tests/testthat/test_digestFastqs_trans_collapse.R
In fmicompbio/mutscan: Preprocessing and Analysis of Deep Mutational Scanning Data
test_that("digestFastqs works as expected for trans experiments, when similar sequences are collapsed", {
fqt1 <- system.file("extdata/transInput_1.fastq.gz", package = "mutscan")
fqt2 <- system.file("extdata/transInput_2.fastq.gz", package = "mutscan")
## default arguments
Ldef <- list(
fastqForward = fqt1, fastqReverse = fqt2,
mergeForwardReverse = FALSE,
minOverlap = 0, maxOverlap = 0, maxFracMismatchOverlap = 0, greedyOverlap = TRUE,
revComplForward = FALSE, revComplReverse = FALSE,
elementsForward = "SUCV", elementsReverse = "SUCV",
elementLengthsForward = c(1, 10, 18, 96),
elementLengthsReverse = c(1, 8, 20, 96),
adapterForward = "GGAAGAGCACACGTC",
adapterReverse = "GGAAGAGCGTCGTGT",
primerForward = "",
primerReverse = "",
wildTypeForward = "",
wildTypeReverse = "",
constantForward = "AACCGGAGGAGGGAGCTG",
constantReverse = "GAAAAAGGAAGCTGGAGAGA",
avePhredMinForward = 20.0, avePhredMinReverse = 20.0,
variableNMaxForward = 0, variableNMaxReverse = 0,
umiNMax = 0,
nbrMutatedCodonsMaxForward = 1,
nbrMutatedCodonsMaxReverse = 1,
nbrMutatedBasesMaxForward = -1,
nbrMutatedBasesMaxReverse = -1,
forbiddenMutatedCodonsForward = "NNW",
forbiddenMutatedCodonsReverse = "NNW",
useTreeWTmatch = FALSE,
mutatedPhredMinForward = 0.0, mutatedPhredMinReverse = 0.0,
mutNameDelimiter = ".",
constantMaxDistForward = -1,
constantMaxDistReverse = -1,
umiCollapseMaxDist = 4,
filteredReadsFastqForward = "",
filteredReadsFastqReverse = "",
maxNReads = -1, verbose = TRUE,
nThreads = 1, chunkSize = 1000,
maxReadLength = 1024
)
res <- do.call(digestFastqs, Ldef)
## First check that we get deprecation warnings if deprecated arguments are
## specified. Results should be the same as if these arguments are not used
lifecycle::expect_deprecated({
resdep1 <- do.call(digestFastqs, c(Ldef, list(variableCollapseMaxDist = 2)))})
lifecycle::expect_deprecated({
resdep2 <- do.call(digestFastqs, c(Ldef, list(variableCollapseMinReads = 2)))})
lifecycle::expect_deprecated({
resdep3 <- do.call(digestFastqs, c(Ldef, list(variableCollapseMinRatio = 2)))})
expect_equal(res$filterSummary, resdep1$filterSummary)
expect_equal(res$summaryTable, resdep1$summaryTable)
expect_equal(res$errorStatistics, resdep1$errorStatistics)
expect_equal(res$filterSummary, resdep2$filterSummary)
expect_equal(res$summaryTable, resdep2$summaryTable)
expect_equal(res$errorStatistics, resdep2$errorStatistics)
expect_equal(res$filterSummary, resdep3$filterSummary)
expect_equal(res$summaryTable, resdep3$summaryTable)
expect_equal(res$errorStatistics, resdep3$errorStatistics)
## Summarize single sample and collapse
se <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "reads")
secoll <- collapseMutantsBySimilarity(se, assayName = "counts",
collapseMaxDist = 6,
collapseMinScore = 0, collapseMinRatio = 0,
verbose = FALSE)
seumi <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "umis")
secollumi <- collapseMutantsBySimilarity(seumi, assayName = "counts",
scoreMethod = "rowMean",
collapseMaxDist = 6,
collapseMinScore = 0, collapseMinRatio = 0,
verbose = TRUE)
expect_equal(res$filterSummary$nbrTotal, 1000L)
expect_equal(res$filterSummary$f1_nbrAdapter, 314L)
expect_equal(res$filterSummary$f2_nbrNoPrimer, 0L)
expect_equal(res$filterSummary$f3_nbrReadWrongLength, 0L)
expect_equal(res$filterSummary$f4_nbrNoValidOverlap, 0L)
expect_equal(res$filterSummary$f5_nbrAvgVarQualTooLow, 7L)
expect_equal(res$filterSummary$f6_nbrTooManyNinVar, 0L)
expect_equal(res$filterSummary$f7_nbrTooManyNinUMI, 0L)
expect_equal(res$filterSummary$f8_nbrTooManyBestWTHits, 0L)
expect_equal(res$filterSummary$f9_nbrMutQualTooLow, 0L)
expect_equal(res$filterSummary$f10a_nbrTooManyMutCodons, 0L)
expect_equal(res$filterSummary$f10b_nbrTooManyMutBases, 0L)
expect_equal(res$filterSummary$f11_nbrForbiddenCodons, 0L)
expect_equal(res$filterSummary$f12_nbrTooManyMutConstant, 0L)
expect_equal(res$filterSummary$f13_nbrTooManyBestConstantHits, 0L)
expect_equal(res$filterSummary$nbrRetained, 679L)
expect_equal(SummarizedExperiment::colData(secoll)$nbrTotal, 1000L)
expect_equal(SummarizedExperiment::colData(secoll)$f1_nbrAdapter, 314L)
expect_equal(SummarizedExperiment::colData(secoll)$f2_nbrNoPrimer, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f3_nbrReadWrongLength, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f4_nbrNoValidOverlap, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f5_nbrAvgVarQualTooLow, 7L)
expect_equal(SummarizedExperiment::colData(secoll)$f6_nbrTooManyNinVar, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f7_nbrTooManyNinUMI, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f8_nbrTooManyBestWTHits, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f9_nbrMutQualTooLow, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f10a_nbrTooManyMutCodons, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f10b_nbrTooManyMutBases, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f11_nbrForbiddenCodons, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f12_nbrTooManyMutConstant, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f13_nbrTooManyBestConstantHits, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$nbrRetained, 679L)
for (nm in setdiff(names(Ldef), c("forbiddenMutatedCodonsForward", "forbiddenMutatedCodonsReverse", "verbose", "fastqForward", "fastqReverse"))) {
expect_equal(res$parameters[[nm]], Ldef[[nm]], ignore_attr = TRUE)
}
for (nm in c("fastqForward", "fastqReverse")) {
expect_equal(res$parameters[[nm]], normalizePath(Ldef[[nm]], mustWork = FALSE),
ignore_attr = TRUE)
}
expect_equal(sum(res$summaryTable$nbrReads), res$filterSummary$nbrRetained)
expect_equal(sum(SummarizedExperiment::assay(secoll, "counts")),
res$filterSummary$nbrRetained)
expect_equal(nrow(res$summaryTable), 677L)
expect_equal(sum(res$summaryTable$nbrUmis), 679L)
expect_equal(nrow(secoll), 294L)
expect_equal(nrow(secollumi), 294L)
expect_true(all(res$summaryTable$varLengths == "96_96"))
})
test_that("digestFastqs works as expected for trans experiments, when similar sequences are collapsed - extreme case", {
