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MikkelSchubert / adapterremoval / #111

30 Apr 2025 09:52AM UTC coverage: 67.045% (+0.06%) from 66.988%
#111

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fix misleading io error messages (#137)

In particular, errors without assosiated error codes would include
nonsensical error descriptions. The use of ios_base::failure is dropped,
since IO generally is not done using iostreams

2 of 8 new or added lines in 4 files covered. (25.0%)

9751 of 14544 relevant lines covered (67.04%)

3041.83 hits per line

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0.0
/src/reports_html.cpp
1
// SPDX-License-Identifier: GPL-3.0-or-later
2
// SPDX-FileCopyrightText: 2022 Mikkel Schubert <mikkelsch@gmail.com>
3
#include "adapter_id.hpp"            // for adapter_id_statistics
4
#include "counts.hpp"                // for counts, indexed_count, counts_tmpl
5
#include "debug.hpp"                 // for AR_REQUIRE
6
#include "errors.hpp"                // for io_error
7
#include "fastq.hpp"                 // for ACGT, ACGT::values, fastq, ACGTN
8
#include "json.hpp"                  // for json_dict, json_list, json_ptr
9
#include "logging.hpp"               // for log_stream, error
10
#include "main.hpp"                  // for VERSION, NAME
11
#include "managed_io.hpp"            // for managed_io
12
#include "output.hpp"                // for DEV_NULL, output_files
13
#include "reports.hpp"               // for write_html_report
14
#include "reports_template_html.hpp" // for html_frequency_plot, html_demultiple...
15
#include "sequence_sets.hpp"         // for adapter_set
16
#include "simd.hpp"                  // for size_t
17
#include "statistics.hpp"            // for fastq_stats_ptr, fastq_statistics
18
#include "strutils.hpp"              // for format_percentage, format_rough...
19
#include "userconfig.hpp"            // for userconfig, ar_command, DEV_NULL
20
#include <algorithm>                 // for max
21
#include <cctype>                    // for toupper
22
#include <cerrno>                    // for errno
23
#include <cmath>                     // for fmod
24
#include <cstdint>                   // for uint64_t
25
#include <cstring>                   // for size_t, strerror
26
#include <iomanip>                   // for operator<<, setprecision, setw
27
#include <memory>                    // for __shared_ptr_access, shared_ptr
28
#include <sstream>                   // for ostringstream
29
#include <string>                    // for string, operator==, to_string
30
#include <string_view>               // for string_view
31
#include <utility>                   // for pair
32
#include <vector>                    // for vector
33

34
namespace adapterremoval {
35

36
namespace {
37

38
using fastq_stats_vec = std::vector<fastq_stats_ptr>;
39
using template_ptr = std::unique_ptr<html_template>;
40

41
//! Size chosen to allow fitting two pages side-by-side on a 1920 width display
42
const char* const FIGURE_WIDTH = "736";
43
//! Per figure width for two-column facet figures; approximate
44
const char* const FACET_WIDTH_2 = "351";
45
//! Per figure width for one-column facet figures; approximate
46
const char* const FACET_WIDTH_1 = FIGURE_WIDTH;
47

48
////////////////////////////////////////////////////////////////////////////////
49

50
/** Escapes a string that needs to be embedded in a JS */
51
std::string
52
json_encode(const std::string& s)
×
53
{
54
  return json_token::from_str(s)->to_string();
×
55
}
56

57
/** JSON escaped string */
58
std::string
59
operator""_json(const char* s, size_t length)
×
60
{
61
  return json_encode(std::string(s, length));
×
62
}
63

64
std::string
65
runtime_to_str(double seconds)
×
66
{
67
  std::ostringstream ss;
×
68

69
  if (seconds >= 3600.0) {
×
70
    ss << static_cast<size_t>(seconds / 3600.0) << " "
×
71
       << (seconds >= 7200.0 ? "hours, " : "hour, ") << std::setw(2);
×
72
  }
73

74
  if (seconds >= 60.0) {
×
75
    auto minutes = static_cast<size_t>(std::fmod(seconds, 3600.0) / 60.0);
×
76
    ss << minutes << " "
×
77
       << ((!minutes || minutes >= 120) ? "minutes" : "minute") << ", and "
×
78
       << std::setw(4);
×
79
  }
80

81
  ss << std::fixed << std::setprecision(1) << std::fmod(seconds, 60.0)
×
82
     << " seconds";
×
83

84
  return ss.str();
×
85
}
86

87
std::string
88
mean_of_bp_counts(const counts& count)
×
89
{
90
  auto reads = count.sum();
×
91
  auto bases = count.product();
×
92

93
  if (!reads) {
×
94
    return "NA";
×
95
  }
96

97
  if (bases % reads == 0) {
×
98
    return std::to_string(bases / reads) + " bp";
×
99
  }
100

101
  std::ostringstream ss;
×
102
  ss << std::fixed << std::setprecision(1)
×
103
     << (bases / static_cast<double>(reads)) << " bp";
×
104

105
  return ss.str();
×
106
}
107

108
/**
109
 * VEGA-lite will omit plots if there are no values; this function therefore
110
 * ensures that at least one value is written for a given measurement.
111
 */
112
template<typename T>
113
counts_tmpl<T>
114
require_values(counts_tmpl<T> r, T fallback = T())
×
115
{
116
  if (r.size()) {
×
117
    return r;
×
118
  }
119

120
  return counts_tmpl<T>({ fallback });
×
121
}
122

123
std::string
124
format_average_bases(const reads_and_bases& counts)
×
125
{
126
  const auto reads = counts.reads();
×
127

128
  if (reads) {
×
129
    return format_fraction(counts.bases(), reads, 1) + " bp";
×
130
  } else {
131
    return "NA";
×
132
  }
133
}
134

135
std::string
136
orientation_to_label(const sample_sequences& it)
×
137
{
138
  switch (it.orientation) {
×
139
    case barcode_orientation::unspecified:
×
140
      return {};
×
141
    case barcode_orientation::forward:
×
142
      return "+";
×
143
    case barcode_orientation::reverse:
×
144
      return "-";
×
145
    default:
×
146
      AR_FAIL("invalid barcode orientation");
×
147
  }
148
}
149

