File: alignment_test.cpp

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/*************************************************************************\
 * AdapterRemoval - cleaning next-generation sequencing reads            *
 *                                                                       *
 * Copyright (C) 2011 by Stinus Lindgreen - stinus@binf.ku.dk            *
 * Copyright (C) 2014 by Mikkel Schubert - mikkelsch@gmail.com           *
 *                                                                       *
 * If you use the program, please cite the paper:                        *
 * S. Lindgreen (2012): AdapterRemoval: Easy Cleaning of Next Generation *
 * Sequencing Reads, BMC Research Notes, 5:337                           *
 * http://www.biomedcentral.com/1756-0500/5/337/                         *
 *                                                                       *
 * This program is free software: you can redistribute it and/or modify  *
 * it under the terms of the GNU General Public License as published by  *
 * the Free Software Foundation, either version 3 of the License, or     *
 * (at your option) any later version.                                   *
 *                                                                       *
 * This program is distributed in the hope that it will be useful,       *
 * but WITHOUT ANY WARRANTY; without even the implied warranty of        *
 * MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE.  See the         *
 * GNU General Public License for more details.                          *
 *                                                                       *
 * You should have received a copy of the GNU General Public License     *
 * along with this program.  If not, see <http://www.gnu.org/licenses/>. *
\*************************************************************************/
#include <limits>
#include <sstream>
#include <vector>

#include "testing.hpp"
#include "alignment.hpp"
#include "fastq.hpp"


#define TEST_ALIGNMENT_SETTER(TYPE, NAME) \
    ALN& NAME(TYPE value) \
    { \
        info.adapter_id = 0; \
        info.NAME = value; \
        return *this; \
    }


namespace ar
{


bool operator==(const alignment_info& first, const alignment_info& second)
{
    return (first.offset == second.offset)
        && (first.score == second.score)
        && (first.length == second.length)
        && (first.n_mismatches == second.n_mismatches)
        && (first.n_ambiguous == second.n_ambiguous)
        && (first.adapter_id == second.adapter_id);
}


struct ALN
{
    ALN() : info() {}

    TEST_ALIGNMENT_SETTER(int, score);
    TEST_ALIGNMENT_SETTER(int, offset);
    TEST_ALIGNMENT_SETTER(size_t, length);
    TEST_ALIGNMENT_SETTER(size_t, n_mismatches);
    TEST_ALIGNMENT_SETTER(size_t, n_ambiguous);
    TEST_ALIGNMENT_SETTER(int, adapter_id);

    operator alignment_info() {
        return info;
    }

    alignment_info info;
};


bool operator==(const alignment_info& first, const ALN& second)
{
    return first == second.info;
}


std::ostream& operator<<(std::ostream& stream, const alignment_info& aln)
{
    std::vector<std::string> labels = {
        "score", "offset", "length",
        "n_mismatches", "n_ambiguous",
        "adapter_id"
    };

    std::vector<int64_t> values = {
        static_cast<int64_t>(aln.score),
        static_cast<int64_t>(aln.offset),
        static_cast<int64_t>(aln.length),
        static_cast<int64_t>(aln.n_mismatches),
        static_cast<int64_t>(aln.n_ambiguous),
        static_cast<int64_t>(aln.adapter_id),
    };

    stream << "alignment_info(";

    bool any_streamed = false;
    for (size_t i = 0; i < labels.size(); ++i) {
        if (values.at(i)) {
            if (any_streamed) {
                stream << "\n               ";
            }

            stream << labels.at(i) << " = " << values.at(i);
            any_streamed = true;
        }
    }

    return stream << ")";
}


void REQUIRE_TRUNCATED_PE_IS_UNCHANGED(const alignment_info& alignment,
                                      const fastq& record1,
                                      const fastq& record2)
{
    fastq tmp_record1 = record1;
    fastq tmp_record2 = record2;
    REQUIRE(truncate_paired_ended_sequences(alignment, tmp_record1, tmp_record2) == 0);
    REQUIRE(tmp_record1 == record1);
    REQUIRE(tmp_record2 == record2);
}


fastq_pair_vec create_adapter_vec(const fastq& pcr1, const fastq& pcr2 = fastq())
{
    fastq_pair_vec adapters;
    adapters.push_back(fastq_pair(pcr1, pcr2));
    return adapters;
}


std::random_device g_seed;
std::mt19937 g_rng_instance(g_seed());
std::mt19937* g_rng(&g_rng_instance);