fqt1 <- system.file("extdata/transInput_1.fastq.gz", package = "mutscan")
fqt2 <- system.file("extdata/transInput_2.fastq.gz", package = "mutscan")
## default arguments
Ldef <- list(
fastqForward = fqt1, fastqReverse = fqt2,
mergeForwardReverse = FALSE,
minOverlap = 0, maxOverlap = 0, maxFracMismatchOverlap = 0, greedyOverlap = TRUE,
revComplForward = FALSE, revComplReverse = FALSE,
elementsForward = "SUCV", elementsReverse = "SUCV",
elementLengthsForward = c(1, 10, 18, 96),
elementLengthsReverse = c(1, 8, 20, 96),
adapterForward = "GGAAGAGCACACGTC",
adapterReverse = "GGAAGAGCGTCGTGT",
primerForward = "",
primerReverse = "",
wildTypeForward = "",
wildTypeReverse = "",
constantForward = "AACCGGAGGAGGGAGCTG",
constantReverse = "GAAAAAGGAAGCTGGAGAGA",
avePhredMinForward = 20.0, avePhredMinReverse = 20.0,
variableNMaxForward = 0, variableNMaxReverse = 0,
umiNMax = 0,
nbrMutatedCodonsMaxForward = 1,
nbrMutatedCodonsMaxReverse = 1,
nbrMutatedBasesMaxForward = -1,
nbrMutatedBasesMaxReverse = -1,
forbiddenMutatedCodonsForward = "NNW",
forbiddenMutatedCodonsReverse = "NNW",
useTreeWTmatch = FALSE,
mutatedPhredMinForward = 0.0, mutatedPhredMinReverse = 0.0,
mutNameDelimiter = ".",
constantMaxDistForward = -1,
constantMaxDistReverse = -1,
umiCollapseMaxDist = 100,
filteredReadsFastqForward = "",
filteredReadsFastqReverse = "",
maxNReads = -1, verbose = FALSE,
nThreads = 1, chunkSize = 1000,
maxReadLength = 125
)
res <- do.call(digestFastqs, Ldef)
se <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "reads")
secoll <- collapseMutantsBySimilarity(se, assayName = "counts",
collapseMaxDist = 500,
collapseMinScore = 0, collapseMinRatio = 0,
verbose = FALSE)
seumi <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "umis")
secollumi <- collapseMutantsBySimilarity(seumi, assayName = "counts",
collapseMaxDist = 500,
collapseMinScore = 0, collapseMinRatio = 0,
verbose = FALSE)
expect_equal(res$filterSummary$nbrTotal, 1000L)
expect_equal(res$filterSummary$f1_nbrAdapter, 314L)
expect_equal(res$filterSummary$f2_nbrNoPrimer, 0L)
expect_equal(res$filterSummary$f3_nbrReadWrongLength, 0L)
expect_equal(res$filterSummary$f4_nbrNoValidOverlap, 0L)
expect_equal(res$filterSummary$f5_nbrAvgVarQualTooLow, 7L)
expect_equal(res$filterSummary$f6_nbrTooManyNinVar, 0L)
expect_equal(res$filterSummary$f7_nbrTooManyNinUMI, 0L)
expect_equal(res$filterSummary$f8_nbrTooManyBestWTHits, 0L)
expect_equal(res$filterSummary$f9_nbrMutQualTooLow, 0L)
expect_equal(res$filterSummary$f10a_nbrTooManyMutCodons, 0L)
expect_equal(res$filterSummary$f10b_nbrTooManyMutBases, 0L)
expect_equal(res$filterSummary$f11_nbrForbiddenCodons, 0L)
expect_equal(res$filterSummary$f12_nbrTooManyMutConstant, 0L)
expect_equal(res$filterSummary$f13_nbrTooManyBestConstantHits, 0L)
expect_equal(res$filterSummary$nbrRetained, 679L)
for (nm in setdiff(names(Ldef), c("forbiddenMutatedCodonsForward", "forbiddenMutatedCodonsReverse", "verbose", "fastqForward", "fastqReverse"))) {
expect_equal(res$parameters[[nm]], Ldef[[nm]], ignore_attr = TRUE)
}
for (nm in c("fastqForward", "fastqReverse")) {
expect_equal(res$parameters[[nm]], normalizePath(Ldef[[nm]], mustWork = FALSE),
ignore_attr = TRUE)
}
expect_equal(sum(res$summaryTable$nbrReads), res$filterSummary$nbrRetained)
expect_equal(nrow(res$summaryTable), 677L)
expect_equal(sum(res$summaryTable$nbrUmis), 677L)
expect_equal(nrow(secoll), 1L)
expect_equal(nrow(secollumi), 1L)
})
test_that("digestFastqs works as expected for trans experiments, when similar sequences are collapsed - only forward read", {
fqt1 <- system.file("extdata/transInput_1.fastq.gz", package = "mutscan")
fqt2 <- system.file("extdata/transInput_2.fastq.gz", package = "mutscan")
## default arguments
Ldef <- list(
fastqForward = fqt1, fastqReverse = NULL,
mergeForwardReverse = FALSE,
minOverlap = 0, maxOverlap = 0, maxFracMismatchOverlap = 0, greedyOverlap = TRUE,
revComplForward = FALSE, revComplReverse = FALSE,
elementsForward = "SUCV", elementsReverse = "SUCV",
elementLengthsForward = c(1, 10, 18, 96),
elementLengthsReverse = c(1, 8, 20, 96),
adapterForward = "GGAAGAGCACACGTC",
adapterReverse = "GGAAGAGCGTCGTGT",
primerForward = "",
primerReverse = "",
wildTypeForward = "",
wildTypeReverse = "",
constantForward = "AACCGGAGGAGGGAGCTG",
constantReverse = "GAAAAAGGAAGCTGGAGAGA",
avePhredMinForward = 20.0, avePhredMinReverse = 20.0,
variableNMaxForward = 0, variableNMaxReverse = 0,
umiNMax = 0,
nbrMutatedCodonsMaxForward = 1,
nbrMutatedCodonsMaxReverse = 1,
nbrMutatedBasesMaxForward = -1,
nbrMutatedBasesMaxReverse = -1,
forbiddenMutatedCodonsForward = "NNW",
forbiddenMutatedCodonsReverse = "NNW",
useTreeWTmatch = FALSE,
mutatedPhredMinForward = 0.0, mutatedPhredMinReverse = 0.0,
mutNameDelimiter = ".",
constantMaxDistForward = -1,
constantMaxDistReverse = -1,
umiCollapseMaxDist = 5,
filteredReadsFastqForward = "",
filteredReadsFastqReverse = "",
maxNReads = -1, verbose = FALSE,
nThreads = 1, chunkSize = 1000,
maxReadLength = 1024
)
res <- do.call(digestFastqs, Ldef)
se <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "reads")
secoll <- collapseMutantsBySimilarity(se, assayName = "counts",
collapseMaxDist = 10,
collapseMinScore = 0, collapseMinRatio = 0,
verbose = FALSE)
seumi <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "umis")
secollumi <- collapseMutantsBySimilarity(seumi, assayName = "counts",
collapseMaxDist = 10,
collapseMinScore = 0, collapseMinRatio = 0,
verbose = FALSE)
expect_equal(res$filterSummary$nbrTotal, 1000L)
expect_equal(res$filterSummary$f1_nbrAdapter, 297L)
expect_equal(res$filterSummary$f2_nbrNoPrimer, 0L)
expect_equal(res$filterSummary$f3_nbrReadWrongLength, 0L)
expect_equal(res$filterSummary$f4_nbrNoValidOverlap, 0L)
expect_equal(res$filterSummary$f5_nbrAvgVarQualTooLow, 0L)
expect_equal(res$filterSummary$f6_nbrTooManyNinVar, 0L)
expect_equal(res$filterSummary$f7_nbrTooManyNinUMI, 0L)
expect_equal(res$filterSummary$f8_nbrTooManyBestWTHits, 0L)
expect_equal(res$filterSummary$f9_nbrMutQualTooLow, 0L)
expect_equal(res$filterSummary$f10a_nbrTooManyMutCodons, 0L)
expect_equal(res$filterSummary$f10b_nbrTooManyMutBases, 0L)
expect_equal(res$filterSummary$f11_nbrForbiddenCodons, 0L)