150
////////////////////////////////////////////////////////////////////////////////
151

152
class io_summary_writer
153
{
154
public:
155
  enum class io
156
  {
157
    input,
158
    output
159
  };
160

161
  io_summary_writer(std::ostream& output, const io type)
×
162
    : m_output(output)
×
163
    , m_type(type)
×
164

165
  {
166
  }
167

168
  void write_head(const std::string& title, const std::string& href)
×
169
  {
170
    html_summary_io_head().set_title(title).set_href(href).write(m_output);
×
171
  }
172

173
  void write_row(const std::string& title, const fastq_statistics& stats)
×
174
  {
175
    const auto n_reads = (m_type == io::input) ? stats.number_of_input_reads()
×
176
                                               : stats.number_of_output_reads();
×
177
    const auto total = stats.quality_dist().sum();
×
178

179
    html_summary_io_row()
×
180
      .set_name(title)
×
181
      .set_n_reads(format_rough_number(n_reads))
×
182
      .set_n_bases(format_rough_number(stats.length_dist().product()))
×
183
      .set_lengths(mean_of_bp_counts(stats.length_dist()))
×
184
      .set_q30(format_percentage(stats.quality_dist().sum(30), total))
×
185
      .set_q20(format_percentage(stats.quality_dist().sum(20), total))
×
186
      .set_ns(format_percentage(stats.nucleotides_pos('N').sum(), total))
×
187
      .set_gc(format_percentage(stats.nucleotides_gc_pos().sum(), total))
×
188
      .write(m_output);
×
189
  }
190

191
  void write_tail() { html_summary_io_tail().write(m_output); }
×
192

193
private:
194
  std::ostream& m_output;
195
  io m_type;
196
};
197

198
std::string
199
build_base_qualities(const fastq_stats_vec& reads, const string_vec& names)
×
200
{
201
  json_list qualities;
×
202

203
  for (size_t i = 0; i < reads.size(); ++i) {
×
204
    const auto& stats = *reads.at(i);
×
205

206
    auto total_quality = stats.qualities_pos();
×
207
    auto total_bases = stats.nucleotides_pos();
×
208

209
    for (const auto nucleotide : ACGT::values) {
×
210
      const auto nucleotides = stats.nucleotides_pos(nucleotide);
×
211
      const auto quality = stats.qualities_pos(nucleotide);
×
212

213
      auto dict = qualities.dict();
×
214
      dict->str("read", names.at(i));
×
215
      dict->i64("offset", 1);
×
216
      dict->str("group", std::string(1, ::toupper(nucleotide)));
×
217
      dict->f64_vec("y", quality / nucleotides);
×
218
    }
219

220
    auto dict = qualities.dict();
×
221
    dict->str("read", names.at(i));
×
222
    dict->i64("offset", 1);
×
223
    dict->str("group", "Mean");
×
224

225
    // Ensure that values get written, to prevent the plot being omitted
226
    dict->f64_vec("y", require_values(total_quality / total_bases));
×
227
  }
228

229
  return qualities.to_string();
×
230
}
231

232
std::string
233
build_quality_distribution(const fastq_stats_vec& reads,
×
234
                           const string_vec& names)
235
{
236
  json_list data;
×
237

238
  for (size_t i = 0; i < reads.size(); ++i) {
×
239
    const auto& stats = reads.at(i);
×
240
    auto count = stats->quality_dist().trim();
×
241
    // A max that should give a uniform look to most data
242
    count.resize_up_to(44);
×
243

244
    const auto m = data.dict();
×
245
    m->str("group", names.at(i));
×
246
    m->i64("offset", 0);
×
247
    m->i64_vec("y", count);
×
248
  }
249

250
  return data.to_string();
×
251
}
252

253
std::string
254
build_base_content(const fastq_stats_vec& reads, const string_vec& names)
×
255
{
256
  json_list content;
×
257

258
  for (size_t i = 0; i < reads.size(); ++i) {
×
259
    const auto& stats = *reads.at(i);
×
260

261
    auto total_bases = stats.nucleotides_pos();
×
262

263
    for (const auto nucleotide : ACGTN::values) {
×
264
      const auto bases = stats.nucleotides_pos(nucleotide);
×
265

266
      const auto dict = content.dict();
×
267
      dict->str("read", names.at(i));
×
268
      dict->i64("offset", 1);
×
269
      dict->str("group", std::string(1, nucleotide));
×
270

271
      // Ensure that values get written, to prevent the plot being omitted
272
      dict->f64_vec("y", require_values(bases / total_bases));
×
273
    }
274

275
    {
×
276
      const auto gc_content = stats.nucleotides_gc_pos();
×
277
      auto dict = content.dict();
×
278
      dict->str("read", names.at(i));
×
279
      dict->i64("offset", 1);
×
280
      dict->str("group", "GC");
×
281

282
      // Ensure that values get written, to prevent the plot being omitted
283
      dict->f64_vec("y", require_values(gc_content / total_bases));
×
284
    }
285
  }
286

287
  return content.to_string();
×
288
}
289

290
////////////////////////////////////////////////////////////////////////////////
291
// Main sections
292

293
void
294
write_html_sampling_note(const userconfig& config,
×
295
                         const std::string& label,
296
                         const fastq_statistics& stats,
297
                         std::ostream& output)
298
{
299
  if (config.report_sample_rate < 1.0) {
×
300
    html_sampling_note()
×
301
      .set_label(label)
×
302
      .set_reads(format_rough_number((stats.number_of_sampled_reads())))
×
303
      .set_pct(format_percentage(stats.number_of_sampled_reads(),
×
304
                                 stats.number_of_input_reads()))
×
305
      .write(output);
×
306
  }
307
}
308

309
void
310
write_html_summary_section(const userconfig& config,
×
311
                           const statistics& stats,
312
                           std::ostream& output)
313
{
314
  html_head().set_title(config.report_title).write(output);
×
315

316
  html_body_start().set_title(config.report_title).write(output);
×
317

318
  // Basic information about the executable / call
319
  {
×
320
    html_summary()
×
321
      .set_date_and_time(userconfig::start_time)
×
322
      .set_version(VERSION)
×
323
      .set_command(shell_escape_command(config.args))
×
324
      .set_runtime(runtime_to_str(config.runtime()))
×
325
      .write(output);
×
326
  }
327

