public FullPeptideHit(string line, string sequence, string dynamicModifications, double eValueScore, bool decoy,
Peptide peptide,
IXRawfile2 raw, int scanNumber, int firstScanNumber,
int charge)
: base(sequence, dynamicModifications, eValueScore, decoy, peptide, raw, scanNumber, firstScanNumber, charge)
{
this.line = line;
lineHasExtendedInformation = false;
}
public PeptideHit(string sequence, string dynamicModifications, double eValueScore, bool decoy,
Peptide peptide, IXRawfile2 raw, int scanNumber, int firstScanNumber, int charge)
{
this.sequence = sequence;
this.dynamicModifications = dynamicModifications;
this.eValueScore = eValueScore;
this.decoy = decoy;
this.theoreticalNeutralMass = peptide.Mass;
CalculatePrecursorMassError(raw, scanNumber, firstScanNumber, charge);
this.qValue = double.NaN;
}
public PeptideHit(string sequence, string dynamicModifications, double eValueScore, bool decoy,
Peptide peptide, IXRawfile2 raw, int scanNumber, int firstScanNumber, int charge)
{
this.sequence = sequence;
this.dynamicModifications = dynamicModifications;
this.eValueScore = eValueScore;
this.decoy = decoy;
this.theoreticalNeutralMass = peptide.Mass;
CalculatePrecursorMassError(raw, scanNumber, firstScanNumber, charge);
this.qValue = double.NaN;
}
private void frmMain_DragDrop(object sender, DragEventArgs e)
{
string[] filepaths = (string[])e.Data.GetData(DataFormats.FileDrop);
foreach (string filepath in filepaths)
{
if (Path.GetExtension(filepath).Equals(".csv", StringComparison.InvariantCultureIgnoreCase) &&
!lstOmssaCsvFiles.Items.Contains(filepath))
{
lstOmssaCsvFiles.Items.Add(filepath);
UpdateOutputFolder(filepath);
}
else if (Path.GetExtension(filepath).Equals(".xml", StringComparison.InvariantCultureIgnoreCase))
{
modifications.ReadModificationsFromXmlFile(filepath, true);
Peptide.SetModifications(modifications);
UpdateModsListboxes();
}
}
}
private void frmMain_Load(object sender, EventArgs e)
{
modifications = new ModificationDictionary(Path.Combine(Application.StartupPath, "mods.xml"));
Peptide.SetModifications(modifications);
UpdateModsListboxes();
}
public void Optimize()
{
StreamWriter overall_scans_output = null;
StreamWriter overall_target_output = null;
StreamWriter overall_decoy_output = null;
StreamWriter overall_target_unique_output = null;
StreamWriter overall_decoy_unique_output = null;
StreamWriter overall_scans_phospho_output = null;
StreamWriter overall_target_phospho_output = null;
StreamWriter overall_decoy_phospho_output = null;
StreamWriter overall_target_unique_phospho_output = null;
StreamWriter overall_decoy_unique_phospho_output = null;
StreamWriter overall_log = null;
StreamWriter log = null;
StreamWriter summary = null;
IXRawfile2 raw = null;
StreamReader csv = null;
StreamWriter scans_output = null;
StreamWriter scans_phospho_output = null;
StreamWriter target_output = null;
StreamWriter decoy_output = null;
StreamWriter target_phospho_output = null;
StreamWriter decoy_phospho_output = null;
StreamWriter target_unique_output = null;
StreamWriter decoy_unique_output = null;
StreamWriter target_unique_phospho_output = null;
StreamWriter decoy_unique_phospho_output = null;
StreamWriter overall_target_unique_unique_output = null;
StreamWriter overall_decoy_unique_unique_output = null;
StreamWriter overall_target_unique_unique_phospho_output = null;
StreamWriter overall_decoy_unique_unique_phospho_output = null;
try
{
onStarting(new EventArgs());
onUpdateProgress(new ProgressEventArgs(0));
StringBuilder fixed_modifications_sb = new StringBuilder();
foreach(Modification modification in fixedModifications)
{
fixed_modifications_sb.Append(modification.Name + ", ");
}
if(fixed_modifications_sb.Length > 0)
{
fixed_modifications_sb = fixed_modifications_sb.Remove(fixed_modifications_sb.Length - 2, 2);
}
string fixed_modifications = fixed_modifications_sb.ToString();
Dictionary<string, List<string>> raw_csv_filepaths = new Dictionary<string, List<string>>();
long total_bytes = 0;
foreach(string csv_filepath in csvFilepaths)
{
FileInfo f = new FileInfo(csv_filepath);
total_bytes += f.Length;
string truncated_filename = Path.GetFileNameWithoutExtension(csv_filepath);
string[] raw_filepaths = null;
do
{
if(rawFolder != null && rawFolder != string.Empty && Directory.Exists(rawFolder))
{
raw_filepaths = Directory.GetFiles(rawFolder, truncated_filename + ".raw", SearchOption.AllDirectories);
}
else
{
raw_filepaths = Directory.GetFiles(Directory.GetParent(csv_filepath).ToString(), truncated_filename + ".raw", SearchOption.AllDirectories);
}
truncated_filename = truncated_filename.Substring(0, truncated_filename.Length - 1);
if(truncated_filename.Length == 0)
{
throw new Exception("No corresponding .raw file found for " + csv_filepath);
}
} while(raw_filepaths.Length == 0);
if(!raw_csv_filepaths.ContainsKey(raw_filepaths[0]))
{
raw_csv_filepaths.Add(raw_filepaths[0], new List<string>());
}
raw_csv_filepaths[raw_filepaths[0]].Add(csv_filepath);
}
int overall_scans = 0;
int overall_scans_phospho = 0;
int overall_target = 0;
int overall_target_phospho = 0;
int overall_decoy = 0;
int overall_decoy_phospho = 0;
int overall_target_unique = 0;
int overall_decoy_unique = 0;
int overall_target_unique_phospho = 0;
int overall_decoy_unique_phospho = 0;
int overall_target_unique_unique_phospho = 0;
int overall_decoy_unique_unique_phospho = 0;
if(!Directory.Exists(outputFolder))
{
Directory.CreateDirectory(outputFolder);
}
string overall_output_folder = outputFolder;
string overall_phospho_output_folder = Path.Combine(outputFolder, "phospho");
if(overallOutputs && phosphopeptideOutputs)
{
if(!Directory.Exists(overall_phospho_output_folder))
{
Directory.CreateDirectory(overall_phospho_output_folder);
}
}
string log_folder = Path.Combine(outputFolder, "log");
if(!Directory.Exists(log_folder))
{
Directory.CreateDirectory(log_folder);
}
string scans_folder = Path.Combine(outputFolder, "scans");
if(!Directory.Exists(scans_folder))
{
Directory.CreateDirectory(scans_folder);
}
string scans_phospho_folder = Path.Combine(scans_folder, "phospho");
if(phosphopeptideOutputs)
{
if(!Directory.Exists(scans_phospho_folder))
{
Directory.CreateDirectory(scans_phospho_folder);