///////////////////////////////////////////////////////////////////////////////
// Cases for SE alignments (a = read 1, b = adapter, o = overlap):
//  1. No overlap = aaaaaa bbbbbb
//  2. Partial overlap = aaaaaooobbbbbb
//  3. Complete overlap = ooooooooo
//  4. a contains b = aaaaoooooo
//  5. b contains a = ooooobbbbb
//  6. a extends past b = aaaaoooooaaaa
//  7. b extends past a = bbbbooooobbbb
//  8. both extends past other = bbbbooooaaaa


///////////////////////////////////////////////////////////////////////////////
// Case 1: No overlap between sequences:
//         AAAAAAAAAAA
//                       BBBBBBBBBBBBBBB
// Expected result = suboptimal alignment or no overlap

TEST_CASE("SE: Unalignable sequence yields default alignment", "[alignment::single_end]")
{
    const fastq record("Rec",  "AAAA", "!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Rec", "TTTT", "!!!!"));

    REQUIRE(align_single_ended_sequence(record, adapters, 0) == ALN());
}


TEST_CASE("SE: Random sequences yields suboptimal alignment", "[alignment::single_end]")
{
    const fastq record("Rec",  "ACGTAGTA",  "!!!!!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Rec", "TGAGACGGT", "!!!!!!!!!"));

    REQUIRE(align_single_ended_sequence(record, adapters, 0) == ALN().offset(6)
                                                                     .length(2)
                                                                     .n_mismatches(1));
}


///////////////////////////////////////////////////////////////////////////////
// Case 2: Partial overlap between sequences:
//         AAAAAAAAAAA
//                BBBBBBBBBBBBBBB
// Expected result = optimal alignment between ends
TEST_CASE("SE: Partial alignment between ends", "[alignment::single_end]")
{
    const fastq record("Rec",  "ACGTAGTAA", "123457890");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Rec", "AGTAAGGT",  "!!!!!!!!"));
    const alignment_info expected = ALN().score(5).offset(4).length(5);
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);

    fastq tmp_record = record;
    truncate_single_ended_sequence(result, tmp_record);
    REQUIRE(tmp_record == fastq("Rec", "ACGT", "1234"));
}


TEST_CASE("SE: Partial alignment with mismatches between ends", "[alignment::single_end]")
{
    const fastq record("Rec",  "ACGTAGTAA", "123457890");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Rec", "AGGAAGGT",  "!!!!!!!!"));
    const alignment_info expected = ALN().score(3).offset(4).length(5).n_mismatches(1);
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);

    fastq tmp_record = record;
    truncate_single_ended_sequence(result, tmp_record);
    REQUIRE(tmp_record == fastq("Rec", "ACGT", "1234"));
}


TEST_CASE("SE: Partial alignment with ambiguous between ends", "[alignment::single_end]")
{
    const fastq record("Rec",  "ACGTAGTAA", "123457890");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Rec", "AGNAAGGT",  "!!!!!!!!"));
    const alignment_info expected = ALN().score(4).offset(4).length(5).n_ambiguous(1);
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);

    fastq tmp_record = record;
    truncate_single_ended_sequence(result, tmp_record);
    REQUIRE(tmp_record == fastq("Rec", "ACGT", "1234"));
}


///////////////////////////////////////////////////////////////////////////////
// Case 3: Complete overlap between sequences:
//         AAAAAAAAAAA
//         BBBBBBBBBBB
// Expected result = Optimal alignment involving all bases

TEST_CASE("SE: Completely overlapping sequences", "[alignment::single_end]")
{
    const fastq record("Rec", "ACGTAGTA", "!!!!!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(record);
    const alignment_info expected = ALN().score(8).length(8);
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);

    fastq tmp_record = record;
    truncate_single_ended_sequence(result, tmp_record);
    REQUIRE(tmp_record == fastq("Rec", "", ""));
}


TEST_CASE("SE: Completely overlapping sequences with mismatches", "[alignment::single_end]")
{
    const fastq record("Rec", "ACGTAGTA", "!!!!!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Rec", "GCGTAGTA", "!!!!!!!!"));
    const alignment_info expected = ALN().score(6).length(8).n_mismatches(1);
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);

    fastq tmp_record = record;
    truncate_single_ended_sequence(result, tmp_record);
    REQUIRE(tmp_record == fastq("Rec", "", ""));
}