328
  fastq_statistics output_1;
×
329
  fastq_statistics output_2;
×
330
  fastq_statistics merged;
×
331
  fastq_statistics singleton;
×
332
  fastq_statistics discarded;
×
333

334
  for (const auto& it : stats.trimming) {
×
335
    output_1 += *it->read_1;
×
336
    output_2 += *it->read_2;
×
337
    merged += *it->merged;
×
338
    singleton += *it->singleton;
×
339
    discarded += *it->discarded;
×
340
  }
341

342
  if (config.paired_ended_mode) {
×
343
    // Summary statistics for input files
344
    {
×
345
      fastq_statistics totals;
×
346
      totals += *stats.input_1;
×
347
      totals += *stats.input_2;
×
348

349
      io_summary_writer summary(output, io_summary_writer::io::input);
×
350
      summary.write_head("Input", "summary-input");
×
351
      if (config.paired_ended_mode) {
×
352
        summary.write_row("Summary", totals);
×
353
        summary.write_row("File 1", *stats.input_1);
×
354
        summary.write_row("File 2", *stats.input_2);
×
355
      }
356
      summary.write_tail();
×
357

358
      write_html_sampling_note(config, "input", totals, output);
×
359
    }
360

361
    // Summary statistics for output files
362
    if (config.run_type != ar_command::report_only) {
×
363
      fastq_statistics totals;
×
364
      totals += output_1;
×
365
      totals += output_2;
×
366
      totals += merged;
×
367
      totals += singleton;
×
368
      // discarded reads not counted in the output
369
      // totals += discarded;
370

371
      io_summary_writer summary{ output, io_summary_writer::io::output };
×
372
      summary.write_head("Output", "summary-output");
×
373
      summary.write_row("Passed*", totals);
×
374
      if (config.paired_ended_mode) {
×
375
        summary.write_row("File 1", output_1);
×
376
        summary.write_row("File 2", output_2);
×
377

378
        if (config.is_read_merging_enabled()) {
×
379
          summary.write_row("Merged", merged);
×
380
        }
381

382
        if (config.is_any_filtering_enabled()) {
×
383
          summary.write_row("Singleton", singleton);
×
384
        }
385
      }
386

387
      if (config.is_any_filtering_enabled()) {
×
388
        summary.write_row("Discarded*", discarded);
×
389
      }
390
      summary.write_tail();
×
391

392
      write_html_sampling_note(config, "output", totals, output);
×
393

394
      // Note regarding passed / discarded reads
395
      html_output_footnote()
×
396
        .set_symbol("*")
×
397
        .set_html("The <b>Passed</b> column includes all read types except "
×
398
                  "for <b>Discarded</b> reads.")
399
        .write(output);
×
400
    }
401
  } else if (config.run_type == ar_command::report_only) {
×
402
    io_summary_writer summary{ output, io_summary_writer::io::input };
×
403
    summary.write_head("Input summary", "summary-input");
×
404
    summary.write_row("Input", *stats.input_1);
×
405
    summary.write_tail();
×
406

407
    write_html_sampling_note(config, "input", *stats.input_1, output);
×
408
  }
409

410
  else {
411
    io_summary_writer summary{ output, io_summary_writer::io::input };
×
412
    summary.write_head("Input/Output summary", "summary-input-output");
×
413
    summary.write_row("Input", *stats.input_1);
×
414
    summary.write_row("Output", output_1);
×
415
    if (config.is_any_filtering_enabled()) {
×
416
      summary.write_row("Discarded*", discarded);
×
417
    }
418
    summary.write_tail();
×
419

420
    fastq_statistics totals;
×
421
    totals += *stats.input_1;
×
422
    totals += output_1;
×
423

424
    write_html_sampling_note(config, "input/output", totals, output);
×
425

426
    if (config.is_any_filtering_enabled()) {
×
427
      // Note regarding discarded reads in output
428
      html_output_footnote()
×
429
        .set_symbol("*")
×
430
        .set_html("<b>Discarded</b> reads are not included in the "
×
431
                  "<b>Output</b> column.")
432
        .write(output);
×
433
    }
434
  }
435
}
436

437
//! Trimming statistics
438
struct trimming_stats
439
{
440
  size_t id;
441
  //! Processing stage relative to adapter trimming (pre, X, post)
442
  std::string stage;
443
  //! Row label 1 (step)
444
  std::string label_1;
445
  //! Row label 1 (sub-step)
446
  std::string label_2;
447
  //! Whether or not this step is enabled by command-line options
448
  bool enabled;
449
  //! Number of reads/bases trimmed/filtered
450
  reads_and_bases count;
451
};
452

453
void
454
write_html_trimming_stats(std::ostream& output,
×
455
                          const std::vector<trimming_stats>& stats,
456
                          const reads_and_bases& totals)
457
{
458
  size_t n_processing_steps = 0;
×
459
  size_t n_processing_steps_on = 0;
×
460
  size_t n_filtering_steps = 0;
×
461
  size_t n_filtering_steps_on = 0;
×
462

463
  size_t last_id = -1;
×
464
  size_t last_enabled = -1;
×
465
  for (const auto& it : stats) {
×
466
    if (it.id != last_id) {
×
467
      if (it.stage == "Processing") {
×
468
        n_processing_steps++;
×
469
      } else if (it.stage == "Filtering") {
×
470
        n_filtering_steps++;
×
471
      }
472

473
      last_id = it.id;
×
474
    }
475

476
    if (it.enabled && it.id != last_enabled) {
×
477
      if (it.stage == "Processing") {
×
478
        n_processing_steps_on++;
×
479
      } else if (it.stage == "Filtering") {
×
480
        n_filtering_steps_on++;
×
481
      }
482

483
      last_enabled = it.id;
×
484
    }
485
  }
486

487
  html_summary_trimming_head().write(output);
×
488

489
  std::string previous_stage;
×
490
  std::string previous_label_1;
×
491

492
  for (const auto& it : stats) {
×
493
    if (it.enabled) {
×
494
      const auto label_1 = it.label_1 == previous_label_1 ? "" : it.label_1;
×
495
      const auto stage = it.stage == previous_stage ? "" : it.stage;
×
496

