Q1", 0.01, "bug1",1)
# test the lengths are correct
stored_lengths=[item[-1] for item in alignments_store.get_hit_list()]
self.assertEqual(sorted(stored_lengths),sorted([10/1000.0,100/1000.0,1000/1000.0,1000/1000.0]))
def test_Alignments_compute_gene_scores_single_gene_single_query_with_temp_alignment_file(
self):
"""
Test the compute_gene_scores function
Test one hit for gene with one hit for query
Test with the temp alignment file
"""
# create a set of hits
matches1 = 41.0
matches2 = 57.1
matches3 = 61.0
matches4 = 72.1
gene1_length = 2
gene2_length = 3
gene3_length = 4
# Create a set of alignments
alignments_store = store.Alignments(minimize_memory_use=True)
alignments_store.add("gene1", gene1_length, "query1", matches1, "bug1")
alignments_store.add("gene2", gene2_length, "query1", matches2, "bug1")
alignments_store.add("gene2", gene2_length, "query2", matches3, "bug1")
alignments_store.add("gene3", gene3_length, "query3", matches4, "bug1")
gene_scores_store = store.GeneScores()
# compute gene scores
alignments_store.convert_alignments_to_gene_scores(gene_scores_store)
# convert lengths to per kb
gene3_length = gene3_length / 1000.0
# gene3
hit4_score = math.pow(matches4, config.match_power)
query3_sum = hit4_score
expected_gene_score = hit4_score / query3_sum / gene3_length
actual_gene_score = gene_scores_store.get_score("bug1", "gene3")
# delete the temp alignment file
alignments_store.delete_temp_alignments_file()
self.assertEqual(actual_gene_score, expected_gene_score)
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)