}
}
string target_decoy_folder = Path.Combine(outputFolder, "target-decoy");
if(!Directory.Exists(target_decoy_folder))
{
Directory.CreateDirectory(target_decoy_folder);
}
string target_decoy_phospho_folder = Path.Combine(target_decoy_folder, "phospho");
if(phosphopeptideOutputs)
{
if(!Directory.Exists(target_decoy_phospho_folder))
{
Directory.CreateDirectory(target_decoy_phospho_folder);
}
}
string unique_folder = Path.Combine(outputFolder, "unique");
if(!Directory.Exists(unique_folder))
{
Directory.CreateDirectory(unique_folder);
}
string unique_phospho_folder = Path.Combine(unique_folder, "phospho");
if(phosphopeptideOutputs)
{
if(!Directory.Exists(unique_phospho_folder))
{
Directory.CreateDirectory(unique_phospho_folder);
}
}
string overall_scans_filepath = Path.Combine(overall_output_folder, "scans.csv");
string overall_scans_phospho_filepath = Path.Combine(overall_phospho_output_folder, "scans_phospho.csv");
string overall_target_filepath = Path.Combine(overall_output_folder, "target.csv");
string overall_decoy_filepath = Path.Combine(overall_output_folder, "decoy.csv");
string overall_target_phospho_filepath = Path.Combine(overall_phospho_output_folder, "target_phospho.csv");
string overall_decoy_phospho_filepath = Path.Combine(overall_phospho_output_folder, "decoy_phospho.csv");
string overall_target_unique_filepath = Path.Combine(overall_output_folder, "target_unique.csv");
string overall_decoy_unique_filepath = Path.Combine(overall_output_folder, "decoy_unique.csv");
string overall_target_unique_phospho_filepath = Path.Combine(overall_phospho_output_folder, "target_unique_phospho.csv");
string overall_decoy_unique_phospho_filepath = Path.Combine(overall_phospho_output_folder, "decoy_unique_phospho.csv");
if(overallOutputs)
{
overall_log = new StreamWriter(Path.Combine(overall_output_folder, "Batch_FDR_Optimizer_log.txt"));
overall_log.AutoFlush = true;
overall_log.WriteLine("Batch FDR Optimizer PARAMETERS");
overall_log.WriteLine("Fixed Modifications: " + fixed_modifications);
overall_log.WriteLine("Maximum Precursor Mass Error (ppm): ±" + maximumPrecursorMassError.ToString());
overall_log.WriteLine("Precursor Mass Error Increment (ppm): " + precursorMassErrorIncrement.ToString());
overall_log.WriteLine("Higher Scores are Better: " + higherScoresAreBetter.ToString());
overall_log.WriteLine("Maximum False Discovery Rate (%): " + maximumFalseDiscoveryRate.ToString());
overall_log.WriteLine("FDR Calculation and Optimization Based on Unique Peptide Sequences: " + unique.ToString());
overall_log.WriteLine();
overall_scans_output = new StreamWriter(overall_scans_filepath);
overall_target_output = new StreamWriter(overall_target_filepath);
overall_decoy_output = new StreamWriter(overall_decoy_filepath);
overall_target_unique_output = new StreamWriter(overall_target_unique_filepath);
overall_decoy_unique_output = new StreamWriter(overall_decoy_unique_filepath);
if(phosphopeptideOutputs)
{
overall_scans_phospho_output = new StreamWriter(overall_scans_phospho_filepath);
overall_target_phospho_output = new StreamWriter(overall_target_phospho_filepath);
overall_decoy_phospho_output = new StreamWriter(overall_decoy_phospho_filepath);
overall_target_unique_phospho_output = new StreamWriter(overall_target_unique_phospho_filepath);
overall_decoy_unique_phospho_output = new StreamWriter(overall_decoy_unique_phospho_filepath);
}
}
Dictionary<string, List<string>> summary_info = new Dictionary<string, List<string>>();
string extended_header_line = null;
bool overall_header_written = false;
Dictionary<string, SortedDictionary<int, PeptideHit>> all_scans_peptides = new Dictionary<string, SortedDictionary<int, PeptideHit>>();
long bytes_read = 0;
foreach(KeyValuePair<string, List<string>> raw_csv_filepath in raw_csv_filepaths)
{
string raw_filepath = raw_csv_filepath.Key;
raw = (IXRawfile3)new MSFileReader_XRawfile();
raw.Open(raw_filepath);
raw.SetCurrentController(0, 1);
int first_scan_number = 1;
raw.GetFirstSpectrumNumber(ref first_scan_number);
foreach(string csv_filepath in raw_csv_filepath.Value)
{
onStartingFile(new FilepathEventArgs(csv_filepath));
string log_filepath = Path.Combine(log_folder, Path.GetFileNameWithoutExtension(csv_filepath) + "_log.txt");
log = new StreamWriter(log_filepath);
log.AutoFlush = true;
log.WriteLine("Batch FDR Optimizer PARAMETERS");
log.WriteLine("Fixed Modifications: " + fixed_modifications);
log.WriteLine("Maximum Precursor Mass Error (ppm): ±" + maximumPrecursorMassError.ToString());
log.WriteLine("Precursor Mass Error Increment (ppm): " + precursorMassErrorIncrement.ToString());
log.WriteLine("Higher Scores are Better: " + higherScoresAreBetter.ToString());
log.WriteLine("Maximum False Discovery Rate (%): " + maximumFalseDiscoveryRate.ToString());
log.WriteLine("FDR Calculation and Optimization Based on Unique Peptide Sequences: " + unique.ToString());
log.WriteLine();
if(overallOutputs)
{
overall_log.WriteLine(raw_filepath);
overall_log.WriteLine();
}
log.WriteLine(raw_filepath);
log.WriteLine();
SortedDictionary<int, FullPeptideHit> scans_full_peptides = new SortedDictionary<int, FullPeptideHit>();
SortedDictionary<int, PeptideHit> scans_peptides = new SortedDictionary<int, PeptideHit>();
if(overallOutputs)
{
overall_log.WriteLine(csv_filepath);
}
log.WriteLine(csv_filepath);
summary_info.Add(csv_filepath, new List<string>());
summary_info[csv_filepath].Add(raw_filepath);
csv = new StreamReader(csv_filepath);
string header_line = csv.ReadLine();
bytes_read += header_line.Length + 2;
extended_header_line = header_line + ", Precursor Isolation m/z, Precursor Isolation Mass (Da), Precursor Theoretical Neutral Mass (Da), Precursor Experimental Neutral Mass (Da), Precursor Mass Error (ppm), Adjusted Precursor Mass Error (ppm), Q-Value (%)";
if(overallOutputs && !overall_header_written)
{
overall_scans_output.WriteLine(extended_header_line);
overall_target_output.WriteLine(extended_header_line);
overall_decoy_output.WriteLine(extended_header_line);
overall_target_unique_output.WriteLine(extended_header_line);
overall_decoy_unique_output.WriteLine(extended_header_line);
if(phosphopeptideOutputs)
{