497
      previous_stage = it.stage;
×
498
      previous_label_1 = it.label_1;
×
499

500
      html_summary_trimming_row()
×
501
        .set_stage(stage)
×
502
        .set_label_1(label_1)
×
503
        .set_label_2(it.label_2)
×
504
        .set_reads(format_rough_number(it.count.reads()))
×
505
        .set_pct_reads(format_percentage(it.count.reads(), totals.reads()))
×
506
        .set_bases(format_rough_number(it.count.bases()))
×
507
        .set_pct_bases(format_percentage(it.count.bases(), totals.bases()))
×
508
        .set_avg_bases(format_average_bases(it.count))
×
509
        .write(output);
×
510
    }
511
  }
512

513
  html_summary_trimming_tail()
×
514
    .set_n_enabled_filt(std::to_string(n_filtering_steps_on))
×
515
    .set_n_total_filt(std::to_string(n_filtering_steps))
×
516
    .set_n_enabled_proc(std::to_string(n_processing_steps_on))
×
517
    .set_n_total_proc(std::to_string(n_processing_steps))
×
518
    .write(output);
×
519
}
520

521
//! Filtering statistics
522
struct filtering_stats
523
{
524
  //! Filtering step
525
  std::string label;
526
  //! Whether or not this step is enabled by command-line options
527
  bool enabled;
528
  //! Number of reads/bases trimmed/filtered
529
  reads_and_bases count;
530
};
531

532
reads_and_bases
533
summarize_input(const fastq_stats_ptr& ptr)
×
534
{
535
  const auto n_bases = ptr->length_dist().product();
×
536
  AR_REQUIRE(n_bases >= 0);
×
537

538
  return reads_and_bases{ ptr->number_of_input_reads(),
×
539
                          static_cast<uint64_t>(n_bases) };
540
}
541

542
void
543
build_polyx_trimming_rows(std::vector<trimming_stats>& out,
×
544
                          const std::string& polyx_nucleotides,
545
                          const indexed_count<ACGT>& reads,
546
                          const indexed_count<ACGT>& bases,
547
                          const size_t id)
548
{
549
  for (const auto nucleotide : ACGT::values) {
×
550
    out.push_back(
×
551
      { id,
552
        "Processing",
553
        "Poly-X tails",
554
        std::string(1, nucleotide),
555
        polyx_nucleotides.find(nucleotide) != std::string::npos,
×
556
        reads_and_bases(reads.get(nucleotide), bases.get(nucleotide)) });
×
557
  }
558

559
  out.push_back({ id,
×
560
                  "Processing",
561
                  "Poly-X tails",
562
                  "*",
563
                  polyx_nucleotides.size() > 1,
×
564
                  reads_and_bases(reads.sum(), bases.sum()) });
×
565
}
566

567
void
568
write_html_processing_section(const userconfig& config,
×
569
                              const statistics& stats,
570
                              std::ostream& output)
571
{
572
  trimming_statistics totals;
×
573
  for (const auto& it : stats.trimming) {
×
574
    totals += *it;
×
575
  }
576

577
  uint64_t adapter_reads = 0;
×
578
  uint64_t adapter_bases = 0;
×
579

580
  for (size_t i = 0; i < config.samples.adapters().size(); ++i) {
×
581
    adapter_reads += totals.adapter_trimmed_reads.get(i);
×
582
    adapter_bases += totals.adapter_trimmed_bases.get(i);
×
583
  }
584

585
  const auto total_input =
×
586
    summarize_input(stats.input_1) + summarize_input(stats.input_2);
×
587

588
  reads_and_bases total_output;
×
589
  for (const auto& it : stats.trimming) {
×
590
    total_output += summarize_input(it->read_1);
×
591
    total_output += summarize_input(it->read_2);
×
592
    total_output += summarize_input(it->singleton);
×
593
    total_output += summarize_input(it->merged);
×
594
  }
595

596
  // Trimming steps prior to adapter trimming
597
  size_t step_id = 0;
×
598
  std::vector<trimming_stats> trimming = {
×
599
    { step_id++, "Input", "Raw reads", "-", true, total_input },
×
600
    { step_id++,
×
601
      "Processing",
602
      "Terminal bases",
603
      "-",
604
      config.is_terminal_base_pre_trimming_enabled(),
×
605
      totals.terminal_pre_trimmed },
606
  };
607

608
  build_polyx_trimming_rows(trimming,
×
609
                            config.pre_trim_poly_x,
×
610
                            totals.poly_x_pre_trimmed_reads,
611
                            totals.poly_x_pre_trimmed_bases,
612
                            step_id++);
613

614
  trimming.push_back({ step_id++,
×
615
                       "Processing",
616
                       "Adapters",
617
                       "-",
618
                       config.is_adapter_trimming_enabled(),
×
619
                       reads_and_bases(adapter_reads, adapter_bases) });
620

621
  trimming.push_back({ step_id++,
×
622
                       "Processing",
623
                       "Merging",
624
                       "-",
625
                       config.is_read_merging_enabled(),
×
626
                       totals.reads_merged });
627

628
  trimming.push_back({ step_id++,
×
629
                       "Processing",
630
                       "Terminal bases",
631
                       "-",
632
                       config.is_terminal_base_post_trimming_enabled(),
×
633
                       totals.terminal_post_trimmed });
634

635
  build_polyx_trimming_rows(trimming,
×
636
                            config.post_trim_poly_x,
×
637
                            totals.poly_x_post_trimmed_reads,
638
                            totals.poly_x_post_trimmed_bases,
639
                            step_id++);
640

641
  trimming.push_back({ step_id++,
×
642
                       "Processing",
643
                       "Low quality bases",
644
                       "-",
645
                       config.is_low_quality_trimming_enabled(),
×
646
                       totals.low_quality_trimmed });
647

648
  trimming.push_back({ step_id++,
×
649
                       "Filtering",
650
                       "Short reads",
651
                       "-",
652
                       config.is_short_read_filtering_enabled(),
×
653
                       totals.filtered_min_length });
654