TEST_CASE("SE: Completely overlapping sequences with mismatches and ambiguous", "[alignment::single_end]")
{
    const fastq record("Rec", "ACGTAGTA", "!!!!!!!!");
    const fastq_pair_vec adapter = create_adapter_vec(fastq("Rec", "GCGTAGTN", "!!!!!!!!"));
    const alignment_info expected = ALN().score(5).length(8).n_mismatches(1).n_ambiguous(1);
    const alignment_info result = align_single_ended_sequence(record, adapter, 0);
    REQUIRE(result == expected);

    fastq tmp_record = record;
    truncate_single_ended_sequence(result, tmp_record);
    REQUIRE(tmp_record == fastq("Rec", "", ""));
}


///////////////////////////////////////////////////////////////////////////////
// Cases 4 and 5: Sequence A completely contains sequence B (and vice versa)
//         AAAAAAAAAAA  AAAA
//               BBBBB  BBBBBBBBBB
// Expected result = Optimal alignment involving all bases of the shortest read

TEST_CASE("Complete adapter inside sequence", "[alignment::single_end]")
{
    const fastq record("Rec", "ACGTAGTA", "ABCDEFGH");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Adp",   "TAGTA", "!!!!!"));
    const alignment_info expected = ALN().score(5).offset(3).length(5);
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);

    fastq tmp_record = record;
    truncate_single_ended_sequence(result, tmp_record);
    REQUIRE(tmp_record == fastq("Rec", "ACG", "ABC"));
}


TEST_CASE("Complete adapter inside sequence with mismatch", "[alignment::single_end]")
{
    const fastq record("Rec", "ACGTAGTA", "ABCDEFGH");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Adp", "TATTA", "!!!!!"));
    const alignment_info expected = ALN().score(3).offset(3).length(5).n_mismatches(1);
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);

    fastq tmp_record = record;
    truncate_single_ended_sequence(result, tmp_record);
    REQUIRE(tmp_record == fastq("Rec", "ACG", "ABC"));
}


TEST_CASE("Complete adapter inside sequence with ambiguous", "[alignment::single_end]")
{
    const fastq record("Rec", "ACGTAGTA", "ABCDEFGH");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Adp", "TAGNA", "!!!!!"));
    const alignment_info expected = ALN().score(4).offset(3).length(5).n_ambiguous(1);
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);

    fastq tmp_record = record;
    truncate_single_ended_sequence(result, tmp_record);
    REQUIRE(tmp_record == fastq("Rec", "ACG", "ABC"));
}


TEST_CASE("Complete sequence inside adapter", "[alignment::single_end]")
{
    const fastq record("Rec",  "ACGT", "!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Adp", "ACGTAGTA", "!!!!!!!!"));
    const alignment_info expected = ALN().score(4).length(4);
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);

    fastq tmp_record = record;
    truncate_single_ended_sequence(result, tmp_record);
    REQUIRE(tmp_record == fastq("Rec", "", ""));
}


TEST_CASE("Complete sequence inside adapter with mismatches", "[alignment::single_end]")
{
    const fastq record("Rec",  "ACGT",     "!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Adp", "GCGTAGTA", "!!!!!!!!"));
    const alignment_info expected = ALN().score(2).length(4).n_mismatches(1);
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);

    fastq tmp_record = record;
    truncate_single_ended_sequence(result, tmp_record);
    REQUIRE(tmp_record == fastq("Rec", "", ""));
}


TEST_CASE("Complete sequence inside adapter with ambiguous", "[alignment::single_end]")
{
    const fastq record("Rec",  "ACGT", "!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Adp", "ACGNAGTA", "!!!!!!!!"));
    const alignment_info expected = ALN().score(3).length(4).n_ambiguous(1);
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);

    fastq tmp_record = record;
    truncate_single_ended_sequence(result, tmp_record);
    REQUIRE(tmp_record == fastq("Rec", "", ""));
}


///////////////////////////////////////////////////////////////////////////////
// Cases 6 and 7: Sequence A extends past sequence B (and vice versa)
//         AAAAAAAAAAAAAA    AAAA
//               BBBBB     BBBBBBBBBBBBB