overall_scans_phospho_output.WriteLine(extended_header_line);
overall_target_phospho_output.WriteLine(extended_header_line);
overall_decoy_phospho_output.WriteLine(extended_header_line);
overall_target_unique_phospho_output.WriteLine(extended_header_line);
overall_decoy_unique_phospho_output.WriteLine(extended_header_line);
}
overall_header_written = true;
}
/*
using (OmssaReader<DefaultOmssaLine> reader = new OmssaReader<DefaultOmssaLine>(csv_filepath))
{
int counter = 0;
foreach (DefaultOmssaLine psm in reader.ReadNextLine())
{
string line = psm.Line;
bytes_read += line.Length + 2;
int scan_number = psm.SpectrumNumber;
string sequence = psm.Peptide.Sequence;
double evalue_score = psm.EValue;
bool decoy = psm.Decoy;
string dynamic_modifications = psm.Mods;
int charge = psm.Charge;
Peptide peptide = new Peptide(sequence, fixedModifications, dynamic_modifications);
if (!scans_full_peptides.ContainsKey(scan_number))
{
scans_full_peptides.Add(scan_number, new FullPeptideHit(line, sequence, dynamic_modifications, evalue_score, decoy, peptide, raw, scan_number, first_scan_number, charge));
scans_peptides.Add(scan_number, new PeptideHit(sequence, dynamic_modifications, evalue_score, decoy, peptide, raw, scan_number, first_scan_number, charge));
if (dynamic_modifications.Contains("phosphorylation"))
{
overall_scans_phospho++;
}
}
else if ((!higherScoresAreBetter && evalue_score < scans_full_peptides[scan_number].EValueScore
|| (evalue_score == scans_full_peptides[scan_number].EValueScore && scans_full_peptides[scan_number].Decoy && !decoy))
|| (higherScoresAreBetter && evalue_score > scans_full_peptides[scan_number].EValueScore
|| (evalue_score == scans_full_peptides[scan_number].EValueScore && scans_full_peptides[scan_number].Decoy && !decoy)))
{
scans_full_peptides[scan_number] = new FullPeptideHit(line, sequence, dynamic_modifications, evalue_score, decoy, peptide, raw, scan_number, first_scan_number, charge);
scans_peptides[scan_number] = new PeptideHit(sequence, dynamic_modifications, evalue_score, decoy, peptide, raw, scan_number, first_scan_number, charge);
}
counter++;
if (counter % 100 == 1)
{
double progress = (double)bytes_read / total_bytes;
onUpdateProgress(new ProgressEventArgs((int)(progress * 100.0)));
}
}
}
*/
int counter = 0;
while (csv.Peek() != -1)
{
string line = csv.ReadLine();
bytes_read += line.Length + 2;
string[] fields = Regex.Split(line, @",(?!(?<=(?:^|,)\s*\x22(?:[^\x22]|\x22\x22|\\\x22)*,)(?:[^\x22]|\x22\x22|\\\x22)*\x22\s*(?:,|$))"); // crazy regex to parse CSV with internal double quotes from http://regexlib.com/REDetails.aspx?regexp_id=621
int scan_number = int.Parse(fields[0]);
string sequence = fields[2];
double evalue_score = double.Parse(fields[3]);
bool decoy = fields[9].Contains("DECOY") || fields[9].Contains("REVERSED");
string dynamic_modifications = fields[10];
int charge = int.Parse(fields[11]);
Peptide peptide = new Peptide(sequence, fixedModifications, dynamic_modifications);
if (!scans_full_peptides.ContainsKey(scan_number))
{
scans_full_peptides.Add(scan_number, new FullPeptideHit(line, sequence, dynamic_modifications, evalue_score, decoy, peptide, raw, scan_number, first_scan_number, charge));
scans_peptides.Add(scan_number, new PeptideHit(sequence, dynamic_modifications, evalue_score, decoy, peptide, raw, scan_number, first_scan_number, charge));
if (dynamic_modifications.Contains("phosphorylation"))
{
overall_scans_phospho++;
}
}
else if ((!higherScoresAreBetter && evalue_score < scans_full_peptides[scan_number].EValueScore
|| (evalue_score == scans_full_peptides[scan_number].EValueScore && scans_full_peptides[scan_number].Decoy && !decoy))
|| (higherScoresAreBetter && evalue_score > scans_full_peptides[scan_number].EValueScore
|| (evalue_score == scans_full_peptides[scan_number].EValueScore && scans_full_peptides[scan_number].Decoy && !decoy)))
{
scans_full_peptides[scan_number] = new FullPeptideHit(line, sequence, dynamic_modifications, evalue_score, decoy, peptide, raw, scan_number, first_scan_number, charge);
scans_peptides[scan_number] = new PeptideHit(sequence, dynamic_modifications, evalue_score, decoy, peptide, raw, scan_number, first_scan_number, charge);
}
counter++;
if (counter % 100 == 1)
{
double progress = (double)bytes_read / total_bytes;
onUpdateProgress(new ProgressEventArgs((int)(progress * 100.0)));
}
}
csv.Close();
overall_scans += scans_full_peptides.Count;
all_scans_peptides.Add(csv_filepath, scans_peptides);
if(overallOutputs)
{
overall_log.WriteLine();
}
log.WriteLine();
List<PeptideHit> peptides = new List<PeptideHit>(scans_peptides.Values);
peptides.Sort(new AscendingPeptideHitEValueScoreComparer(higherScoresAreBetter));
int cumulative_target_peptides = 0;
int cumulative_decoy_peptides = 0;
double best_low_res_evalue_score_threshold = higherScoresAreBetter ? double.PositiveInfinity : double.NegativeInfinity;
double best_low_res_false_discovery_rate = double.NaN;
int best_target_peptides = 0;
int best_decoy_peptides = 0;
int p = 0;
while(p < peptides.Count)
{
PeptideHit peptide = peptides[p];
if(!peptide.Decoy)
{
cumulative_target_peptides++;
}
else
{
cumulative_decoy_peptides++;
}
p++;
if(p < peptides.Count)
{
PeptideHit next_peptide = peptides[p];
if(next_peptide.EValueScore == peptide.EValueScore)
{
continue;
}
}
double false_discovery_rate = (double)cumulative_decoy_peptides / cumulative_target_peptides * 100.0;
if(false_discovery_rate <= MAXIMUM_FDR_FOR_SYSTEMATIC_PRECURSOR_MASS_ERROR)
{
if(cumulative_target_peptides > best_target_peptides
|| (cumulative_target_peptides == best_target_peptides && false_discovery_rate < best_low_res_false_discovery_rate))
{
best_low_res_evalue_score_threshold = peptide.EValueScore;
best_low_res_false_discovery_rate = false_discovery_rate;
best_target_peptides = cumulative_target_peptides;
best_decoy_peptides = cumulative_decoy_peptides;
}
}
}
List<double> precursor_mass_errors = new List<double>(cumulative_target_peptides);
foreach(PeptideHit peptide in peptides)
{
if((!higherScoresAreBetter && peptide.EValueScore > best_low_res_evalue_score_threshold)
|| (higherScoresAreBetter && peptide.EValueScore < best_low_res_evalue_score_threshold))
{