655
  trimming.push_back({ step_id++,
×
656
                       "Filtering",
657
                       "Long reads",
658
                       "-",
659
                       config.is_long_read_filtering_enabled(),
×
660
                       totals.filtered_max_length });
661
  trimming.push_back({ step_id++,
×
662
                       "Filtering",
663
                       "Ambiguous bases",
664
                       "-",
665
                       config.is_ambiguous_base_filtering_enabled(),
×
666
                       totals.filtered_ambiguous });
667
  trimming.push_back({ step_id++,
×
668
                       "Filtering",
669
                       "Mean quality",
670
                       "-",
671
                       config.is_mean_quality_filtering_enabled(),
×
672
                       totals.filtered_mean_quality });
673
  trimming.push_back({ step_id++,
×
674
                       "Filtering",
675
                       "Low complexity reads",
676
                       "-",
677
                       config.is_low_complexity_filtering_enabled(),
×
678
                       totals.filtered_low_complexity });
679

680
  trimming.push_back(
×
681
    { step_id++, "Output", "Filtered reads", "-", true, total_output });
×
682

683
  write_html_trimming_stats(output, trimming, total_input);
×
684
}
685

686
void
687
write_html_section_title(const std::string& title, std::ostream& output)
×
688
{
689
  html_h2_tag().set_title(title).set_href(to_lower(title)).write(output);
×
690
}
691

692
void
693
write_html_io_section(const userconfig& config,
×
694
                      std::ostream& output,
695
                      const std::string& title,
696
                      fastq_stats_vec statistics,
697
                      string_vec names,
698
                      const fastq_stats_ptr& merged = fastq_stats_ptr())
699
{
700
  AR_REQUIRE(statistics.size() == names.size());
×
701

702
  write_html_section_title(title, output);
×
703

704
  const char* dynamic_width =
×
705
    config.paired_ended_mode || merged ? FACET_WIDTH_2 : FACET_WIDTH_1;
×
706

707
  html_plot_title()
×
708
    .set_href(to_lower(title) + "-position-qualities")
×
709
    .set_title("Position quality distribution")
×
710
    .write(output);
×
711
  html_facet_line_plot()
×
712
    .set_x_axis(config.is_read_merging_enabled() && merged ? "null"
×
713
                                                           : "Position"_json)
714
    .set_y_axis("Phred score"_json)
×
715
    .set_width(dynamic_width)
×
716
    .set_values(build_base_qualities(statistics, names))
×
717
    .write(output);
×
718

719
  if (config.is_read_merging_enabled() && merged) {
×
720
    html_facet_line_plot()
×
721
      .set_x_axis("Position"_json)
×
722
      .set_y_axis("Phred score"_json)
×
723
      .set_width(FIGURE_WIDTH)
×
724
      .set_values(build_base_qualities({ merged }, { "Merged" }))
×
725
      .write(output);
×
726
  }
727

728
  html_plot_title()
×
729
    .set_href(to_lower(title) + "-nucleotide-content")
×
730
    .set_title("Nucleotide content")
×
731
    .write(output);
×
732
  html_facet_line_plot()
×
733
    .set_x_axis(config.is_read_merging_enabled() && merged ? "null"
×
734
                                                           : "Position"_json)
735
    .set_y_axis("Frequency"_json)
×
736
    .set_width(dynamic_width)
×
737
    .set_values(build_base_content(statistics, names))
×
738
    .write(output);
×
739

740
  if (config.is_read_merging_enabled() && merged) {
×
741
    html_facet_line_plot()
×
742
      .set_x_axis("Position"_json)
×
743
      .set_y_axis("Frequency"_json)
×
744
      .set_width(FIGURE_WIDTH)
×
745
      .set_values(build_base_content({ merged }, { "Merged" }))
×
746
      .write(output);
×
747

748
    // Subsequent plots should include merged reads
749
    names.push_back("Merged");
×
750
    statistics.push_back(merged);
×
751
  }
752

753
  html_plot_title()
×
754
    .set_href(to_lower(title) + "-quality-scores")
×
755
    .set_title("Quality score distribution")
×
756
    .write(output);
×
757
  html_frequency_plot()
×
758
    .set_x_axis("Phred score"_json)
×
759
    .set_y_axis("Frequency"_json)
×
760
    .set_width(FIGURE_WIDTH)
×
761
    .set_values(build_quality_distribution(statistics, names))
×
762
    .write(output);
×
763

764
  {
×
765
    json_list data;
×
766

767
    for (size_t i = 0; i < statistics.size(); ++i) {
×
768
      const auto m = data.dict();
×
769
      m->str("group", names.at(i));
×
770
      m->i64("offset", 0);
×
771
      m->f64_vec("y", statistics.at(i)->gc_content());
×
772
    }
773

774
    html_plot_title()
×
775
      .set_href(to_lower(title) + "-gc-content")
×
776
      .set_title("GC Content")
×
777
      .write(output);
×
778
    html_frequency_plot()
×
779
      .set_x_axis("%GC"_json)
×
780
      .set_y_axis("Frequency"_json)
×
781
      .set_width(FIGURE_WIDTH)
×
782
      .set_values(data.to_string())
×
783
      .write(output);
×
784
  }
785
}
786

787
void
788
write_html_input_section(const userconfig& config,
×
789
                         const statistics& stats,
790
                         std::ostream& output)
791
{
792
  fastq_stats_vec stats_vec = { stats.input_1 };
×
793
  string_vec names = { "File 1" };
×
794

795
  if (config.paired_ended_mode) {
×
796
    stats_vec.push_back(stats.input_2);
×
797
    names.emplace_back("File 2");
×
798
  }
799

800
  write_html_io_section(config,
×
801
                        output,
802
                        "Input",
803
                        std::move(stats_vec),
804
                        std::move(names));
805
}
806

807
void
808
write_html_analyses_section(const userconfig& config,
×
809
                            const statistics& stats,
810
                            std::ostream& output)
811

812
{
813
  write_html_section_title("Analyses", output);
×
814

815
  // Insert size distribution
816
  if (config.paired_ended_mode) {
×
817
    counts insert_sizes;
×
818
    for (const auto& it : stats.trimming) {
×
819
      insert_sizes += it->insert_sizes;
×
820
    }
821

822
    json_list samples;
×
823
    const auto sample = samples.dict();
×
824
    sample->str("group", "insert_sizes");
×
825
    sample->i64("offset", 0);
×
826
    sample->i64_vec("y", insert_sizes);
×
827