TEST_CASE("Sequence extends past adapter", "[alignment::single_end]")
{
    const fastq record("Rec", "ACGTAGTATA", "0123456789");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Adp", "AGTA", "!!!!"));
    const alignment_info expected = ALN().score(4).offset(4).length(4);
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);

    fastq tmp_record = record;
    truncate_single_ended_sequence(result, tmp_record);
    REQUIRE(tmp_record == fastq("Rec", "ACGT", "0123"));
}


TEST_CASE("Sequence extends past adapter, no shift", "[alignment::single_end]")
{
    const fastq record("Rec", "CGTA", "#!%%");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Adp", "ACGTAGTATA", "!!!!!!!!!!"));
    const alignment_info expected = ALN().score(1).offset(3).length(1);
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);

    fastq tmp_record = record;
    truncate_single_ended_sequence(result, tmp_record);
    REQUIRE(tmp_record == fastq("Rec", "CGT", "#!%"));
}


TEST_CASE("Sequence extends past adapter, shift 1", "[alignment::single_end]")
{
    const fastq record("Rec", "CGTA", "#!%%");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Adp", "ACGTAGTATA", "!!!!!!!!!!"));
    const alignment_info expected = ALN().score(4).offset(-1).length(4);
    const alignment_info result = align_single_ended_sequence(record, adapters, 1);
    REQUIRE(result == expected);

    fastq tmp_record = record;
    truncate_single_ended_sequence(result, tmp_record);
    REQUIRE(tmp_record == fastq("Rec", "", ""));
}


///////////////////////////////////////////////////////////////////////////////
// Cases 8: Both sequences extends past the other
//                      AAAAAAAAAAAAAAAAAA
//               BBBBBBBBBBBBBBBBBB
// Expected result = Optimal alignment involving all overlapping bases

TEST_CASE("Sequence and adapter extends past each other", "[alignment::single_end]")
{
    const fastq record("Rec", "ACGTAGTATATAGT", "!!!!!!!!!!!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Adp", "CCGAACGTAGTATA", "!!!!!!!!!!!!!!"));
    const alignment_info expected = ALN().score(10).offset(-4).length(10);
    const alignment_info result = align_single_ended_sequence(record, adapters, 4);
    REQUIRE(result == expected);

    fastq tmp_record = record;
    truncate_single_ended_sequence(result, tmp_record);
    REQUIRE(tmp_record == fastq("Rec", "", ""));
}


///////////////////////////////////////////////////////////////////////////////
// Empty sequence or adapter

TEST_CASE("Empty sequence alignment", "[alignment::single_end]")
{
    const fastq record;
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Adp", "CCGAACGTAGTATA", "!!!!!!!!!!!!!!"));
    const alignment_info expected;
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);
}


TEST_CASE("Empty adapter alignment", "[alignment::single_end]")
{
    const fastq record("Rec", "ACGTAGTATATAGT", "!!!!!!!!!!!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq());
    const alignment_info expected;
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);
}

///////////////////////////////////////////////////////////////////////////////
// Misc

TEST_CASE("Lower than possible shift is allowed", "[alignment::single_end]")
{
    const fastq record("Rec",  "AAAA", "!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("Adp", "TTTT", "!!!!"));
    const alignment_info expected;
    const alignment_info result = align_single_ended_sequence(record, adapters, -10);
    REQUIRE(result == expected);
}


TEST_CASE("Only adapter 1 is used", "[alignment::single_end]")
{
    const fastq_pair_vec adapters = create_adapter_vec(fastq("barcode", "AAA", "JJJ"),
                                                       fastq("barcode", "TTTAAA", "JJJJJJ"));
    const fastq record("Rec",  "CCCCTTTAAA", "0987654321");
    const alignment_info expected = ALN().score(3).offset(7).length(3);
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);
}


TEST_CASE("Best matching adapter is returned: First", "[alignment::single_end]")
{
    fastq_pair_vec adapters;
    adapters.push_back(fastq_pair(fastq("adapter", "TGCTGC", "JJJJJJ"), fastq()));
    adapters.push_back(fastq_pair(fastq("adapter", "TGCTGA", "JJJJJJ"), fastq()));

    const fastq record("Read", "TAGTCGCTATGCTGC", "!!!!!!!!!103459");
    const alignment_info expected = ALN().score(6).offset(9).length(6);
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);
}


