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Use a Fisher exact test to calculate differential abdunance of each sequence in two samples and reports the log2 transformed fold change, P value and adjusted P value.

Usage

differentialAbundance(
  study_table,
  repertoire_ids = NULL,
  abundance = "duplicate_count",
  type = "junction_aa",
  q = 1,
  zero = 1,
  parallel = FALSE
)

Arguments

study_table

A tibble consisting of antigen receptor sequences imported by the LymphoSeq function readImmunoSeq.

repertoire_ids

A character vector of two repertoire_ids in study_table to be compared. If NULL, the first two repertoire_ids from study_table will be used.

abundance

The input value for the Fisher exact test. "duplicate_count" is recommend but "duplicate_count" may also be used.

type

A character vector indicating whether "junction_aa" or "junction" sequences should be used. If "junction_aa" is specified, then run productiveSeqs first.

q

A numeric value between 0.0 and 1.0 indicating the threshold Holms adjusted P value (also knowns as the false discovery rate or q value) to subset the results with. Any sequences with a q value greater than this value will not be shown.

zero

A numeric value to set all zero values to when calculating the log2 transformed fold change between samples 1 and 2. This does not apply to the p and q value calculations.

parallel

A boolean indicating wheter parallel processing should be used or not.

Value

Returns a data frame with columns corresponding to the frequency of the abudance measure in samples 1 and 2, the P value, Q value (Holms adjusted P value, also knowns as the false discovery rate), and log2 transformed fold change.

Examples

file_path <- system.file("extdata", "TCRB_sequencing", package = "LymphoSeqTest")