break;
}
if(!peptide.Decoy)
{
precursor_mass_errors.Add(peptide.PrecursorMassError);
}
}
double median_precursor_mass_error = 0.0;
if(precursor_mass_errors.Count > 0)
{
median_precursor_mass_error = CalculateMedian(precursor_mass_errors);
}
foreach(PeptideHit peptide in scans_peptides.Values)
{
peptide.AdjustedPrecursorMassError = peptide.PrecursorMassError - median_precursor_mass_error;
}
foreach(PeptideHit peptide in scans_full_peptides.Values)
{
peptide.AdjustedPrecursorMassError = peptide.PrecursorMassError - median_precursor_mass_error;
}
List<FullPeptideHit> full_peptides = new List<FullPeptideHit>(scans_full_peptides.Values);
full_peptides.Sort(new AscendingFullPeptideHitEValueScoreComparer(higherScoresAreBetter));
int q_cumulative_target_peptides = 0;
int q_cumulative_decoy_peptides = 0;
Dictionary<string, int> q_target_peptides = new Dictionary<string, int>();
Dictionary<string, int> q_decoy_peptides = new Dictionary<string, int>();
int p3 = 0;
while(p3 < peptides.Count)
{
PeptideHit peptide = peptides[p3];
PeptideHit full_peptide = full_peptides[p3];
if(!peptide.Decoy)
{
q_cumulative_target_peptides++;
if(!q_target_peptides.ContainsKey(peptide.Sequence))
{
q_target_peptides.Add(peptide.Sequence, 0);
}
q_target_peptides[peptide.Sequence]++;
}
else
{
q_cumulative_decoy_peptides++;
if(!q_decoy_peptides.ContainsKey(peptide.Sequence))
{
q_decoy_peptides.Add(peptide.Sequence, 0);
}
q_decoy_peptides[peptide.Sequence]++;
}
p3++;
if(p3 < peptides.Count)
{
PeptideHit next_peptide = peptides[p3];
if(next_peptide.EValueScore == peptide.EValueScore)
{
continue;
}
}
int current_target_peptides = unique ? q_target_peptides.Count : q_cumulative_target_peptides;
int current_decoy_peptides = unique ? q_decoy_peptides.Count : q_cumulative_decoy_peptides;
double q_value = (double)current_decoy_peptides / current_target_peptides * 100.0;
peptide.QValue = q_value;
full_peptide.QValue = q_value;
int p4 = p3 - 1 - 1;
while(p4 >= 0)
{
if(double.IsNaN(peptides[p4].QValue))
{
peptides[p4].QValue = q_value;
full_peptides[p4].QValue = q_value;
p4--;
}
else
{
break;
}
}
}
int p5 = peptides.Count - 1 - 1;
while(p5 >= 0)
{
if(peptides[p5].QValue > peptides[p5 + 1].QValue)
{
peptides[p5].QValue = peptides[p5 + 1].QValue;
full_peptides[p5].QValue = full_peptides[p5 + 1].QValue;
}
p5--;
}
summary_info[csv_filepath].Add(best_low_res_evalue_score_threshold.ToString());
summary_info[csv_filepath].Add(best_target_peptides.ToString());
summary_info[csv_filepath].Add(best_decoy_peptides.ToString());
summary_info[csv_filepath].Add(best_low_res_false_discovery_rate.ToString());
summary_info[csv_filepath].Add(median_precursor_mass_error.ToString());
log.WriteLine("Preliminary E-Value Score Threshold: " + best_low_res_evalue_score_threshold.ToString());
log.WriteLine("Preliminary Target Peptides: " + best_target_peptides.ToString());
log.WriteLine("Preliminary Decoy Peptides: " + best_decoy_peptides.ToString());
log.WriteLine("Preliminary FDR (%): " + best_low_res_false_discovery_rate.ToString());
log.WriteLine("Systematic (Median) Precursor Mass Error (ppm): " + median_precursor_mass_error.ToString());
log.WriteLine();
string scans_filepath = Path.Combine(scans_folder, Path.GetFileNameWithoutExtension(csv_filepath) + "_scans.csv");
scans_output = new StreamWriter(scans_filepath);
scans_output.WriteLine(extended_header_line);
string scans_phospho_filepath = Path.Combine(scans_phospho_folder, Path.GetFileNameWithoutExtension(csv_filepath) + "_scans_phospho.csv");
if(phosphopeptideOutputs)
{
scans_phospho_output = new StreamWriter(scans_phospho_filepath);
scans_phospho_output.WriteLine(extended_header_line);
}
int scans_phospho = 0;
foreach(FullPeptideHit peptide in scans_full_peptides.Values)
{
if(overallOutputs)
{
overall_scans_output.WriteLine(peptide.ExtendedLine);
}
scans_output.WriteLine(peptide.ExtendedLine);
if(phosphopeptideOutputs && peptide.DynamicModifications.Contains("phosphorylation"))
{
scans_phospho++;
if(overallOutputs)
{
overall_scans_phospho_output.WriteLine(peptide.ExtendedLine);
}
scans_phospho_output.WriteLine(peptide.ExtendedLine);
}
}
scans_output.Close();
if(phosphopeptideOutputs)
{
scans_phospho_output.Close();
}
log.WriteLine(scans_filepath);
log.WriteLine(scans_full_peptides.Count.ToString() + " MS/MS scans resulted in at least one peptide hit");
summary_info[csv_filepath].Add(scans_full_peptides.Count.ToString());
if(phosphopeptideOutputs)
{
log.WriteLine(scans_phospho.ToString() + " MS/MS scans resulted in at least one phosphopeptide hit");
summary_info[csv_filepath].Add(scans_phospho.ToString());
}
log.WriteLine();
log.Close();
}
raw.Close();
}
List<PeptideHit> all_peptides = new List<PeptideHit>();
foreach(SortedDictionary<int, PeptideHit> kvp in all_scans_peptides.Values)
{
all_peptides.AddRange(kvp.Values);
}
all_peptides.Sort(new AscendingPeptideHitQValueComparer(higherScoresAreBetter));
int best_target = 0;
int best_decoy = 0;
double best_false_discovery_rate = double.NaN;
double best_q_value = double.NegativeInfinity;
double best_evalue_score = higherScoresAreBetter ? double.PositiveInfinity : double.NegativeInfinity;
double best_max_precursor_mass_error = 0.0;
double max_precursor_mass_error = precursorMassErrorIncrement;
while(max_precursor_mass_error <= maximumPrecursorMassError)
{
List<PeptideHit> all_filtered_peptides = new List<PeptideHit>();
foreach(PeptideHit peptide in all_peptides)
{
if(Math.Abs(peptide.AdjustedPrecursorMassError) <= max_precursor_mass_error)
{
all_filtered_peptides.Add(peptide);
}
}
int target_count = 0;
int decoy_count = 0;
Dictionary<string, int> target_unique_count = new Dictionary<string, int>();
Dictionary<string, int> decoy_unique_count = new Dictionary<string, int>();
int p2 = 0;
while(p2 < all_filtered_peptides.Count)
{
PeptideHit peptide = all_filtered_peptides[p2];
if(!peptide.Decoy)
{
target_count++;
if(!target_unique_count.ContainsKey(peptide.Sequence))
{
target_unique_count.Add(peptide.Sequence, 0);
}
target_unique_count[peptide.Sequence]++;
}
else
{
decoy_count++;
if(!decoy_unique_count.ContainsKey(peptide.Sequence))
{
decoy_unique_count.Add(peptide.Sequence, 0);
}
decoy_unique_count[peptide.Sequence]++;