828
    // FIXME: Specify "identified reads" when in demultiplexing mode and
829
    // correct format_percentage to merged / n_identified.
830
    std::ostringstream ss;
×
831
    ss << "Insert sizes inferred for "
×
832
       << format_percentage(insert_sizes.sum(),
×
833
                            stats.input_1->number_of_input_reads())
×
834
       << " of reads";
×
835

836
    html_plot_title()
×
837
      .set_href("analyses-insert-sizes")
×
838
      .set_title("Insert-size distribution")
×
839
      .write(output);
×
840
    html_plot_sub_title().set_sub_title(ss.str()).write(output);
×
841
    html_frequency_plot()
×
842
      .set_x_axis("Insert size"_json)
×
843
      .set_y_axis("Frequency"_json)
×
844
      .set_legend("null")
×
845
      .set_width(FIGURE_WIDTH)
×
846
      .set_values(samples.to_string())
×
847
      .write(output);
×
848

849
    if (config.run_type == ar_command::report_only) {
×
850
      html_output_note()
×
851
        .set_text(
×
852
          "Insert size distribution inferred using adapter-free alignments.")
853
        .write(output);
×
854
    }
855
  }
856

857
  if (config.report_duplication) {
×
858
    AR_REQUIRE(stats.duplication_1 && stats.duplication_2);
×
859
    const auto dupes_1 = stats.duplication_1->summarize();
×
860
    const auto dupes_2 = stats.duplication_2->summarize();
×
861
    const auto mean_uniq_frac = (dupes_1.unique_frac + dupes_2.unique_frac) / 2;
×
862

863
    const auto to_percent = [](double value) {
×
864
      std::ostringstream os;
×
865
      os << std::fixed << std::setprecision(1) << (value * 100.0) << " %";
×
866
      return os.str();
×
867
    };
868

869
    html_duplication_head().write(output);
×
870
    if (config.paired_ended_mode) {
×
871
      html_duplication_body_pe()
×
872
        .set_pct_unique(to_percent(mean_uniq_frac))
×
873
        .set_pct_unique_1(to_percent(dupes_1.unique_frac))
×
874
        .set_pct_unique_2(to_percent(dupes_2.unique_frac))
×
875
        .write(output);
×
876
    } else {
877
      html_duplication_body_se()
×
878
        .set_pct_unique(to_percent(dupes_1.unique_frac))
×
879
        .write(output);
×
880
    }
881

882
    const auto add_line = [](json_list& list,
×
883
                             std::string_view read,
884
                             std::string_view group,
885
                             const std::vector<std::string>& labels,
886
                             const rates& values) {
887
      AR_REQUIRE(labels.size() == values.size());
×
888
      for (size_t i = 0; i < labels.size(); ++i) {
×
889
        auto dict = list.dict();
×
890
        dict->str("read", read);
×
891
        dict->str("group", group);
×
892
        dict->str("x", labels.at(i));
×
893
        dict->f64("y", values.get(i));
×
894
      }
895
    };
896

897
    json_list data;
×
898
    const auto add_lines = [add_line, &data](const decltype(dupes_1)& s,
×
899
                                             std::string_view label) {
900
      add_line(data, label, "All", s.labels, s.total_sequences);
×
901
      add_line(data, label, "Unique", s.labels, s.unique_sequences);
×
902
    };
903

904
    add_lines(dupes_1, "File 1");
×
905
    if (config.paired_ended_mode) {
×
906
      add_lines(dupes_2, "File 2");
×
907
    }
908

909
    html_duplication_plot()
×
910
      .set_width(config.paired_ended_mode ? FACET_WIDTH_2 : FACET_WIDTH_1)
×
911
      .set_values(data.to_string())
×
912
      .write(output);
×
913
  }
914

915
  // Consensus adapter sequence inference
916
  if (config.paired_ended_mode && config.run_type == ar_command::report_only) {
×
917
    AR_REQUIRE(stats.adapter_id);
×
918

919
    const auto adapter_1 = stats.adapter_id->adapter1.summarize();
×
920
    const auto adapter_2 = stats.adapter_id->adapter2.summarize();
×
921

922
    // Consensus adapter sequences
923
    {
×
924
      const auto reference_adapters =
×
925
        config.samples.adapters().to_read_orientation().front();
×
926
      std::string reference_adapter_1{ reference_adapters.first };
×
927
      std::string reference_adapter_2{ reference_adapters.second };
×
928

929
      html_consensus_adapter_head()
×
930
        .set_overlapping_pairs(
×
931
          format_rough_number(stats.adapter_id->aligned_pairs))
×
932
        .set_pairs_with_adapters(
×
933
          format_rough_number(stats.adapter_id->pairs_with_adapters))
×
934
        .write(output);
×
935

936
      html_consensus_adapter_table()
×
937
        .set_name_1("--adapter1")
×
938
        .set_reference_1(reference_adapter_1)
×
939
        .set_alignment_1(adapter_1.compare_with(reference_adapter_1))
×
940
        .set_consensus_1(adapter_1.adapter().sequence())
×
941
        .set_qualities_1(adapter_1.adapter().qualities())
×
942
        .set_name_2("--adapter2")
×
943
        .set_reference_2(reference_adapter_2)
×
944
        .set_alignment_2(adapter_2.compare_with(reference_adapter_2))
×
945
        .set_consensus_2(adapter_2.adapter().sequence())
×
946
        .set_qualities_2(adapter_2.adapter().qualities())
×
947
        .write(output);
×
948
    }
949

950
    // Top N most common 5' kmers in adapter fragments
951
    {
×
952
      const auto& top_kmers_1 = adapter_1.top_kmers();
×
953
      const auto& top_kmers_2 = adapter_2.top_kmers();
×
954

955
      html_consensus_adapter_kmer_head()
×
956
        .set_n_kmers(std::to_string(consensus_adapter_stats::top_n_kmers))
×
957
        .set_kmer_length(std::to_string(consensus_adapter_stats::kmer_length))
×
958
        .write(output);
×
959

960
      const auto kmers = std::max(top_kmers_1.size(), top_kmers_2.size());
×
961
      for (size_t i = 0; i < kmers; ++i) {
×
962
        html_consensus_adapter_kmer_row row;
×
963
        row.set_index(std::to_string(i + 1));
×
964