expect_equal(res$filterSummary$f12_nbrTooManyMutConstant, 0L)
expect_equal(res$filterSummary$f13_nbrTooManyBestConstantHits, 0L)
expect_equal(res$filterSummary$nbrRetained, 703L)
for (nm in setdiff(names(Ldef), c("fastqReverse", "forbiddenMutatedCodonsForward", "forbiddenMutatedCodonsReverse", "verbose", "fastqForward"))) {
expect_equal(res$parameters[[nm]], Ldef[[nm]], ignore_attr = TRUE)
}
for (nm in c("fastqForward")) {
expect_equal(res$parameters[[nm]], normalizePath(Ldef[[nm]], mustWork = FALSE),
ignore_attr = TRUE)
}
expect_equal(sum(res$summaryTable$nbrReads), res$filterSummary$nbrRetained)
expect_equal(nrow(res$summaryTable), 613L)
expect_equal(sum(res$summaryTable$nbrUmis), 687L)
expect_equal(nrow(se), 613L)
expect_equal(nrow(secoll), 52L)
expect_equal(nrow(seumi), 613L)
expect_equal(nrow(secollumi), 52L)
})
test_that("digestFastqs works as expected for trans experiments, when similar sequences are collapsed (only abundant ones)", {
fqt1 <- system.file("extdata/transInput_1.fastq.gz", package = "mutscan")
fqt2 <- system.file("extdata/transInput_2.fastq.gz", package = "mutscan")
## default arguments
Ldef <- list(
fastqForward = fqt1, fastqReverse = fqt2,
mergeForwardReverse = FALSE,
minOverlap = 0, maxOverlap = 0, maxFracMismatchOverlap = 0, greedyOverlap = TRUE,
revComplForward = FALSE, revComplReverse = FALSE,
elementsForward = "SUCV", elementsReverse = "SUCV",
elementLengthsForward = c(1, 10, 18, 96),
elementLengthsReverse = c(1, 8, 20, 96),
adapterForward = "GGAAGAGCACACGTC",
adapterReverse = "GGAAGAGCGTCGTGT",
primerForward = "",
primerReverse = "",
wildTypeForward = "",
wildTypeReverse = "",
constantForward = "AACCGGAGGAGGGAGCTG",
constantReverse = "GAAAAAGGAAGCTGGAGAGA",
avePhredMinForward = 20.0, avePhredMinReverse = 20.0,
variableNMaxForward = 0, variableNMaxReverse = 0,
umiNMax = 0,
nbrMutatedCodonsMaxForward = 1,
nbrMutatedCodonsMaxReverse = 1,
nbrMutatedBasesMaxForward = -1,
nbrMutatedBasesMaxReverse = -1,
forbiddenMutatedCodonsForward = "NNW",
forbiddenMutatedCodonsReverse = "NNW",
useTreeWTmatch = FALSE,
mutatedPhredMinForward = 0.0, mutatedPhredMinReverse = 0.0,
mutNameDelimiter = ".",
constantMaxDistForward = -1,
constantMaxDistReverse = -1,
umiCollapseMaxDist = 0,
filteredReadsFastqForward = "",
filteredReadsFastqReverse = "",
maxNReads = -1, verbose = FALSE,
nThreads = 1, chunkSize = 500,
maxReadLength = 1024
)
res <- do.call(digestFastqs, Ldef)
se <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "reads")
secoll <- collapseMutantsBySimilarity(se, assayName = "counts",
collapseMaxDist = 2,
collapseMinScore = 1.5, collapseMinRatio = 0,
verbose = FALSE)
seumi <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "umis")
secollumi <- collapseMutantsBySimilarity(seumi, assayName = "counts",
collapseMaxDist = 2,
collapseMinScore = 1.5, collapseMinRatio = 0,
verbose = FALSE)
expect_equal(res$filterSummary$nbrTotal, 1000L)
expect_equal(res$filterSummary$f1_nbrAdapter, 314L)
expect_equal(res$filterSummary$f2_nbrNoPrimer, 0L)
expect_equal(res$filterSummary$f3_nbrReadWrongLength, 0L)
expect_equal(res$filterSummary$f4_nbrNoValidOverlap, 0L)
expect_equal(res$filterSummary$f5_nbrAvgVarQualTooLow, 7L)
expect_equal(res$filterSummary$f6_nbrTooManyNinVar, 0L)
expect_equal(res$filterSummary$f7_nbrTooManyNinUMI, 0L)
expect_equal(res$filterSummary$f8_nbrTooManyBestWTHits, 0L)
expect_equal(res$filterSummary$f9_nbrMutQualTooLow, 0L)
expect_equal(res$filterSummary$f10a_nbrTooManyMutCodons, 0L)
expect_equal(res$filterSummary$f10b_nbrTooManyMutBases, 0L)
expect_equal(res$filterSummary$f11_nbrForbiddenCodons, 0L)
expect_equal(res$filterSummary$f12_nbrTooManyMutConstant, 0L)
expect_equal(res$filterSummary$f13_nbrTooManyBestConstantHits, 0L)
expect_equal(res$filterSummary$nbrRetained, 679L)
for (nm in setdiff(names(Ldef), c("forbiddenMutatedCodonsForward", "nThreads",
"forbiddenMutatedCodonsReverse", "verbose",
"fastqForward", "fastqReverse"))) {
if (nm == "variableCollapseMinReads") {
expect_equal(res$parameters[[nm]], as.integer(ceiling(Ldef[[nm]])),
ignore_attr = TRUE)
} else {
expect_equal(res$parameters[[nm]], Ldef[[nm]], ignore_attr = TRUE)
}
}
for (nm in c("fastqForward", "fastqReverse")) {
expect_equal(res$parameters[[nm]], normalizePath(Ldef[[nm]], mustWork = FALSE),
ignore_attr = TRUE)
}
expect_equal(sum(res$summaryTable$nbrReads), res$filterSummary$nbrRetained)
expect_equal(nrow(res$summaryTable), 677L)
expect_equal(sum(res$summaryTable$nbrUmis), 679L)
expect_equal(nrow(secoll), 673L)
expect_equal(nrow(secollumi), 673L)
## Don't consider mutations here since we're collapsing (i.e., we have no wildtype)
expect_equal(sum(res$summaryTable$nbrMutBases == 0), nrow(res$summaryTable))
expect_equal(sum(res$summaryTable$nbrMutCodons == 0), nrow(res$summaryTable))
expect_equal(sum(res$summaryTable$nbrMutAAs == 0), nrow(res$summaryTable))
})
test_that("digestFastqs works as expected for trans experiments, when similar sequences are collapsed (only high ratio)", {
fqt1 <- system.file("extdata/transInput_1.fastq.gz", package = "mutscan")
fqt2 <- system.file("extdata/transInput_2.fastq.gz", package = "mutscan")
## default arguments
Ldef <- list(
fastqForward = fqt1, fastqReverse = fqt2,
mergeForwardReverse = FALSE,
minOverlap = 0, maxOverlap = 0, maxFracMismatchOverlap = 0, greedyOverlap = TRUE,
revComplForward = FALSE, revComplReverse = FALSE,
elementsForward = "SUCV", elementsReverse = "SUCV",
elementLengthsForward = c(1, 10, 18, 96),
elementLengthsReverse = c(1, 8, 20, 96),
adapterForward = "GGAAGAGCACACGTC",
adapterReverse = "GGAAGAGCGTCGTGT",
primerForward = "",
primerReverse = "",
wildTypeForward = "",
wildTypeReverse = "",
constantForward = "AACCGGAGGAGGGAGCTG",
constantReverse = "GAAAAAGGAAGCTGGAGAGA",
avePhredMinForward = 20.0, avePhredMinReverse = 20.0,
variableNMaxForward = 0, variableNMaxReverse = 0,
umiNMax = 0,
nbrMutatedCodonsMaxForward = 1,
nbrMutatedCodonsMaxReverse = 1,
nbrMutatedBasesMaxForward = -1,
nbrMutatedBasesMaxReverse = -1,
forbiddenMutatedCodonsForward = "NNW",
forbiddenMutatedCodonsReverse = "NNW",
useTreeWTmatch = FALSE,
mutatedPhredMinForward = 0.0, mutatedPhredMinReverse = 0.0,
mutNameDelimiter = ".",