}
p2++;
if(p2 < all_filtered_peptides.Count)
{
PeptideHit next_peptide = all_filtered_peptides[p2];
if(next_peptide.QValue == peptide.QValue && next_peptide.EValueScore == peptide.EValueScore)
{
continue;
}
}
int target = unique ? target_unique_count.Count : target_count;
int decoy = unique ? decoy_unique_count.Count : decoy_count;
double false_discovery_rate = (double)decoy / target * 100.0;
if(false_discovery_rate <= maximumFalseDiscoveryRate)
{
if(target > best_target || (target == best_target && false_discovery_rate < best_false_discovery_rate))
{
best_target = target;
best_decoy = decoy;
best_false_discovery_rate = false_discovery_rate;
best_q_value = peptide.QValue;
best_evalue_score = peptide.EValueScore;
best_max_precursor_mass_error = max_precursor_mass_error;
}
}
}
max_precursor_mass_error += precursorMassErrorIncrement;
}
SortedDictionary<string, FullPeptideHit> overall_target_peptides = new SortedDictionary<string, FullPeptideHit>();
SortedDictionary<string, FullPeptideHit> overall_decoy_peptides = new SortedDictionary<string, FullPeptideHit>();
foreach(List<string> csv_filepaths in raw_csv_filepaths.Values)
{
foreach(string csv_filepath in csv_filepaths)
{
string log_filepath = Path.Combine(Path.Combine(outputFolder, "log"), Path.GetFileNameWithoutExtension(csv_filepath) + "_log.txt");
log = new StreamWriter(log_filepath, true);
log.AutoFlush = true;
StreamReader scans_csv = new StreamReader(Path.Combine(Path.Combine(outputFolder, "scans"), Path.GetFileNameWithoutExtension(csv_filepath) + "_scans.csv"));
string target_filepath = Path.Combine(target_decoy_folder, Path.GetFileNameWithoutExtension(csv_filepath) + "_target.csv");
target_output = new StreamWriter(target_filepath);
target_output.WriteLine(extended_header_line);
string decoy_filepath = Path.Combine(target_decoy_folder, Path.GetFileNameWithoutExtension(csv_filepath) + "_decoy.csv");
decoy_output = new StreamWriter(decoy_filepath);
decoy_output.WriteLine(extended_header_line);
string target_phospho_filepath = Path.Combine(target_decoy_phospho_folder, Path.GetFileNameWithoutExtension(csv_filepath) + "_target_phospho.csv");
string decoy_phospho_filepath = Path.Combine(target_decoy_phospho_folder, Path.GetFileNameWithoutExtension(csv_filepath) + "_decoy_phospho.csv");
if(phosphopeptideOutputs)
{
target_phospho_output = new StreamWriter(target_phospho_filepath);
target_phospho_output.WriteLine(extended_header_line);
decoy_phospho_output = new StreamWriter(decoy_phospho_filepath);
decoy_phospho_output.WriteLine(extended_header_line);
}
SortedDictionary<string, FullPeptideHit> target_unique = new SortedDictionary<string, FullPeptideHit>();
SortedDictionary<string, FullPeptideHit> decoy_unique = new SortedDictionary<string, FullPeptideHit>();
int redundant_target = 0;
int redundant_decoy = 0;
int redundant_target_phospho = 0;
int redundant_decoy_phospho = 0;
scans_csv.ReadLine();
while(scans_csv.Peek() != -1)
{
string csv_line = scans_csv.ReadLine();
string[] csv_fields = Regex.Split(csv_line, @",(?!(?<=(?:^|,)\s*\x22(?:[^\x22]|\x22\x22|\\\x22)*,)(?:[^\x22]|\x22\x22|\\\x22)*\x22\s*(?:,|$))"); // crazy regex to parse CSV with internal double quotes from http://regexlib.com/REDetails.aspx?regexp_id=621
string sequence = csv_fields[2];
double evalue_score = double.Parse(csv_fields[3]);
bool decoy = csv_fields[9].Contains("DECOY") || csv_fields[9].Contains("REVERSED");
string dynamic_modifications = csv_fields[10];
double isolation_mz = double.Parse(csv_fields[15]);
double isolation_mass = double.Parse(csv_fields[16]);
double theoretical_neutral_mass = double.Parse(csv_fields[17]);
double experimental_neutral_mass = double.Parse(csv_fields[18]);
double precursor_mass_error = double.Parse(csv_fields[19]);
double adjusted_precursor_mass_error = double.Parse(csv_fields[20]);
double q_value = double.Parse(csv_fields[21]);
FullPeptideHit peptide = new FullPeptideHit(csv_line, sequence, dynamic_modifications, evalue_score, decoy, isolation_mz, isolation_mass, theoretical_neutral_mass, experimental_neutral_mass, precursor_mass_error, adjusted_precursor_mass_error, q_value);
if((peptide.QValue < best_q_value || (peptide.QValue == best_q_value
&& ((!higherScoresAreBetter && peptide.EValueScore <= best_evalue_score)
|| (higherScoresAreBetter && peptide.EValueScore >= best_evalue_score))))
&& Math.Abs(peptide.AdjustedPrecursorMassError) <= best_max_precursor_mass_error)
{
if(!peptide.Decoy)
{
redundant_target++;
if(overallOutputs)
{
overall_target_output.WriteLine(peptide.Line);
}
target_output.WriteLine(peptide.Line);
if(phosphopeptideOutputs && peptide.DynamicModifications.Contains("phosphorylation"))
{
redundant_target_phospho++;
if(overallOutputs)
{
overall_target_phospho_output.WriteLine(peptide.Line);
}
target_phospho_output.WriteLine(peptide.Line);
}
if(!target_unique.ContainsKey(peptide.Sequence))
{
target_unique.Add(peptide.Sequence, peptide);
}
else if(peptide.QValue < target_unique[peptide.Sequence].QValue
|| (peptide.QValue == target_unique[peptide.Sequence].QValue
&& ((!higherScoresAreBetter && peptide.EValueScore < target_unique[peptide.Sequence].EValueScore)
|| (higherScoresAreBetter && peptide.EValueScore > target_unique[peptide.Sequence].EValueScore)))
|| (peptide.QValue == target_unique[peptide.Sequence].QValue
&& peptide.EValueScore == target_unique[peptide.Sequence].EValueScore
&& Math.Abs(peptide.AdjustedPrecursorMassError) < Math.Abs(target_unique[peptide.Sequence].AdjustedPrecursorMassError)))
{
target_unique[peptide.Sequence] = peptide;
}
}
else
{
redundant_decoy++;
if(overallOutputs)
{
overall_decoy_output.WriteLine(peptide.Line);
}
decoy_output.WriteLine(peptide.Line);
if(phosphopeptideOutputs && peptide.DynamicModifications.Contains("phosphorylation"))
{
redundant_decoy_phospho++;
if(overallOutputs)
{
overall_decoy_phospho_output.WriteLine(peptide.Line);
}
decoy_phospho_output.WriteLine(peptide.Line);
}
if(!decoy_unique.ContainsKey(peptide.Sequence))
{
decoy_unique.Add(peptide.Sequence, peptide);
}
else if(peptide.QValue < decoy_unique[peptide.Sequence].QValue
|| (peptide.QValue == decoy_unique[peptide.Sequence].QValue
&& ((!higherScoresAreBetter && peptide.EValueScore < decoy_unique[peptide.Sequence].EValueScore)