965
        if (top_kmers_1.size() > i) {
×
966
          const auto& kmer = top_kmers_1.at(i);
×
967

968
          row.set_kmer_1(kmer.first)
×
969
            .set_count_1(format_rough_number(kmer.second))
×
970
            .set_pct_1(format_percentage(kmer.second, adapter_1.total_kmers()));
×
971
        }
972

973
        if (top_kmers_2.size() > i) {
×
974
          const auto& kmer = top_kmers_2.at(i);
×
975

976
          row.set_kmer_2(kmer.first)
×
977
            .set_count_2(format_rough_number(kmer.second))
×
978
            .set_pct_2(format_percentage(kmer.second, adapter_2.total_kmers()));
×
979
        }
980

981
        row.write(output);
×
982
      }
983

984
      html_consensus_adapter_kmer_tail().write(output);
×
985
    }
986
  }
987
}
988

989
void
990
write_html_demultiplexing_barplot(const userconfig& config,
×
991
                                  const statistics& stats,
992
                                  std::ostream& output)
993
{
994
  json_list data;
×
995

996
  const size_t input_reads = stats.input_1->number_of_input_reads() +
×
997
                             stats.input_2->number_of_input_reads();
×
998

999
  for (size_t i = 0; i < config.samples.size(); ++i) {
×
1000
    const auto& sample = config.samples.at(i);
×
1001

1002
    for (size_t j = 0; j < sample.size(); ++j) {
×
1003
      auto count = stats.demultiplexing->samples.at(i).get(j);
×
1004

1005
      const auto& sequences = sample.at(j);
×
1006
      std::string key{ sequences.barcode_1 };
×
1007
      if (!sequences.barcode_2.empty()) {
×
1008
        key.push_back('-');
×
1009
        key.append(sequences.barcode_2);
×
1010
      }
1011

1012
      auto m = data.dict();
×
1013
      m->i64("n", j + 1);
×
1014
      m->str("barcodes", key);
×
1015

1016
      if (sequences.orientation != barcode_orientation::unspecified) {
×
1017
        m->str("orientation", orientation_to_label(sequences));
×
1018
      }
1019

1020
      m->str("sample", sample.name());
×
1021

1022
      if (input_reads) {
×
1023
        m->f64("pct", (100.0 * count) / input_reads);
×
1024
      } else {
1025
        m->null("pct");
×
1026
      }
1027
    }
1028
  }
1029

1030
  html_plot_title()
×
1031
    .set_href("demux-samples")
×
1032
    .set_title("Samples identified")
×
1033
    .write(output);
×
1034
  html_bar_plot()
×
1035
    .set_x_axis("Samples"_json)
×
1036
    .set_y_axis("Percent"_json)
×
1037
    .set_width(FIGURE_WIDTH)
×
1038
    .set_values(data.to_string())
×
1039
    .write(output);
×
1040
}
1041

1042
void
1043
write_html_demultiplexing_table(const userconfig& config,
×
1044
                                const statistics& stats,
1045
                                std::ostream& output,
1046
                                const bool multiple_barcodes,
1047
                                const bool mixed_orientation)
1048
{
1049
  const size_t input_reads = stats.input_1->number_of_input_reads() +
×
1050
                             stats.input_2->number_of_input_reads();
×
1051

1052
  html_demultiplexing_table_head()
×
1053
    .set_orientation(mixed_orientation ? "<th></th>" : "")
×
1054
    .write(output);
×
1055

1056
  {
×
1057
    const size_t unidentified = stats.demultiplexing->unidentified;
×
1058

1059
    fastq_statistics total;
×
1060
    total += *stats.demultiplexing->unidentified_stats_1;
×
1061
    total += *stats.demultiplexing->unidentified_stats_2;
×
1062

1063
    const auto output_reads = total.length_dist().sum();
×
1064
    const auto output_bp = total.nucleotides_pos().sum();
×
1065

1066
    html_demultiplexing_row()
×
1067
      .set_name("<b>Unidentified</b>")
×
1068
      .set_sample_pct(format_percentage(unidentified, input_reads, 2))
×
1069
      .set_reads(format_rough_number(output_reads))
×
1070
      .set_bp(format_rough_number(output_bp))
×
1071
      .set_length(mean_of_bp_counts(total.length_dist()))
×
1072
      .set_gc(format_percentage(total.nucleotides_gc_pos().sum(), output_bp))
×
1073
      .set_orientation(mixed_orientation ? "<td></td>" : "")
×
1074
      .write(output);
×
1075
  }
1076

1077
  size_t sample_idx = 0;
×
1078
  for (const auto& sample : config.samples) {
×
1079
    const auto& output_stats = *stats.trimming.at(sample_idx);
×
1080
    const auto& barcode_counts = stats.demultiplexing->samples.at(sample_idx);
×
1081
    const auto sample_reads = barcode_counts.sum();
×
1082

1083
    fastq_statistics total;
×
1084

1085
    total += *output_stats.read_1;
×
1086
    total += *output_stats.read_2;
×
1087
    total += *output_stats.merged;
×
1088
    total += *output_stats.singleton;
×
1089
    // Not included in overview:
1090
    // total += *sample.discarded;
1091

1092
    const auto output_reads = total.length_dist().sum();
×
1093
    const auto output_bp = total.nucleotides_pos().sum();
×
1094

1095
    html_demultiplexing_row row;
×
1096
    if (sample.size() < 2) {
×
1097
      const auto& it = sample.at(0);
×
1098
      row.set_barcode_1(std::string{ it.barcode_1 })
×
1099
        .set_barcode_2(std::string{ it.barcode_2 });
×
1100

1101
      if (mixed_orientation) {
×
1102
        row.set_orientation("<td>" + orientation_to_label(it) + "</td>");
×
1103
      }
1104
    } else {
1105
      const auto cell = "<i>" + std::to_string(sample.size()) + " barcodes</i>";
×
1106
      row.set_barcode_1(cell).set_barcode_2(cell);
×
1107

1108
      if (mixed_orientation) {
×
1109
        row.set_orientation("<td></td>");
×
1110
      }
1111
    }
1112