TEST_CASE("Best matching adapter returned: Second", "[alignment::single_end]")
{
    fastq_pair_vec adapters;
    adapters.push_back(fastq_pair(fastq("adapter", "TGCTGA", "JJJJJJ"), fastq()));
    adapters.push_back(fastq_pair(fastq("adapter", "TGCTGC", "JJJJJJ"), fastq()));

    const fastq record("Read", "TAGTCGCTATGCTGC", "!!!!!!!!!103459");
    const alignment_info expected = ALN().score(6).offset(9).length(6).adapter_id(1);
    const alignment_info result = align_single_ended_sequence(record, adapters, 0);
    REQUIRE(result == expected);
}


TEST_CASE("Best matching adapter returned: Neither", "[alignment::single_end]")
{
    fastq_pair_vec barcodes;
    barcodes.push_back(fastq_pair(fastq("barcode", "AAAAAA", "JJJJJJ"), fastq()));
    barcodes.push_back(fastq_pair(fastq("barcode", "CCCCCC", "JJJJJJ"), fastq()));

    const fastq record = fastq("Read", "AACTGTACGTAGTT", "!!!!!!10345923");
    const alignment_info result = align_single_ended_sequence(record, barcodes, 0);
    REQUIRE(result == ALN());
}



///////////////////////////////////////////////////////////////////////////////
// Case 1 PE: No overlap between sequences:
//         AAAAAAAAAAA
//                       BBBBBBBBBBBBBBB
// Expected result = suboptimal alignment or no overlap

TEST_CASE("Unalignable sequence pair", "[alignment::paired_end]")
{
    const fastq record1("Rec",  "AAAA", "!!!!");
    const fastq record2("Rec", "TTTT", "!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("PCR1", "CGCTGA", "!!!!!!"),
                                                       fastq("PCR2", "TGTAC",  "!!!!!"));
    const alignment_info expected;
    const alignment_info result = align_paired_ended_sequences(record1, record2, adapters, 0);
    REQUIRE(result == expected);
    REQUIRE_TRUNCATED_PE_IS_UNCHANGED(result, record1, record2);
}


TEST_CASE("No overlap in sequence pair", "[alignment::paired_end]")
{
    const fastq record1("Rec",  "ACGTAGTA",  "!!!!!!!!");
    const fastq record2("Rec", "TGAGACGGT", "!!!!!!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("PCR1", "CGCTGA", "!!!!!!"),
                                                       fastq("PCR2", "TGTAC",  "!!!!!"));
    const alignment_info expected = ALN().offset(6).length(2).n_mismatches(1);
    const alignment_info result = align_paired_ended_sequences(record1, record2, adapters, 0);
    REQUIRE(result == expected);
    REQUIRE_TRUNCATED_PE_IS_UNCHANGED(result, record1, record2);
}


///////////////////////////////////////////////////////////////////////////////
// Case 2 PE: Partial overlap between sequences:
//         AAAAAAAAAAA
//                BBBBBBBBBBBBBBB
// Expected result = optimal alignment between ends

TEST_CASE("Partial overlap in sequence pair", "[alignment::paired_end]")
{
    const fastq record1("Rec", "ACGTAGTAA", "!!!!!!!!!");
    const fastq record2("Rec", "AGTAAGGT",  "!!!!!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("PCR1", "CGCTGA", "!!!!!!"),
                                                       fastq("PCR2", "TGTAC",  "!!!!!"));
    const alignment_info expected = ALN().score(5).offset(4).length(5);
    const alignment_info result = align_paired_ended_sequences(record1, record2, adapters, 0);
    REQUIRE(result == expected);
    REQUIRE_TRUNCATED_PE_IS_UNCHANGED(result, record1, record2);
}


///////////////////////////////////////////////////////////////////////////////
// Case 3 PE: Complete overlap between sequences:
//         AAAAAAAAAAA
//         BBBBBBBBBBB
// Expected result = Optimal alignment involving all bases