|| (higherScoresAreBetter && peptide.EValueScore > decoy_unique[peptide.Sequence].EValueScore)))
|| (peptide.QValue == decoy_unique[peptide.Sequence].QValue
&& peptide.EValueScore == decoy_unique[peptide.Sequence].EValueScore
&& Math.Abs(peptide.AdjustedPrecursorMassError) < Math.Abs(decoy_unique[peptide.Sequence].AdjustedPrecursorMassError)))
{
decoy_unique[peptide.Sequence] = peptide;
}
}
}
}
scans_csv.Close();
overall_target += redundant_target;
overall_decoy += redundant_decoy;
overall_target_phospho += redundant_target_phospho;
overall_decoy_phospho += redundant_decoy_phospho;
target_output.Close();
decoy_output.Close();
if(phosphopeptideOutputs)
{
target_phospho_output.Close();
decoy_phospho_output.Close();
}
log.WriteLine("Q-Value Threshold (%): " + best_q_value.ToString());
summary_info[csv_filepath].Add(best_q_value.ToString());
log.WriteLine("E-Value Score Threshold: " + best_evalue_score.ToString());
summary_info[csv_filepath].Add(best_evalue_score.ToString());
log.WriteLine("Maximum Precursor Mass Error (ppm): ±" + best_max_precursor_mass_error.ToString());
summary_info[csv_filepath].Add(best_max_precursor_mass_error.ToString());
log.WriteLine();
log.WriteLine(target_filepath);
log.WriteLine(redundant_target.ToString() + " target peptides after FDR optimization");
summary_info[csv_filepath].Add(redundant_target.ToString());
log.WriteLine(decoy_filepath);
log.WriteLine(redundant_decoy.ToString() + " decoy peptides after FDR optimization");
summary_info[csv_filepath].Add(redundant_decoy.ToString());
if(phosphopeptideOutputs)
{
log.WriteLine(target_phospho_filepath);
log.WriteLine(redundant_target_phospho.ToString() + " target phosphopeptides after FDR optimization");
summary_info[csv_filepath].Add(redundant_target_phospho.ToString());
log.WriteLine(decoy_phospho_filepath);
log.WriteLine(redundant_decoy_phospho.ToString() + " decoy phosphopeptides after FDR optimization");
summary_info[csv_filepath].Add(redundant_decoy_phospho.ToString());
}
log.WriteLine();
if(!unique)
{
log.WriteLine("FDR (%): " + ((double)redundant_decoy / redundant_target * 100.0).ToString());
summary_info[csv_filepath].Add(((double)redundant_decoy / redundant_target * 100.0).ToString());
log.WriteLine();
}
string target_unique_filepath = Path.Combine(unique_folder, Path.GetFileNameWithoutExtension(csv_filepath) + "_target_unique.csv");
target_unique_output = new StreamWriter(target_unique_filepath);
target_unique_output.WriteLine(extended_header_line);
string decoy_unique_filepath = Path.Combine(unique_folder, Path.GetFileNameWithoutExtension(csv_filepath) + "_decoy_unique.csv");
decoy_unique_output = new StreamWriter(decoy_unique_filepath);
decoy_unique_output.WriteLine(extended_header_line);
string target_unique_phospho_filepath = Path.Combine(unique_phospho_folder, Path.GetFileNameWithoutExtension(csv_filepath) + "_target_unique_phospho.csv");
string decoy_unique_phospho_filepath = Path.Combine(unique_phospho_folder, Path.GetFileNameWithoutExtension(csv_filepath) + "_decoy_unique_phospho.csv");
if(phosphopeptideOutputs)
{
target_unique_phospho_output = new StreamWriter(target_unique_phospho_filepath);
target_unique_phospho_output.WriteLine(extended_header_line);
decoy_unique_phospho_output = new StreamWriter(decoy_unique_phospho_filepath);
decoy_unique_phospho_output.WriteLine(extended_header_line);
}
int target_unique_phospho = 0;
foreach(FullPeptideHit peptide in target_unique.Values)
{
if(overallOutputs)
{
overall_target_unique_output.WriteLine(peptide.Line);
}
target_unique_output.WriteLine(peptide.Line);
if(phosphopeptideOutputs && peptide.DynamicModifications.Contains("phosphorylation"))
{
target_unique_phospho++;
if(overallOutputs)
{
overall_target_unique_phospho_output.WriteLine(peptide.Line);
}
target_unique_phospho_output.WriteLine(peptide.Line);
}
if(!overall_target_peptides.ContainsKey(peptide.Sequence))
{
overall_target_peptides.Add(peptide.Sequence, peptide);
}
else if(peptide.QValue < overall_target_peptides[peptide.Sequence].QValue
|| (peptide.QValue == overall_target_peptides[peptide.Sequence].QValue
&& ((!higherScoresAreBetter && peptide.EValueScore < overall_target_peptides[peptide.Sequence].EValueScore)
|| (higherScoresAreBetter && peptide.EValueScore > overall_target_peptides[peptide.Sequence].EValueScore)))
|| (peptide.QValue == overall_target_peptides[peptide.Sequence].QValue
&& peptide.EValueScore == overall_target_peptides[peptide.Sequence].EValueScore
&& Math.Abs(peptide.AdjustedPrecursorMassError) < Math.Abs(overall_target_peptides[peptide.Sequence].AdjustedPrecursorMassError)))
{
overall_target_peptides[peptide.Sequence] = peptide;
}
}
overall_target_unique += target_unique.Count;
overall_target_unique_phospho += target_unique_phospho;
target_unique_output.Close();
if(phosphopeptideOutputs)
{
target_unique_phospho_output.Close();
}
int decoy_unique_phospho = 0;
foreach(FullPeptideHit peptide in decoy_unique.Values)
{
if(overallOutputs)
{
overall_decoy_unique_output.WriteLine(peptide.ExtendedLine);
}
decoy_unique_output.WriteLine(peptide.Line);
if(phosphopeptideOutputs && peptide.DynamicModifications.Contains("phosphorylation"))
{
decoy_unique_phospho++;
if(overallOutputs)
{
overall_decoy_unique_phospho_output.WriteLine(peptide.Line);
}
decoy_unique_phospho_output.WriteLine(peptide.Line);
}
if(!overall_decoy_peptides.ContainsKey(peptide.Sequence))
{
overall_decoy_peptides.Add(peptide.Sequence, peptide);
}
else if(peptide.QValue < overall_decoy_peptides[peptide.Sequence].QValue
|| (peptide.QValue == overall_decoy_peptides[peptide.Sequence].QValue
&& ((!higherScoresAreBetter && peptide.EValueScore < overall_decoy_peptides[peptide.Sequence].EValueScore)
|| (higherScoresAreBetter && peptide.EValueScore > overall_decoy_peptides[peptide.Sequence].EValueScore)))
|| (peptide.QValue == overall_decoy_peptides[peptide.Sequence].QValue
&& peptide.EValueScore == overall_decoy_peptides[peptide.Sequence].EValueScore
&& Math.Abs(peptide.AdjustedPrecursorMassError) < Math.Abs(overall_decoy_peptides[peptide.Sequence].AdjustedPrecursorMassError)))
{
overall_decoy_peptides[peptide.Sequence] = peptide;
}
}