stable <- readImmunoSeq(path = file_path)
#> Rows: 1 Columns: 144
#> ── Column specification ────────────────────────────────────────────────────────
#> Delimiter: ","
#> chr (69): sequence_id, sequence, sequence_aa, locus, v_call, d_call, d2_call...
#> dbl (70): v_score, v_identity, v_support, d_score, d_identity, d_support, d2...
#> lgl  (5): rev_comp, productive, vj_in_frame, stop_codon, complete_vdj
#> 
#>  Use `spec()` to retrieve the full column specification for this data.
#>  Specify the column types or set `show_col_types = FALSE` to quiet this message.
#> Rows: 1000 Columns: 52
#> ── Column specification ────────────────────────────────────────────────────────
#> Delimiter: "\t"
#> chr (33): nucleotide, aminoAcid, vMaxResolved, vFamilyName, vGeneName, vGene...
#> dbl (15): count (templates/reads), frequencyCount (%), cdr3Length, vDeletion...
#> lgl  (4): vFamilyTies, jFamilyTies, jGeneNameTies, jGeneAlleleTies
#> 
#>  Use `spec()` to retrieve the full column specification for this data.
#>  Specify the column types or set `show_col_types = FALSE` to quiet this message.
#> Joining, by = c("sequence", "sequence_aa", "v_call", "d_call", "d2_call",
#> "j_call", "junction", "junction_aa", "duplicate_count", "clone_id",
#> "repertoire_id")
#> Rows: 1000 Columns: 52
#> ── Column specification ────────────────────────────────────────────────────────
#> Delimiter: "\t"
#> chr (34): nucleotide, aminoAcid, vMaxResolved, vFamilyName, vGeneName, vGene...
#> dbl (15): count (templates/reads), frequencyCount (%), cdr3Length, vDeletion...
#> lgl  (3): jFamilyTies, jGeneNameTies, jGeneAlleleTies
#> 
#>  Use `spec()` to retrieve the full column specification for this data.
#>  Specify the column types or set `show_col_types = FALSE` to quiet this message.
#> Joining, by = c("sequence", "sequence_aa", "v_call", "d_call", "d2_call",
#> "j_call", "junction", "junction_aa", "duplicate_count", "clone_id",
#> "repertoire_id")
#> Rows: 414 Columns: 52
#> ── Column specification ────────────────────────────────────────────────────────
#> Delimiter: "\t"
#> chr (34): nucleotide, aminoAcid, vMaxResolved, vFamilyName, vGeneName, vGene...
#> dbl (15): count (templates/reads), frequencyCount (%), cdr3Length, vDeletion...
#> lgl  (3): jFamilyTies, jGeneNameTies, jGeneAlleleTies
#> 
#>  Use `spec()` to retrieve the full column specification for this data.
#>  Specify the column types or set `show_col_types = FALSE` to quiet this message.
#> Joining, by = c("sequence", "sequence_aa", "v_call", "d_call", "d2_call",
#> "j_call", "junction", "junction_aa", "duplicate_count", "clone_id",
#> "repertoire_id")
#> Rows: 1000 Columns: 52
#> ── Column specification ────────────────────────────────────────────────────────
#> Delimiter: "\t"
#> chr (34): nucleotide, aminoAcid, vMaxResolved, vFamilyName, vGeneName, vGene...
#> dbl (15): count (templates/reads), frequencyCount (%), cdr3Length, vDeletion...
#> lgl  (3): jFamilyTies, jGeneNameTies, jGeneAlleleTies
#> 
#>  Use `spec()` to retrieve the full column specification for this data.
#>  Specify the column types or set `show_col_types = FALSE` to quiet this message.
#> Joining, by = c("sequence", "sequence_aa", "v_call", "d_call", "d2_call",
#> "j_call", "junction", "junction_aa", "duplicate_count", "clone_id",
#> "repertoire_id")
#> Rows: 1000 Columns: 52
#> ── Column specification ────────────────────────────────────────────────────────
#> Delimiter: "\t"
#> chr (34): nucleotide, aminoAcid, vMaxResolved, vFamilyName, vGeneName, vGene...
#> dbl (15): count (templates/reads), frequencyCount (%), cdr3Length, vDeletion...
#> lgl  (3): jFamilyTies, jGeneNameTies, jGeneAlleleTies
#> 
#>  Use `spec()` to retrieve the full column specification for this data.
#>  Specify the column types or set `show_col_types = FALSE` to quiet this message.
#> Joining, by = c("sequence", "sequence_aa", "v_call", "d_call", "d2_call",
#> "j_call", "junction", "junction_aa", "duplicate_count", "clone_id",
#> "repertoire_id")
#> Rows: 1000 Columns: 52
#> ── Column specification ────────────────────────────────────────────────────────
#> Delimiter: "\t"
#> chr (35): nucleotide, aminoAcid, vMaxResolved, vFamilyName, vGeneName, vGene...
#> dbl (15): count (templates/reads), frequencyCount (%), cdr3Length, vDeletion...
#> lgl  (2): jFamilyTies, jGeneAlleleTies
#> 
#>  Use `spec()` to retrieve the full column specification for this data.
#>  Specify the column types or set `show_col_types = FALSE` to quiet this message.
#> Joining, by = c("sequence", "sequence_aa", "v_call", "d_call", "d2_call",
#> "j_call", "junction", "junction_aa", "duplicate_count", "clone_id",
#> "repertoire_id")
#> Rows: 920 Columns: 52
#> ── Column specification ────────────────────────────────────────────────────────
#> Delimiter: "\t"
#> chr (29): nucleotide, aminoAcid, vMaxResolved, vFamilyName, vGeneName, vFami...
#> dbl (14): count (templates/reads), frequencyCount (%), cdr3Length, vDeletion...
#> lgl  (9): vGeneAllele, vGeneAlleleTies, dGeneAllele, dFamilyTies, dGeneAllel...
#> 
#>  Use `spec()` to retrieve the full column specification for this data.
#>  Specify the column types or set `show_col_types = FALSE` to quiet this message.
#> Joining, by = c("sequence", "sequence_aa", "v_call", "d_call", "d2_call",
#> "j_call", "junction", "junction_aa", "duplicate_count", "clone_id",
#> "repertoire_id")
#> Rows: 1000 Columns: 52
#> ── Column specification ────────────────────────────────────────────────────────
#> Delimiter: "\t"
#> chr (29): nucleotide, aminoAcid, vMaxResolved, vFamilyName, vGeneName, vFami...
#> dbl (14): count (templates/reads), frequencyCount (%), cdr3Length, vDeletion...
#> lgl  (9): vGeneAllele, vGeneAlleleTies, dGeneAllele, dFamilyTies, dGeneAllel...
#> 
#>  Use `spec()` to retrieve the full column specification for this data.
#>  Specify the column types or set `show_col_types = FALSE` to quiet this message.
#> Joining, by = c("sequence", "sequence_aa", "v_call", "d_call", "d2_call",
#> "j_call", "junction", "junction_aa", "duplicate_count", "clone_id",
#> "repertoire_id")
#> Rows: 1000 Columns: 52
#> ── Column specification ────────────────────────────────────────────────────────
#> Delimiter: "\t"
#> chr (29): nucleotide, aminoAcid, vMaxResolved, vFamilyName, vGeneName, vFami...
#> dbl (14): count (templates/reads), frequencyCount (%), cdr3Length, vDeletion...
#> lgl  (9): vGeneAllele, vGeneAlleleTies, dGeneAllele, dFamilyTies, dGeneAllel...
#> 
#>  Use `spec()` to retrieve the full column specification for this data.
#>  Specify the column types or set `show_col_types = FALSE` to quiet this message.
#> Joining, by = c("sequence", "sequence_aa", "v_call", "d_call", "d2_call",
#> "j_call", "junction", "junction_aa", "duplicate_count", "clone_id",
#> "repertoire_id")
#> Rows: 1000 Columns: 52
#> ── Column specification ────────────────────────────────────────────────────────
#> Delimiter: "\t"
#> chr (34): nucleotide, aminoAcid, vMaxResolved, vFamilyName, vGeneName, vGene...
#> dbl (15): count (templates/reads), frequencyCount (%), cdr3Length, vDeletion...
#> lgl  (3): jFamilyTies, jGeneNameTies, jGeneAlleleTies
#> 
#>  Use `spec()` to retrieve the full column specification for this data.
#>  Specify the column types or set `show_col_types = FALSE` to quiet this message.
#> Joining, by = c("sequence", "sequence_aa", "v_call", "d_call", "d2_call",
#> "j_call", "junction", "junction_aa", "duplicate_count", "clone_id",
#> "repertoire_id")