constantMaxDistForward = -1,
constantMaxDistReverse = -1,
umiCollapseMaxDist = 0,
filteredReadsFastqForward = "",
filteredReadsFastqReverse = "",
maxNReads = -1, verbose = FALSE,
nThreads = 1, chunkSize = 1000,
maxReadLength = 1024
)
res <- do.call(digestFastqs, Ldef)
se <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "reads")
secoll <- collapseMutantsBySimilarity(se, assayName = "counts",
collapseMaxDist = 3,
collapseMinScore = 1, collapseMinRatio = 1.5,
verbose = FALSE)
seumi <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "umis")
secollumi <- collapseMutantsBySimilarity(seumi, assayName = "counts",
collapseMaxDist = 3,
collapseMinScore = 1, collapseMinRatio = 1.5,
verbose = FALSE)
expect_equal(res$filterSummary$nbrTotal, 1000L)
expect_equal(res$filterSummary$f1_nbrAdapter, 314L)
expect_equal(res$filterSummary$f2_nbrNoPrimer, 0L)
expect_equal(res$filterSummary$f3_nbrReadWrongLength, 0L)
expect_equal(res$filterSummary$f4_nbrNoValidOverlap, 0L)
expect_equal(res$filterSummary$f5_nbrAvgVarQualTooLow, 7L)
expect_equal(res$filterSummary$f6_nbrTooManyNinVar, 0L)
expect_equal(res$filterSummary$f7_nbrTooManyNinUMI, 0L)
expect_equal(res$filterSummary$f8_nbrTooManyBestWTHits, 0L)
expect_equal(res$filterSummary$f9_nbrMutQualTooLow, 0L)
expect_equal(res$filterSummary$f10a_nbrTooManyMutCodons, 0L)
expect_equal(res$filterSummary$f10b_nbrTooManyMutBases, 0L)
expect_equal(res$filterSummary$f11_nbrForbiddenCodons, 0L)
expect_equal(res$filterSummary$f12_nbrTooManyMutConstant, 0L)
expect_equal(res$filterSummary$f13_nbrTooManyBestConstantHits, 0L)
expect_equal(res$filterSummary$nbrRetained, 679L)
for (nm in setdiff(names(Ldef), c("forbiddenMutatedCodonsForward", "forbiddenMutatedCodonsReverse", "verbose", "fastqForward", "fastqReverse"))) {
if (nm == "variableCollapseMinReads") {
expect_equal(res$parameters[[nm]], as.integer(ceiling(Ldef[[nm]])),
ignore_attr = TRUE)
} else {
expect_equal(res$parameters[[nm]], Ldef[[nm]], ignore_attr = TRUE)
}
}
for (nm in c("fastqForward", "fastqReverse")) {
expect_equal(res$parameters[[nm]], normalizePath(Ldef[[nm]], mustWork = FALSE),
ignore_attr = TRUE)
}
expect_equal(sum(res$summaryTable$nbrReads), res$filterSummary$nbrRetained)
expect_equal(nrow(res$summaryTable), 677L)
expect_equal(sum(res$summaryTable$nbrUmis), 679L)
expect_equal(nrow(secoll), 656L)
expect_equal(nrow(secollumi), 656L)
})
test_that("digestFastqs works as expected for trans experiments, when similar sequences are collapsed (only high ratio), specify distance threshold as ratio", {
fqt1 <- system.file("extdata/transInput_1.fastq.gz", package = "mutscan")
fqt2 <- system.file("extdata/transInput_2.fastq.gz", package = "mutscan")
## default arguments
Ldef <- list(
fastqForward = fqt1, fastqReverse = fqt2,
mergeForwardReverse = FALSE,
minOverlap = 0, maxOverlap = 0, maxFracMismatchOverlap = 0, greedyOverlap = TRUE,
revComplForward = FALSE, revComplReverse = FALSE,
elementsForward = "SUCV", elementsReverse = "SUCV",
elementLengthsForward = c(1, 10, 18, 96),
elementLengthsReverse = c(1, 8, 20, 96),
adapterForward = "GGAAGAGCACACGTC",
adapterReverse = "GGAAGAGCGTCGTGT",
primerForward = "",
primerReverse = "",
wildTypeForward = "",
wildTypeReverse = "",
constantForward = "AACCGGAGGAGGGAGCTG",
constantReverse = "GAAAAAGGAAGCTGGAGAGA",
avePhredMinForward = 20.0, avePhredMinReverse = 20.0,
variableNMaxForward = 0, variableNMaxReverse = 0,
umiNMax = 0,
nbrMutatedCodonsMaxForward = 1,
nbrMutatedCodonsMaxReverse = 1,
nbrMutatedBasesMaxForward = -1,
nbrMutatedBasesMaxReverse = -1,
forbiddenMutatedCodonsForward = "NNW",
forbiddenMutatedCodonsReverse = "NNW",
useTreeWTmatch = FALSE,
mutatedPhredMinForward = 0.0, mutatedPhredMinReverse = 0.0,
mutNameDelimiter = ".",
constantMaxDistForward = -1,
constantMaxDistReverse = -1,
umiCollapseMaxDist = 0,
filteredReadsFastqForward = "",
filteredReadsFastqReverse = "",
maxNReads = -1, verbose = FALSE,
nThreads = 1, chunkSize = 1000,
maxReadLength = 1024
)
res <- do.call(digestFastqs, Ldef)
se <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "reads")
secoll <- collapseMutantsBySimilarity(se, assayName = "counts",
collapseMaxDist = 0.016,
collapseMinScore = 1, collapseMinRatio = 1.5,
verbose = FALSE)
seumi <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "umis")
secollumi <- collapseMutantsBySimilarity(seumi, assayName = "counts",
collapseMaxDist = 0.016,
collapseMinScore = 1, collapseMinRatio = 1.5,
verbose = FALSE)
expect_equal(res$filterSummary$nbrTotal, 1000L)
expect_equal(res$filterSummary$f1_nbrAdapter, 314L)
expect_equal(res$filterSummary$f2_nbrNoPrimer, 0L)
expect_equal(res$filterSummary$f3_nbrReadWrongLength, 0L)
expect_equal(res$filterSummary$f4_nbrNoValidOverlap, 0L)
expect_equal(res$filterSummary$f5_nbrAvgVarQualTooLow, 7L)
expect_equal(res$filterSummary$f6_nbrTooManyNinVar, 0L)
expect_equal(res$filterSummary$f7_nbrTooManyNinUMI, 0L)
expect_equal(res$filterSummary$f8_nbrTooManyBestWTHits, 0L)
expect_equal(res$filterSummary$f9_nbrMutQualTooLow, 0L)
expect_equal(res$filterSummary$f10a_nbrTooManyMutCodons, 0L)
expect_equal(res$filterSummary$f10b_nbrTooManyMutBases, 0L)
expect_equal(res$filterSummary$f11_nbrForbiddenCodons, 0L)
expect_equal(res$filterSummary$f12_nbrTooManyMutConstant, 0L)
expect_equal(res$filterSummary$f13_nbrTooManyBestConstantHits, 0L)
expect_equal(res$filterSummary$nbrRetained, 679L)
for (nm in setdiff(names(Ldef), c("forbiddenMutatedCodonsForward", "forbiddenMutatedCodonsReverse", "verbose", "fastqForward", "fastqReverse"))) {
if (nm == "variableCollapseMinReads") {
expect_equal(res$parameters[[nm]], as.integer(ceiling(Ldef[[nm]])),
ignore_attr = TRUE)
} else {
expect_equal(res$parameters[[nm]], Ldef[[nm]], ignore_attr = TRUE)
}
}
for (nm in c("fastqForward", "fastqReverse")) {
expect_equal(res$parameters[[nm]], normalizePath(Ldef[[nm]], mustWork = FALSE),
ignore_attr = TRUE)
}
expect_equal(sum(res$summaryTable$nbrReads), res$filterSummary$nbrRetained)
expect_equal(nrow(res$summaryTable), 677L)
expect_equal(sum(res$summaryTable$nbrUmis), 679L)
expect_equal(nrow(secoll), 656L)
expect_equal(nrow(secollumi), 656L)
})
fmicompbio/mutscan documentation built on March 30, 2024, 9:13 a.m.