overall_decoy_unique += decoy_unique.Count;
overall_decoy_unique_phospho += decoy_unique_phospho;
decoy_unique_output.Close();
if(phosphopeptideOutputs)
{
decoy_unique_phospho_output.Close();
}
log.WriteLine(target_unique_filepath);
log.WriteLine(target_unique.Count.ToString() + " unique target peptide sequences after FDR optimization");
summary_info[csv_filepath].Add(target_unique.Count.ToString());
log.WriteLine(decoy_unique_filepath);
log.WriteLine(decoy_unique.Count.ToString() + " unique decoy peptide sequences after FDR optimization");
summary_info[csv_filepath].Add(decoy_unique.Count.ToString());
if(phosphopeptideOutputs)
{
log.WriteLine(target_unique_phospho_filepath);
log.WriteLine(target_unique_phospho.ToString() + " unique target phoshopeptide sequences after FDR optimization");
summary_info[csv_filepath].Add(target_unique_phospho.ToString());
log.WriteLine(decoy_unique_phospho_filepath);
log.WriteLine(decoy_unique_phospho.ToString() + " unique decoy phoshopeptide sequences after FDR optimization");
summary_info[csv_filepath].Add(decoy_unique_phospho.ToString());
}
if(unique)
{
log.WriteLine();
log.WriteLine("FDR (%): " + ((double)decoy_unique.Count / target_unique.Count * 100.0).ToString());
summary_info[csv_filepath].Add(((double)decoy_unique.Count / target_unique.Count * 100.0).ToString());
}
log.Close();
onFinishedFile(new FilepathEventArgs(csv_filepath));
}
}
if(overallOutputs)
{
summary_info.Add("SUM", new List<string>());
summary_info["SUM"].Add("n/a");
summary_info["SUM"].Add("n/a");
summary_info["SUM"].Add("n/a");
summary_info["SUM"].Add("n/a");
summary_info["SUM"].Add("n/a");
summary_info["SUM"].Add("n/a");
summary_info.Add("OVERALL", new List<string>());
summary_info["OVERALL"].Add("n/a");
summary_info["OVERALL"].Add("n/a");
summary_info["OVERALL"].Add("n/a");
summary_info["OVERALL"].Add("n/a");
summary_info["OVERALL"].Add("n/a");
summary_info["OVERALL"].Add("n/a");
string overall_target_unique_unique_filepath = Path.Combine(overall_output_folder, "target_unique_unique.csv");
overall_target_unique_unique_output = new StreamWriter(overall_target_unique_unique_filepath);
overall_target_unique_unique_output.WriteLine(extended_header_line);
string overall_decoy_unique_unique_filepath = Path.Combine(overall_output_folder, "decoy_unique_unique.csv");
overall_decoy_unique_unique_output = new StreamWriter(overall_decoy_unique_unique_filepath);
overall_decoy_unique_unique_output.WriteLine(extended_header_line);
string overall_target_unique_unique_phospho_filepath = Path.Combine(overall_phospho_output_folder, "target_unique_unique_phospho.csv");
string overall_decoy_unique_unique_phospho_filepath = Path.Combine(overall_phospho_output_folder, "decoy_unique_unique_phospho.csv");
if(phosphopeptideOutputs)
{
overall_target_unique_unique_phospho_output = new StreamWriter(overall_target_unique_unique_phospho_filepath);
overall_target_unique_unique_phospho_output.WriteLine(extended_header_line);
overall_decoy_unique_unique_phospho_output = new StreamWriter(overall_decoy_unique_unique_phospho_filepath);
overall_decoy_unique_unique_phospho_output.WriteLine(extended_header_line);
}
foreach(FullPeptideHit peptide in overall_target_peptides.Values)
{
overall_target_unique_unique_output.WriteLine(peptide.Line);
if(phosphopeptideOutputs && peptide.DynamicModifications.Contains("phosphorylation"))
{
overall_target_unique_unique_phospho++;
overall_target_unique_unique_phospho_output.WriteLine(peptide.Line);
}
}
foreach(FullPeptideHit peptide in overall_decoy_peptides.Values)
{
overall_decoy_unique_unique_output.WriteLine(peptide.Line);
if(phosphopeptideOutputs && peptide.DynamicModifications.Contains("phosphorylation"))
{
overall_decoy_unique_unique_phospho++;
overall_decoy_unique_unique_phospho_output.WriteLine(peptide.Line);
}
}
overall_scans_output.Close();
overall_target_output.Close();
overall_decoy_output.Close();
overall_target_unique_output.Close();
overall_decoy_unique_output.Close();
overall_target_unique_unique_output.Close();
overall_decoy_unique_unique_output.Close();
if(phosphopeptideOutputs)
{
overall_scans_phospho_output.Close();
overall_target_phospho_output.Close();
overall_decoy_phospho_output.Close();
overall_target_unique_phospho_output.Close();
overall_decoy_unique_phospho_output.Close();
overall_target_unique_unique_phospho_output.Close();
overall_decoy_unique_unique_phospho_output.Close();
}
overall_log.WriteLine(overall_scans_filepath);
overall_log.WriteLine(overall_scans.ToString() + " MS/MS scans resulted in at least one peptide hit");
summary_info["SUM"].Add(overall_scans.ToString());
summary_info["OVERALL"].Add("n/a");
if(phosphopeptideOutputs)
{
overall_log.WriteLine(overall_scans_phospho.ToString() + " MS/MS scans resulted in at least one phosphopeptide hit");
summary_info["SUM"].Add(overall_scans_phospho.ToString());
summary_info["OVERALL"].Add("n/a");
}
overall_log.WriteLine();
overall_log.WriteLine("Q-Value Threshold (%): " + best_q_value.ToString());
summary_info["SUM"].Add(best_q_value.ToString());
summary_info["OVERALL"].Add(best_q_value.ToString());
overall_log.WriteLine("E-Value Score Threshold: " + best_evalue_score.ToString());
summary_info["SUM"].Add(best_evalue_score.ToString());
summary_info["OVERALL"].Add(best_evalue_score.ToString());
overall_log.WriteLine("Maximum Precursor Mass Error (ppm): ±" + best_max_precursor_mass_error.ToString());
summary_info["SUM"].Add(best_max_precursor_mass_error.ToString());
summary_info["OVERALL"].Add(best_max_precursor_mass_error.ToString());
overall_log.WriteLine();
overall_log.WriteLine(overall_target_filepath);
overall_log.WriteLine(overall_target.ToString() + " target peptides after FDR optimization");
summary_info["SUM"].Add(overall_target.ToString());
summary_info["OVERALL"].Add("n/a");
overall_log.WriteLine(overall_decoy_filepath);
overall_log.WriteLine(overall_decoy.ToString() + " decoy peptides after FDR optimization");
summary_info["SUM"].Add(overall_decoy.ToString());
summary_info["OVERALL"].Add("n/a");
if(phosphopeptideOutputs)
{
overall_log.WriteLine(overall_target_phospho_filepath);
overall_log.WriteLine(overall_target_phospho.ToString() + " target phosphopeptides after FDR optimization");