atable <- productiveSeq(study_table = stable, aggregate = "junction_aa")

differentialAbundance(study_table = atable, repertoire_ids = c("TRB_Unsorted_949", "TRB_Unsorted_1320"), 
                      type = "junction_aa", q = 0.01, zero = 0.001)
#> # A tibble: 1,332 × 6
#>    junction_aa       TRB_Unsorted_949 TRB_Unsorted_1320        p       q    l2fc
#>    <chr>                        <dbl>             <dbl>    <dbl>   <dbl>   <dbl>
#>  1 CACALGDGYTF                      0                 2  1   e+0 1   e+0 -Inf   
#>  2 CACQRTGSSYEQYF                   0                 3  1   e+0 1   e+0 -Inf   
#>  3 CAIGLSNQPQHF                     2                67  1   e+0 1   e+0   -5.07
#>  4 CAIKMETPNGEQYF                  29               326  2.66e-5 2.66e-5   -3.49
#>  5 CAIRGTEDNNSPLHF                  0                16  1   e+0 1   e+0 -Inf   
#>  6 CAISDSSYEQYF                     1                23  5.71e-1 5.71e-1   -4.52
#>  7 CAISDTGELFF                      8                83  1.39e-2 1.39e-2   -3.38
#>  8 CAISEFGLMAREYGYTF                0                 1  1   e+0 1   e+0 -Inf   
#>  9 CAISEGQGVKPQHF                   0               167  4.92e-3 4.92e-3 -Inf   
#> 10 CAISESGVLNEKLFF                 13               150  4.69e-3 4.69e-3   -3.53
#> # … with 1,322 more rows