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test_that("digestFastqs works as expected for trans experiments, when similar sequences are collapsed", {
fqt1 <- system.file("extdata/transInput_1.fastq.gz", package = "mutscan")
fqt2 <- system.file("extdata/transInput_2.fastq.gz", package = "mutscan")
## default arguments
Ldef <- list(
fastqForward = fqt1, fastqReverse = fqt2,
mergeForwardReverse = FALSE,
minOverlap = 0, maxOverlap = 0, maxFracMismatchOverlap = 0, greedyOverlap = TRUE,
revComplForward = FALSE, revComplReverse = FALSE,
elementsForward = "SUCV", elementsReverse = "SUCV",
elementLengthsForward = c(1, 10, 18, 96),
elementLengthsReverse = c(1, 8, 20, 96),
adapterForward = "GGAAGAGCACACGTC",
adapterReverse = "GGAAGAGCGTCGTGT",
primerForward = "",
primerReverse = "",
wildTypeForward = "",
wildTypeReverse = "",
constantForward = "AACCGGAGGAGGGAGCTG",
constantReverse = "GAAAAAGGAAGCTGGAGAGA",
avePhredMinForward = 20.0, avePhredMinReverse = 20.0,
variableNMaxForward = 0, variableNMaxReverse = 0,
umiNMax = 0,
nbrMutatedCodonsMaxForward = 1,
nbrMutatedCodonsMaxReverse = 1,
nbrMutatedBasesMaxForward = -1,
nbrMutatedBasesMaxReverse = -1,
forbiddenMutatedCodonsForward = "NNW",
forbiddenMutatedCodonsReverse = "NNW",
useTreeWTmatch = FALSE,
mutatedPhredMinForward = 0.0, mutatedPhredMinReverse = 0.0,
mutNameDelimiter = ".",
constantMaxDistForward = -1,
constantMaxDistReverse = -1,
umiCollapseMaxDist = 4,
filteredReadsFastqForward = "",
filteredReadsFastqReverse = "",
maxNReads = -1, verbose = TRUE,
nThreads = 1, chunkSize = 1000,
maxReadLength = 1024
)
res <- do.call(digestFastqs, Ldef)
## First check that we get deprecation warnings if deprecated arguments are
## specified. Results should be the same as if these arguments are not used
lifecycle::expect_deprecated({
resdep1 <- do.call(digestFastqs, c(Ldef, list(variableCollapseMaxDist = 2)))})
lifecycle::expect_deprecated({
resdep2 <- do.call(digestFastqs, c(Ldef, list(variableCollapseMinReads = 2)))})
lifecycle::expect_deprecated({
resdep3 <- do.call(digestFastqs, c(Ldef, list(variableCollapseMinRatio = 2)))})
expect_equal(res$filterSummary, resdep1$filterSummary)
expect_equal(res$summaryTable, resdep1$summaryTable)
expect_equal(res$errorStatistics, resdep1$errorStatistics)
expect_equal(res$filterSummary, resdep2$filterSummary)
expect_equal(res$summaryTable, resdep2$summaryTable)
expect_equal(res$errorStatistics, resdep2$errorStatistics)
expect_equal(res$filterSummary, resdep3$filterSummary)
expect_equal(res$summaryTable, resdep3$summaryTable)
expect_equal(res$errorStatistics, resdep3$errorStatistics)
## Summarize single sample and collapse
se <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "reads")
secoll <- collapseMutantsBySimilarity(se, assayName = "counts",
collapseMaxDist = 6,
collapseMinScore = 0, collapseMinRatio = 0,
verbose = FALSE)
seumi <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "umis")
secollumi <- collapseMutantsBySimilarity(seumi, assayName = "counts",
scoreMethod = "rowMean",
collapseMaxDist = 6,
collapseMinScore = 0, collapseMinRatio = 0,
verbose = TRUE)
expect_equal(res$filterSummary$nbrTotal, 1000L)
expect_equal(res$filterSummary$f1_nbrAdapter, 314L)
expect_equal(res$filterSummary$f2_nbrNoPrimer, 0L)
expect_equal(res$filterSummary$f3_nbrReadWrongLength, 0L)
expect_equal(res$filterSummary$f4_nbrNoValidOverlap, 0L)
expect_equal(res$filterSummary$f5_nbrAvgVarQualTooLow, 7L)
expect_equal(res$filterSummary$f6_nbrTooManyNinVar, 0L)
expect_equal(res$filterSummary$f7_nbrTooManyNinUMI, 0L)
expect_equal(res$filterSummary$f8_nbrTooManyBestWTHits, 0L)
expect_equal(res$filterSummary$f9_nbrMutQualTooLow, 0L)
expect_equal(res$filterSummary$f10a_nbrTooManyMutCodons, 0L)
expect_equal(res$filterSummary$f10b_nbrTooManyMutBases, 0L)
expect_equal(res$filterSummary$f11_nbrForbiddenCodons, 0L)
expect_equal(res$filterSummary$f12_nbrTooManyMutConstant, 0L)
expect_equal(res$filterSummary$f13_nbrTooManyBestConstantHits, 0L)
expect_equal(res$filterSummary$nbrRetained, 679L)
expect_equal(SummarizedExperiment::colData(secoll)$nbrTotal, 1000L)
expect_equal(SummarizedExperiment::colData(secoll)$f1_nbrAdapter, 314L)
expect_equal(SummarizedExperiment::colData(secoll)$f2_nbrNoPrimer, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f3_nbrReadWrongLength, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f4_nbrNoValidOverlap, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f5_nbrAvgVarQualTooLow, 7L)
expect_equal(SummarizedExperiment::colData(secoll)$f6_nbrTooManyNinVar, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f7_nbrTooManyNinUMI, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f8_nbrTooManyBestWTHits, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f9_nbrMutQualTooLow, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f10a_nbrTooManyMutCodons, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f10b_nbrTooManyMutBases, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f11_nbrForbiddenCodons, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f12_nbrTooManyMutConstant, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$f13_nbrTooManyBestConstantHits, 0L)
expect_equal(SummarizedExperiment::colData(secoll)$nbrRetained, 679L)
for (nm in setdiff(names(Ldef), c("forbiddenMutatedCodonsForward", "forbiddenMutatedCodonsReverse", "verbose", "fastqForward", "fastqReverse"))) {
expect_equal(res$parameters[[nm]], Ldef[[nm]], ignore_attr = TRUE)
}
for (nm in c("fastqForward", "fastqReverse")) {
expect_equal(res$parameters[[nm]], normalizePath(Ldef[[nm]], mustWork = FALSE),
ignore_attr = TRUE)
}
expect_equal(sum(res$summaryTable$nbrReads), res$filterSummary$nbrRetained)
expect_equal(sum(SummarizedExperiment::assay(secoll, "counts")),
res$filterSummary$nbrRetained)
expect_equal(nrow(res$summaryTable), 677L)
expect_equal(sum(res$summaryTable$nbrUmis), 679L)
expect_equal(nrow(secoll), 294L)
expect_equal(nrow(secollumi), 294L)
expect_true(all(res$summaryTable$varLengths == "96_96"))
})
test_that("digestFastqs works as expected for trans experiments, when similar sequences are collapsed - extreme case", {
fqt1 <- system.file("extdata/transInput_1.fastq.gz", package = "mutscan")