summary_info["SUM"].Add(overall_target_phospho.ToString());
summary_info["OVERALL"].Add("n/a");
overall_log.WriteLine(overall_decoy_phospho_filepath);
overall_log.WriteLine(overall_decoy_phospho.ToString() + " decoy phosphopeptides after FDR optimization");
summary_info["SUM"].Add(overall_decoy_phospho.ToString());
summary_info["OVERALL"].Add("n/a");
}
overall_log.WriteLine();
if(!unique)
{
overall_log.WriteLine("FDR (%): " + ((double)overall_decoy / overall_target * 100.0).ToString());
summary_info["SUM"].Add(((double)overall_decoy / overall_target * 100.0).ToString());
summary_info["OVERALL"].Add("n/a");
overall_log.WriteLine();
}
overall_log.WriteLine(overall_target_unique_filepath);
overall_log.WriteLine(overall_target_unique.ToString() + " target unique peptide sequences after FDR optimization");
summary_info["SUM"].Add(overall_target_unique.ToString());
overall_log.WriteLine(overall_decoy_unique_filepath);
overall_log.WriteLine(overall_decoy_unique.ToString() + " decoy unique peptide sequences after FDR optimization");
summary_info["SUM"].Add(overall_decoy_unique.ToString());
if(phosphopeptideOutputs)
{
overall_log.WriteLine(overall_target_unique_phospho_filepath);
overall_log.WriteLine(overall_target_unique_phospho.ToString() + " target unique phosphopeptide sequences after FDR optimization");
summary_info["SUM"].Add(overall_target_unique_phospho.ToString());
overall_log.WriteLine(overall_decoy_unique_phospho_filepath);
overall_log.WriteLine(overall_decoy_unique_phospho.ToString() + " decoy unique phosphopeptide sequences after FDR optimization");
summary_info["SUM"].Add(overall_decoy_unique_phospho.ToString());
}
overall_log.WriteLine();
overall_log.WriteLine(overall_target_unique_unique_filepath);
overall_log.WriteLine(overall_target_peptides.Count.ToString() + " target unique unique peptide sequences after FDR optimization");
summary_info["OVERALL"].Add(overall_target_peptides.Count.ToString());
overall_log.WriteLine(overall_decoy_unique_unique_filepath);
overall_log.WriteLine(overall_decoy_peptides.Count.ToString() + " decoy unique unique peptide sequences after FDR optimization");
summary_info["OVERALL"].Add(overall_decoy_peptides.Count.ToString());
if(phosphopeptideOutputs)
{
overall_log.WriteLine(overall_target_unique_unique_phospho_filepath);
overall_log.WriteLine(overall_target_unique_unique_phospho.ToString() + " target unique unique phosphopeptide sequences after FDR optimization");
summary_info["OVERALL"].Add(overall_target_unique_unique_phospho.ToString());
overall_log.WriteLine(overall_decoy_unique_unique_phospho_filepath);
overall_log.WriteLine(overall_decoy_unique_unique_phospho.ToString() + " decoy unique unique phosphopeptide sequences after FDR optimization");
summary_info["OVERALL"].Add(overall_decoy_unique_unique_phospho.ToString());
}
if(unique)
{
overall_log.WriteLine();
overall_log.WriteLine("FDR (%): " + ((double)overall_decoy_peptides.Count / overall_target_peptides.Count * 100.0).ToString());
summary_info["SUM"].Add("n/a");
summary_info["OVERALL"].Add(((double)overall_decoy_peptides.Count / overall_target_peptides.Count * 100.0).ToString());
}
overall_log.Close();
summary = new StreamWriter(Path.Combine(overall_output_folder, "summary.csv"));
summary.WriteLine("CSV Filepath,Raw Filepath,Preliminary E-Value Score Threshold,Preliminary Target Peptides,Preliminary Decoy Peptides,Preliminary FDR (%),Systematic (Median) Precursor Mass Error (ppm),Scans," + (phosphopeptideOutputs ? "Phosphopeptide Scans," : null) + "Q-Value Threshold (%),E-Value Score Threshold,Maximum Precursor Mass Error (ppm),Target Peptides,Decoy Peptides," + (phosphopeptideOutputs ? "Target Phosphopeptides,Decoy Phosphopeptides," : null) + (unique ? null : "FDR (%),") + "Unique Target Peptides,Unique Decoy Peptides," + (phosphopeptideOutputs ? "Unique Target Phosphopeptides,Unique Decoy Phosphopeptides," : null) + (unique ? "FDR (%)" : null));
foreach(KeyValuePair<string, List<string>> kvp in summary_info)
{
summary.Write(kvp.Key + ',');
foreach(string item in kvp.Value)
{
summary.Write(item + ',');
}
summary.WriteLine();
}
summary.Close();
}
onFinished(new EventArgs());
}
catch(Exception ex)
{
onThrowException(new ExceptionEventArgs(ex));
}
finally
{
if(overall_scans_output != null)
{
overall_scans_output.Close();
}
if(overall_target_output != null)
{
overall_target_output.Close();
}
if(overall_decoy_output != null)
{
overall_decoy_output.Close();
}
if(overall_target_unique_output != null)
{
overall_target_unique_output.Close();
}
if(overall_decoy_unique_output != null)
{
overall_decoy_unique_output.Close();
}
if(overall_scans_phospho_output != null)
{
overall_scans_phospho_output.Close();
}
if(overall_target_phospho_output != null)
{
overall_target_phospho_output.Close();
}
if(overall_decoy_phospho_output != null)
{
overall_decoy_phospho_output.Close();
}
if(overall_target_unique_phospho_output != null)
{
overall_target_unique_phospho_output.Close();
}
if(overall_decoy_unique_phospho_output != null)
{
overall_decoy_unique_phospho_output.Close();
}
if(overall_log != null)
{
overall_log.Close();
}
if(log != null)
{
log.Close();
}
if(summary != null)
{
summary.Close();
}
if(raw != null)
{
raw.Close();
}
if(csv != null)
{
csv.Close();
}
if(scans_output != null)
{
scans_output.Close();
}
if(target_output != null)
{
target_output.Close();
}
if(decoy_output != null)
{
decoy_output.Close();
}
if(target_phospho_output != null)
{
target_phospho_output.Close();
}
if(decoy_phospho_output != null)
{
decoy_phospho_output.Close();
}
if(target_unique_output != null)
{
target_unique_output.Close();
}
if(decoy_unique_output != null)
{
decoy_unique_output.Close();
}
if(target_unique_phospho_output != null)
{
target_unique_phospho_output.Close();
}
if(decoy_unique_phospho_output != null)
{
decoy_unique_phospho_output.Close();
}
if(overall_target_unique_unique_output != null)
{
overall_target_unique_unique_output.Close();
}
if(overall_decoy_unique_unique_output != null)
{
overall_decoy_unique_unique_output.Close();
}
if(overall_target_unique_unique_phospho_output != null)
{
overall_target_unique_unique_phospho_output.Close();
}
}
}