1113
    row.set_n(std::to_string(sample_idx + 1))
×
1114
      .set_name(sample.name())
×
1115
      .set_sample_pct(format_percentage(sample_reads, input_reads, 2))
×
1116
      .set_reads(format_rough_number(output_reads))
×
1117
      .set_bp(format_rough_number(output_bp))
×
1118
      .set_length(mean_of_bp_counts(total.length_dist()))
×
1119
      .set_gc(format_percentage(total.nucleotides_gc_pos().sum(), output_bp))
×
1120
      .write(output);
×
1121

1122
    if (sample.size() > 1) {
×
1123
      const auto total = barcode_counts.sum();
×
1124

1125
      for (size_t j = 0; j < sample.size(); j++) {
×
1126
        const auto& it = sample.at(j);
×
1127
        const auto count = barcode_counts.get(j);
×
1128

1129
        html_demultiplexing_barcode_row row;
×
1130
        row.set_barcode_1(std::string{ it.barcode_1 })
×
1131
          .set_barcode_2(std::string{ it.barcode_2 })
×
1132
          .set_barcode_pct_row(format_percentage(count, total, 2));
×
1133

1134
        if (mixed_orientation) {
×
1135
          row.set_orientation("<td>" + orientation_to_label(it) + "</td>");
×
1136
        }
1137

1138
        row.write(output);
×
1139
      }
1140
    }
1141

1142
    ++sample_idx;
×
1143
  }
1144

1145
  html_demultiplexing_table_tail().write(output);
×
1146

1147
  if (multiple_barcodes || mixed_orientation) {
×
1148
    html_demultiplexing_toggle().write(output);
×
1149
  }
1150
}
1151

1152
void
1153
write_html_demultiplexing_section(const userconfig& config,
×
1154
                                  const statistics& stats,
1155
                                  std::ostream& output)
1156

1157
{
1158
  bool multiple_barcodes = false;
×
1159
  bool mixed_orientation = false;
×
1160
  for (const auto& sample : config.samples) {
×
1161
    multiple_barcodes |= sample.size() > 1;
×
1162
    for (const auto& it : sample) {
×
1163
      mixed_orientation |= it.orientation != barcode_orientation::unspecified;
×
1164
    }
1165
  }
1166

1167
  write_html_section_title("Demultiplexing", output);
×
1168
  html_demultiplexing_head().write(output);
×
1169
  write_html_demultiplexing_barplot(config, stats, output);
×
1170
  write_html_demultiplexing_table(config,
×
1171
                                  stats,
1172
                                  output,
1173
                                  multiple_barcodes,
1174
                                  mixed_orientation);
1175
}
1176

1177
void
1178
write_html_output_section(const userconfig& config,
×
1179
                          const statistics& stats,
1180
                          std::ostream& output)
1181

1182
{
1183
  fastq_stats_vec stats_vec;
×
1184
  string_vec names;
×
1185

1186
  auto merged = std::make_shared<fastq_statistics>();
×
1187

1188
  {
×
1189
    auto output_1 = std::make_shared<fastq_statistics>();
×
1190
    auto output_2 = std::make_shared<fastq_statistics>();
×
1191
    auto singleton = std::make_shared<fastq_statistics>();
×
1192
    auto discarded = std::make_shared<fastq_statistics>();
×
1193

1194
    for (const auto& it : stats.trimming) {
×
1195
      *output_1 += *it->read_1;
×
1196
      *output_2 += *it->read_2;
×
1197
      *merged += *it->merged;
×
1198
      *singleton += *it->singleton;
×
1199
      *discarded += *it->discarded;
×
1200
    }
1201

1202
    stats_vec.push_back(output_1);
×
1203
    names.emplace_back("Output 1");
×
1204

1205
    if (config.paired_ended_mode) {
×
1206
      stats_vec.push_back(output_2);
×
1207
      names.emplace_back("Output 2");
×
1208

1209
      if (config.is_any_filtering_enabled()) {
×
1210
        stats_vec.push_back(singleton);
×
1211
        names.emplace_back("Singleton");
×
1212
      }
1213
    }
1214

1215
    if (config.is_any_filtering_enabled()) {
×
1216
      stats_vec.push_back(discarded);
×
1217
      names.emplace_back("Discarded");
×
1218
    }
1219
  }
1220

1221
  write_html_io_section(config,
×
1222
                        output,
1223
                        "Output",
1224
                        std::move(stats_vec),
1225
                        std::move(names),
1226
                        merged);
1227
}
1228

1229
} // namespace
1230

1231
////////////////////////////////////////////////////////////////////////////////
1232

1233
bool
1234
write_html_report(const userconfig& config,
×
1235
                  const statistics& stats,
1236
                  const std::string& filename)
1237
{
1238
  if (filename == DEV_NULL) {
×
1239
    // User disabled the report
1240
    return true;
1241
  }
1242

1243
  std::ostringstream output;
×
1244

1245
  write_html_summary_section(config, stats, output);
×
1246

1247
  if (config.run_type != ar_command::demultiplex_only &&
×
1248
      config.run_type != ar_command::report_only) {
1249
    write_html_processing_section(config, stats, output);
×
1250
  }
1251

1252
  write_html_input_section(config, stats, output);
×
1253

1254
  if (config.paired_ended_mode || config.report_duplication ||
×
1255
      config.run_type == ar_command::report_only) {
×
1256
    write_html_analyses_section(config, stats, output);
×
1257
  }
1258

1259
  if (config.is_demultiplexing_enabled()) {
×
1260
    write_html_demultiplexing_section(config, stats, output);
×
1261
  }
1262

1263
  if (config.run_type != ar_command::report_only) {
×
1264
    write_html_output_section(config, stats, output);
×
1265
  }
1266

1267
  html_body_end().write(output);
×
1268

1269
  try {
×
1270
    managed_writer writer{ filename };
×
1271
    writer.write(output.str());
×
1272
    writer.close();
×
NEW
1273
  } catch (const io_error& error) {
×
1274
    log::error() << "Error writing JSON report to '" << filename << "':\n"
×
1275
                 << indent_lines(error.what());
×
1276
    return false;
×
1277
  }
×
1278

1279
  return true;
×
1280
}
1281

1282
} // namespace adapterremoval
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