fqt2 <- system.file("extdata/transInput_2.fastq.gz", package = "mutscan")
## default arguments
Ldef <- list(
fastqForward = fqt1, fastqReverse = fqt2,
mergeForwardReverse = FALSE,
minOverlap = 0, maxOverlap = 0, maxFracMismatchOverlap = 0, greedyOverlap = TRUE,
revComplForward = FALSE, revComplReverse = FALSE,
elementsForward = "SUCV", elementsReverse = "SUCV",
elementLengthsForward = c(1, 10, 18, 96),
elementLengthsReverse = c(1, 8, 20, 96),
adapterForward = "GGAAGAGCACACGTC",
adapterReverse = "GGAAGAGCGTCGTGT",
primerForward = "",
primerReverse = "",
wildTypeForward = "",
wildTypeReverse = "",
constantForward = "AACCGGAGGAGGGAGCTG",
constantReverse = "GAAAAAGGAAGCTGGAGAGA",
avePhredMinForward = 20.0, avePhredMinReverse = 20.0,
variableNMaxForward = 0, variableNMaxReverse = 0,
umiNMax = 0,
nbrMutatedCodonsMaxForward = 1,
nbrMutatedCodonsMaxReverse = 1,
nbrMutatedBasesMaxForward = -1,
nbrMutatedBasesMaxReverse = -1,
forbiddenMutatedCodonsForward = "NNW",
forbiddenMutatedCodonsReverse = "NNW",
useTreeWTmatch = FALSE,
mutatedPhredMinForward = 0.0, mutatedPhredMinReverse = 0.0,
mutNameDelimiter = ".",
constantMaxDistForward = -1,
constantMaxDistReverse = -1,
umiCollapseMaxDist = 100,
filteredReadsFastqForward = "",
filteredReadsFastqReverse = "",
maxNReads = -1, verbose = FALSE,
nThreads = 1, chunkSize = 1000,
maxReadLength = 125
)
res <- do.call(digestFastqs, Ldef)
se <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "reads")
secoll <- collapseMutantsBySimilarity(se, assayName = "counts",
collapseMaxDist = 500,
collapseMinScore = 0, collapseMinRatio = 0,
verbose = FALSE)
seumi <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "umis")
secollumi <- collapseMutantsBySimilarity(seumi, assayName = "counts",
collapseMaxDist = 500,
collapseMinScore = 0, collapseMinRatio = 0,
verbose = FALSE)
expect_equal(res$filterSummary$nbrTotal, 1000L)
expect_equal(res$filterSummary$f1_nbrAdapter, 314L)
expect_equal(res$filterSummary$f2_nbrNoPrimer, 0L)
expect_equal(res$filterSummary$f3_nbrReadWrongLength, 0L)
expect_equal(res$filterSummary$f4_nbrNoValidOverlap, 0L)
expect_equal(res$filterSummary$f5_nbrAvgVarQualTooLow, 7L)
expect_equal(res$filterSummary$f6_nbrTooManyNinVar, 0L)
expect_equal(res$filterSummary$f7_nbrTooManyNinUMI, 0L)
expect_equal(res$filterSummary$f8_nbrTooManyBestWTHits, 0L)
expect_equal(res$filterSummary$f9_nbrMutQualTooLow, 0L)
expect_equal(res$filterSummary$f10a_nbrTooManyMutCodons, 0L)
expect_equal(res$filterSummary$f10b_nbrTooManyMutBases, 0L)
expect_equal(res$filterSummary$f11_nbrForbiddenCodons, 0L)
expect_equal(res$filterSummary$f12_nbrTooManyMutConstant, 0L)
expect_equal(res$filterSummary$f13_nbrTooManyBestConstantHits, 0L)
expect_equal(res$filterSummary$nbrRetained, 679L)
for (nm in setdiff(names(Ldef), c("forbiddenMutatedCodonsForward", "forbiddenMutatedCodonsReverse", "verbose", "fastqForward", "fastqReverse"))) {
expect_equal(res$parameters[[nm]], Ldef[[nm]], ignore_attr = TRUE)
}
for (nm in c("fastqForward", "fastqReverse")) {
expect_equal(res$parameters[[nm]], normalizePath(Ldef[[nm]], mustWork = FALSE),
ignore_attr = TRUE)
}
expect_equal(sum(res$summaryTable$nbrReads), res$filterSummary$nbrRetained)
expect_equal(nrow(res$summaryTable), 677L)
expect_equal(sum(res$summaryTable$nbrUmis), 677L)
expect_equal(nrow(secoll), 1L)
expect_equal(nrow(secollumi), 1L)
})
test_that("digestFastqs works as expected for trans experiments, when similar sequences are collapsed - only forward read", {
fqt1 <- system.file("extdata/transInput_1.fastq.gz", package = "mutscan")
fqt2 <- system.file("extdata/transInput_2.fastq.gz", package = "mutscan")
## default arguments
Ldef <- list(
fastqForward = fqt1, fastqReverse = NULL,
mergeForwardReverse = FALSE,
minOverlap = 0, maxOverlap = 0, maxFracMismatchOverlap = 0, greedyOverlap = TRUE,
revComplForward = FALSE, revComplReverse = FALSE,
elementsForward = "SUCV", elementsReverse = "SUCV",
elementLengthsForward = c(1, 10, 18, 96),
elementLengthsReverse = c(1, 8, 20, 96),
adapterForward = "GGAAGAGCACACGTC",
adapterReverse = "GGAAGAGCGTCGTGT",
primerForward = "",
primerReverse = "",
wildTypeForward = "",
wildTypeReverse = "",
constantForward = "AACCGGAGGAGGGAGCTG",
constantReverse = "GAAAAAGGAAGCTGGAGAGA",
avePhredMinForward = 20.0, avePhredMinReverse = 20.0,
variableNMaxForward = 0, variableNMaxReverse = 0,
umiNMax = 0,
nbrMutatedCodonsMaxForward = 1,
nbrMutatedCodonsMaxReverse = 1,
nbrMutatedBasesMaxForward = -1,
nbrMutatedBasesMaxReverse = -1,
forbiddenMutatedCodonsForward = "NNW",
forbiddenMutatedCodonsReverse = "NNW",
useTreeWTmatch = FALSE,
mutatedPhredMinForward = 0.0, mutatedPhredMinReverse = 0.0,
mutNameDelimiter = ".",
constantMaxDistForward = -1,
constantMaxDistReverse = -1,
umiCollapseMaxDist = 5,
filteredReadsFastqForward = "",
filteredReadsFastqReverse = "",
maxNReads = -1, verbose = FALSE,
nThreads = 1, chunkSize = 1000,
maxReadLength = 1024
)
res <- do.call(digestFastqs, Ldef)
se <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "reads")
secoll <- collapseMutantsBySimilarity(se, assayName = "counts",
collapseMaxDist = 10,
collapseMinScore = 0, collapseMinRatio = 0,
verbose = FALSE)
seumi <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "umis")
secollumi <- collapseMutantsBySimilarity(seumi, assayName = "counts",
collapseMaxDist = 10,
collapseMinScore = 0, collapseMinRatio = 0,
verbose = FALSE)
expect_equal(res$filterSummary$nbrTotal, 1000L)
expect_equal(res$filterSummary$f1_nbrAdapter, 297L)
expect_equal(res$filterSummary$f2_nbrNoPrimer, 0L)
expect_equal(res$filterSummary$f3_nbrReadWrongLength, 0L)
expect_equal(res$filterSummary$f4_nbrNoValidOverlap, 0L)
expect_equal(res$filterSummary$f5_nbrAvgVarQualTooLow, 0L)
expect_equal(res$filterSummary$f6_nbrTooManyNinVar, 0L)
expect_equal(res$filterSummary$f7_nbrTooManyNinUMI, 0L)
expect_equal(res$filterSummary$f8_nbrTooManyBestWTHits, 0L)
expect_equal(res$filterSummary$f9_nbrMutQualTooLow, 0L)
expect_equal(res$filterSummary$f10a_nbrTooManyMutCodons, 0L)
expect_equal(res$filterSummary$f10b_nbrTooManyMutBases, 0L)
expect_equal(res$filterSummary$f11_nbrForbiddenCodons, 0L)
expect_equal(res$filterSummary$f12_nbrTooManyMutConstant, 0L)
