test_diff: Differential enrichment test

Description Usage Arguments Value Examples

View source: R/functions.R

Description

test_diff performs a differential enrichment test based on protein-wise linear models and empirical Bayes statistics using limma. False Discovery Rates are estimated using fdrtool.

Usage

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test_diff(se, type = c("control", "all", "manual"), control = NULL,
  test = NULL, design_formula = formula(~0 + condition))

Arguments

se

SummarizedExperiment, Proteomics data (output from make_se() or make_se_parse()). It is adviced to first remove proteins with too many missing values using filter_missval(), normalize the data using normalize_vsn() and impute remaining missing values using impute().

type

"control", "all" or "manual", The type of contrasts that will be tested. This can be all possible pairwise comparisons ("all"), limited to the comparisons versus the control ("control"), or manually defined contrasts ("manual").

control

Character(1), The condition to which contrasts are generated if type = "control" (a control condition would be most appropriate).

test

Character, The contrasts that will be tested if type = "manual". These should be formatted as "SampleA_vs_SampleB" or c("SampleA_vs_SampleC", "SampleB_vs_SampleC").

design_formula

Formula, Used to create the design matrix.

Value

A SummarizedExperiment object containing fdr estimates of differential expression.

Examples

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# Load example
data <- UbiLength
data <- data[data$Reverse != "+" & data$Potential.contaminant != "+",]
data_unique <- make_unique(data, "Gene.names", "Protein.IDs", delim = ";")

# Make SummarizedExperiment
columns <- grep("LFQ.", colnames(data_unique))
exp_design <- UbiLength_ExpDesign
se <- make_se(data_unique, columns, exp_design)

# Filter, normalize and impute missing values
filt <- filter_missval(se, thr = 0)
norm <- normalize_vsn(filt)
imputed <- impute(norm, fun = "MinProb", q = 0.01)

# Test for differentially expressed proteins
diff <- test_diff(imputed, "control", "Ctrl")
diff <- test_diff(imputed, "manual",
    test = c("Ubi4_vs_Ctrl", "Ubi6_vs_Ctrl"))

# Test for differentially expressed proteins with a custom design formula
diff <- test_diff(imputed, "control", "Ctrl",
    design_formula = formula(~ 0 + condition + replicate))

DEP documentation built on Nov. 8, 2020, 7:49 p.m.