expect_equal(res$filterSummary$f13_nbrTooManyBestConstantHits, 0L)
expect_equal(res$filterSummary$nbrRetained, 703L)
for (nm in setdiff(names(Ldef), c("fastqReverse", "forbiddenMutatedCodonsForward", "forbiddenMutatedCodonsReverse", "verbose", "fastqForward"))) {
expect_equal(res$parameters[[nm]], Ldef[[nm]], ignore_attr = TRUE)
}
for (nm in c("fastqForward")) {
expect_equal(res$parameters[[nm]], normalizePath(Ldef[[nm]], mustWork = FALSE),
ignore_attr = TRUE)
}
expect_equal(sum(res$summaryTable$nbrReads), res$filterSummary$nbrRetained)
expect_equal(nrow(res$summaryTable), 613L)
expect_equal(sum(res$summaryTable$nbrUmis), 687L)
expect_equal(nrow(se), 613L)
expect_equal(nrow(secoll), 52L)
expect_equal(nrow(seumi), 613L)
expect_equal(nrow(secollumi), 52L)
})
test_that("digestFastqs works as expected for trans experiments, when similar sequences are collapsed (only abundant ones)", {
fqt1 <- system.file("extdata/transInput_1.fastq.gz", package = "mutscan")
fqt2 <- system.file("extdata/transInput_2.fastq.gz", package = "mutscan")
## default arguments
Ldef <- list(
fastqForward = fqt1, fastqReverse = fqt2,
mergeForwardReverse = FALSE,
minOverlap = 0, maxOverlap = 0, maxFracMismatchOverlap = 0, greedyOverlap = TRUE,
revComplForward = FALSE, revComplReverse = FALSE,
elementsForward = "SUCV", elementsReverse = "SUCV",
elementLengthsForward = c(1, 10, 18, 96),
elementLengthsReverse = c(1, 8, 20, 96),
adapterForward = "GGAAGAGCACACGTC",
adapterReverse = "GGAAGAGCGTCGTGT",
primerForward = "",
primerReverse = "",
wildTypeForward = "",
wildTypeReverse = "",
constantForward = "AACCGGAGGAGGGAGCTG",
constantReverse = "GAAAAAGGAAGCTGGAGAGA",
avePhredMinForward = 20.0, avePhredMinReverse = 20.0,
variableNMaxForward = 0, variableNMaxReverse = 0,
umiNMax = 0,
nbrMutatedCodonsMaxForward = 1,
nbrMutatedCodonsMaxReverse = 1,
nbrMutatedBasesMaxForward = -1,
nbrMutatedBasesMaxReverse = -1,
forbiddenMutatedCodonsForward = "NNW",
forbiddenMutatedCodonsReverse = "NNW",
useTreeWTmatch = FALSE,
mutatedPhredMinForward = 0.0, mutatedPhredMinReverse = 0.0,
mutNameDelimiter = ".",
constantMaxDistForward = -1,
constantMaxDistReverse = -1,
umiCollapseMaxDist = 0,
filteredReadsFastqForward = "",
filteredReadsFastqReverse = "",
maxNReads = -1, verbose = FALSE,
nThreads = 1, chunkSize = 500,
maxReadLength = 1024
)
res <- do.call(digestFastqs, Ldef)
se <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "reads")
secoll <- collapseMutantsBySimilarity(se, assayName = "counts",
collapseMaxDist = 2,
collapseMinScore = 1.5, collapseMinRatio = 0,
verbose = FALSE)
seumi <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "umis")
secollumi <- collapseMutantsBySimilarity(seumi, assayName = "counts",
collapseMaxDist = 2,
collapseMinScore = 1.5, collapseMinRatio = 0,
verbose = FALSE)
expect_equal(res$filterSummary$nbrTotal, 1000L)
expect_equal(res$filterSummary$f1_nbrAdapter, 314L)
expect_equal(res$filterSummary$f2_nbrNoPrimer, 0L)
expect_equal(res$filterSummary$f3_nbrReadWrongLength, 0L)
expect_equal(res$filterSummary$f4_nbrNoValidOverlap, 0L)
expect_equal(res$filterSummary$f5_nbrAvgVarQualTooLow, 7L)
expect_equal(res$filterSummary$f6_nbrTooManyNinVar, 0L)
expect_equal(res$filterSummary$f7_nbrTooManyNinUMI, 0L)
expect_equal(res$filterSummary$f8_nbrTooManyBestWTHits, 0L)
expect_equal(res$filterSummary$f9_nbrMutQualTooLow, 0L)
expect_equal(res$filterSummary$f10a_nbrTooManyMutCodons, 0L)
expect_equal(res$filterSummary$f10b_nbrTooManyMutBases, 0L)
expect_equal(res$filterSummary$f11_nbrForbiddenCodons, 0L)
expect_equal(res$filterSummary$f12_nbrTooManyMutConstant, 0L)
expect_equal(res$filterSummary$f13_nbrTooManyBestConstantHits, 0L)
expect_equal(res$filterSummary$nbrRetained, 679L)
for (nm in setdiff(names(Ldef), c("forbiddenMutatedCodonsForward", "nThreads",
"forbiddenMutatedCodonsReverse", "verbose",
"fastqForward", "fastqReverse"))) {
if (nm == "variableCollapseMinReads") {
expect_equal(res$parameters[[nm]], as.integer(ceiling(Ldef[[nm]])),
ignore_attr = TRUE)
} else {
expect_equal(res$parameters[[nm]], Ldef[[nm]], ignore_attr = TRUE)
}
}
for (nm in c("fastqForward", "fastqReverse")) {
expect_equal(res$parameters[[nm]], normalizePath(Ldef[[nm]], mustWork = FALSE),
ignore_attr = TRUE)
}
expect_equal(sum(res$summaryTable$nbrReads), res$filterSummary$nbrRetained)
expect_equal(nrow(res$summaryTable), 677L)
expect_equal(sum(res$summaryTable$nbrUmis), 679L)
expect_equal(nrow(secoll), 673L)
expect_equal(nrow(secollumi), 673L)
## Don't consider mutations here since we're collapsing (i.e., we have no wildtype)
expect_equal(sum(res$summaryTable$nbrMutBases == 0), nrow(res$summaryTable))
expect_equal(sum(res$summaryTable$nbrMutCodons == 0), nrow(res$summaryTable))
expect_equal(sum(res$summaryTable$nbrMutAAs == 0), nrow(res$summaryTable))
})
test_that("digestFastqs works as expected for trans experiments, when similar sequences are collapsed (only high ratio)", {
fqt1 <- system.file("extdata/transInput_1.fastq.gz", package = "mutscan")
fqt2 <- system.file("extdata/transInput_2.fastq.gz", package = "mutscan")
## default arguments
Ldef <- list(
fastqForward = fqt1, fastqReverse = fqt2,
mergeForwardReverse = FALSE,
minOverlap = 0, maxOverlap = 0, maxFracMismatchOverlap = 0, greedyOverlap = TRUE,
revComplForward = FALSE, revComplReverse = FALSE,
elementsForward = "SUCV", elementsReverse = "SUCV",
elementLengthsForward = c(1, 10, 18, 96),
elementLengthsReverse = c(1, 8, 20, 96),
adapterForward = "GGAAGAGCACACGTC",
adapterReverse = "GGAAGAGCGTCGTGT",
primerForward = "",
primerReverse = "",
wildTypeForward = "",
wildTypeReverse = "",
constantForward = "AACCGGAGGAGGGAGCTG",
constantReverse = "GAAAAAGGAAGCTGGAGAGA",
avePhredMinForward = 20.0, avePhredMinReverse = 20.0,
variableNMaxForward = 0, variableNMaxReverse = 0,
umiNMax = 0,
nbrMutatedCodonsMaxForward = 1,
nbrMutatedCodonsMaxReverse = 1,
nbrMutatedBasesMaxForward = -1,
nbrMutatedBasesMaxReverse = -1,
forbiddenMutatedCodonsForward = "NNW",
forbiddenMutatedCodonsReverse = "NNW",
useTreeWTmatch = FALSE,
mutatedPhredMinForward = 0.0, mutatedPhredMinReverse = 0.0,
mutNameDelimiter = ".",
constantMaxDistForward = -1,
constantMaxDistReverse = -1,
umiCollapseMaxDist = 0,
filteredReadsFastqForward = "",
filteredReadsFastqReverse = "",
maxNReads = -1, verbose = FALSE,
nThreads = 1, chunkSize = 1000,
maxReadLength = 1024
)
res <- do.call(digestFastqs, Ldef)
se <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "reads")
secoll <- collapseMutantsBySimilarity(se, assayName = "counts",
collapseMaxDist = 3,
collapseMinScore = 1, collapseMinRatio = 1.5,
verbose = FALSE)
seumi <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "umis")
secollumi <- collapseMutantsBySimilarity(seumi, assayName = "counts",
collapseMaxDist = 3,
collapseMinScore = 1, collapseMinRatio = 1.5,
verbose = FALSE)
expect_equal(res$filterSummary$nbrTotal, 1000L)
expect_equal(res$filterSummary$f1_nbrAdapter, 314L)
expect_equal(res$filterSummary$f2_nbrNoPrimer, 0L)
expect_equal(res$filterSummary$f3_nbrReadWrongLength, 0L)
expect_equal(res$filterSummary$f4_nbrNoValidOverlap, 0L)
expect_equal(res$filterSummary$f5_nbrAvgVarQualTooLow, 7L)
expect_equal(res$filterSummary$f6_nbrTooManyNinVar, 0L)
expect_equal(res$filterSummary$f7_nbrTooManyNinUMI, 0L)
expect_equal(res$filterSummary$f8_nbrTooManyBestWTHits, 0L)
expect_equal(res$filterSummary$f9_nbrMutQualTooLow, 0L)
expect_equal(res$filterSummary$f10a_nbrTooManyMutCodons, 0L)
expect_equal(res$filterSummary$f10b_nbrTooManyMutBases, 0L)
expect_equal(res$filterSummary$f11_nbrForbiddenCodons, 0L)
expect_equal(res$filterSummary$f12_nbrTooManyMutConstant, 0L)
expect_equal(res$filterSummary$f13_nbrTooManyBestConstantHits, 0L)
expect_equal(res$filterSummary$nbrRetained, 679L)
for (nm in setdiff(names(Ldef), c("forbiddenMutatedCodonsForward", "forbiddenMutatedCodonsReverse", "verbose", "fastqForward", "fastqReverse"))) {
if (nm == "variableCollapseMinReads") {
expect_equal(res$parameters[[nm]], as.integer(ceiling(Ldef[[nm]])),
ignore_attr = TRUE)
} else {
expect_equal(res$parameters[[nm]], Ldef[[nm]], ignore_attr = TRUE)
}
}
for (nm in c("fastqForward", "fastqReverse")) {
expect_equal(res$parameters[[nm]], normalizePath(Ldef[[nm]], mustWork = FALSE),
ignore_attr = TRUE)
}
expect_equal(sum(res$summaryTable$nbrReads), res$filterSummary$nbrRetained)
expect_equal(nrow(res$summaryTable), 677L)
expect_equal(sum(res$summaryTable$nbrUmis), 679L)
expect_equal(nrow(secoll), 656L)
expect_equal(nrow(secollumi), 656L)
})
test_that("digestFastqs works as expected for trans experiments, when similar sequences are collapsed (only high ratio), specify distance threshold as ratio", {
fqt1 <- system.file("extdata/transInput_1.fastq.gz", package = "mutscan")
fqt2 <- system.file("extdata/transInput_2.fastq.gz", package = "mutscan")
## default arguments
Ldef <- list(
fastqForward = fqt1, fastqReverse = fqt2,
mergeForwardReverse = FALSE,
minOverlap = 0, maxOverlap = 0, maxFracMismatchOverlap = 0, greedyOverlap = TRUE,
revComplForward = FALSE, revComplReverse = FALSE,
elementsForward = "SUCV", elementsReverse = "SUCV",
elementLengthsForward = c(1, 10, 18, 96),
elementLengthsReverse = c(1, 8, 20, 96),
adapterForward = "GGAAGAGCACACGTC",
adapterReverse = "GGAAGAGCGTCGTGT",
primerForward = "",
primerReverse = "",
wildTypeForward = "",
wildTypeReverse = "",
constantForward = "AACCGGAGGAGGGAGCTG",
constantReverse = "GAAAAAGGAAGCTGGAGAGA",
avePhredMinForward = 20.0, avePhredMinReverse = 20.0,
variableNMaxForward = 0, variableNMaxReverse = 0,
umiNMax = 0,
nbrMutatedCodonsMaxForward = 1,
nbrMutatedCodonsMaxReverse = 1,
nbrMutatedBasesMaxForward = -1,
nbrMutatedBasesMaxReverse = -1,
forbiddenMutatedCodonsForward = "NNW",
forbiddenMutatedCodonsReverse = "NNW",
useTreeWTmatch = FALSE,
mutatedPhredMinForward = 0.0, mutatedPhredMinReverse = 0.0,
mutNameDelimiter = ".",
constantMaxDistForward = -1,
constantMaxDistReverse = -1,
umiCollapseMaxDist = 0,
filteredReadsFastqForward = "",
filteredReadsFastqReverse = "",
maxNReads = -1, verbose = FALSE,
nThreads = 1, chunkSize = 1000,
maxReadLength = 1024
)
res <- do.call(digestFastqs, Ldef)
se <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "reads")
secoll <- collapseMutantsBySimilarity(se, assayName = "counts",
collapseMaxDist = 0.016,
collapseMinScore = 1, collapseMinRatio = 1.5,
verbose = FALSE)
seumi <- summarizeExperiment(list(s1 = res), coldata = data.frame(Name = "s1"),
countType = "umis")
secollumi <- collapseMutantsBySimilarity(seumi, assayName = "counts",
collapseMaxDist = 0.016,
collapseMinScore = 1, collapseMinRatio = 1.5,
verbose = FALSE)
expect_equal(res$filterSummary$nbrTotal, 1000L)
expect_equal(res$filterSummary$f1_nbrAdapter, 314L)
expect_equal(res$filterSummary$f2_nbrNoPrimer, 0L)
expect_equal(res$filterSummary$f3_nbrReadWrongLength, 0L)
expect_equal(res$filterSummary$f4_nbrNoValidOverlap, 0L)
expect_equal(res$filterSummary$f5_nbrAvgVarQualTooLow, 7L)
expect_equal(res$filterSummary$f6_nbrTooManyNinVar, 0L)
expect_equal(res$filterSummary$f7_nbrTooManyNinUMI, 0L)
expect_equal(res$filterSummary$f8_nbrTooManyBestWTHits, 0L)
expect_equal(res$filterSummary$f9_nbrMutQualTooLow, 0L)
expect_equal(res$filterSummary$f10a_nbrTooManyMutCodons, 0L)
expect_equal(res$filterSummary$f10b_nbrTooManyMutBases, 0L)
expect_equal(res$filterSummary$f11_nbrForbiddenCodons, 0L)
expect_equal(res$filterSummary$f12_nbrTooManyMutConstant, 0L)
expect_equal(res$filterSummary$f13_nbrTooManyBestConstantHits, 0L)
expect_equal(res$filterSummary$nbrRetained, 679L)
for (nm in setdiff(names(Ldef), c("forbiddenMutatedCodonsForward", "forbiddenMutatedCodonsReverse", "verbose", "fastqForward", "fastqReverse"))) {
if (nm == "variableCollapseMinReads") {
expect_equal(res$parameters[[nm]], as.integer(ceiling(Ldef[[nm]])),
ignore_attr = TRUE)
} else {
expect_equal(res$parameters[[nm]], Ldef[[nm]], ignore_attr = TRUE)
}
}
for (nm in c("fastqForward", "fastqReverse")) {
expect_equal(res$parameters[[nm]], normalizePath(Ldef[[nm]], mustWork = FALSE),
ignore_attr = TRUE)
}
expect_equal(sum(res$summaryTable$nbrReads), res$filterSummary$nbrRetained)
expect_equal(nrow(res$summaryTable), 677L)
expect_equal(sum(res$summaryTable$nbrUmis), 679L)
expect_equal(nrow(secoll), 656L)
expect_equal(nrow(secollumi), 656L)
})
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