TEST_CASE("Completely overlapping sequence pair", "[alignment::paired_end]")
{
    const fastq record1("Rec", "ACGTAGTA", "!!!!!!!!");
    const fastq record2 = record1;
    const fastq_pair_vec adapters = create_adapter_vec(fastq("PCR1", "CGCTGA", "!!!!!!"),
                                                       fastq("PCR2", "TGTAC",  "!!!!!"));
    const alignment_info expected = ALN().score(8).length(8);
    const alignment_info result = align_paired_ended_sequences(record1, record2, adapters, 0);
    REQUIRE(result == expected);
    REQUIRE_TRUNCATED_PE_IS_UNCHANGED(result, record1, record2);
}


///////////////////////////////////////////////////////////////////////////////
// Cases 4 and 5 PE: Sequence A completely contains sequence B (and vice versa)
//         AAAAAAAAAAA  AAAA
//               BBBBB  BBBBBBBBBB
// Expected result = Optimal alignment involving all bases of the shortest read

TEST_CASE("Sequence A contains sequence B", "[alignment::paired_end]")
{
    const fastq record1("Rec1", "ACGTAGTA", "!!!!!!!!");
    const fastq record2("Rec2", "TAGTA", "!!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("PCR1", "CGCTGA", "!!!!!!"),
                                                       fastq("PCR2", "TGTAC",  "!!!!!"));
    const alignment_info expected = ALN().score(5).offset(3).length(5);
    const alignment_info result = align_paired_ended_sequences(record1, record2, adapters, 0);
    REQUIRE(result == expected);
    REQUIRE_TRUNCATED_PE_IS_UNCHANGED(result, record1, record2);
}


TEST_CASE("Sequence B contains sequence A", "[alignment::paired_end]")
{
    const fastq record1("Rec1", "ACGT", "!!!!");
    const fastq record2("Rec2", "ACGTAGTA", "!!!!!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("PCR1", "CGCTGA", "!!!!!!"),
                                                       fastq("PCR2", "TGTAC",  "!!!!!"));
    const alignment_info expected = ALN().score(4).length(4);
    const alignment_info result = align_paired_ended_sequences(record1, record2, adapters, 0);
    REQUIRE(result == expected);
    REQUIRE_TRUNCATED_PE_IS_UNCHANGED(result, record1, record2);
}


///////////////////////////////////////////////////////////////////////////////
// Cases 6 and 7 PE: Sequence A extends past sequence B (and vice versa)
//         AAAAAAAAAAAAAA    AAAA
//               BBBBB     BBBBBBBBBBBBB
TEST_CASE("Sequence A extends past sequence B", "[alignment::paired_end]")
{
    const fastq record1("Rec1",  "ACGTAGTACG", "!!!!!!!!!!");
    const fastq record2("Rec2",      "AGTA",   "!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("PCR1", "CGCTGA", "!!!!!!"),
                                                       fastq("PCR2", "TGTAC",  "!!!!!"));
    const alignment_info expected = ALN().score(6).offset(4).length(6);
    const alignment_info result = align_paired_ended_sequences(record1, record2, adapters, 0);
    REQUIRE(result == expected);

    fastq tmp_record1 = record1;
    fastq tmp_record2 = record2;
    REQUIRE(truncate_paired_ended_sequences(result, tmp_record1, tmp_record2) == 1);
    REQUIRE(tmp_record1 == fastq("Rec1", "ACGTAGTA", "!!!!!!!!"));
    REQUIRE(tmp_record2 == fastq("Rec2", "AGTA", "!!!!"));
}


TEST_CASE("Sequence B extends past sequence A", "[alignment::paired_end]")
{
    const fastq record1("Rec1",   "CGTA",      "!!!!");
    const fastq record2("Rec2", "ACCGTAGTAT", "!!!!!!!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("PCR1", "CGCTGA", "!!!!!!"),
                                                       fastq("PCR2", "TGTAC",  "!!!!!"));
    const alignment_info expected = ALN().score(6).offset(-2).length(6);
    const alignment_info result = align_paired_ended_sequences(record1, record2, adapters, 0);
    REQUIRE(result == expected);

    fastq tmp_record1 = record1;
    fastq tmp_record2 = record2;
    REQUIRE(truncate_paired_ended_sequences(result, tmp_record1, tmp_record2) == 1);
    REQUIRE(tmp_record1 == fastq("Rec1", "CGTA", "!!!!"));
    REQUIRE(tmp_record2 == fastq("Rec2", "CGTAGTAT", "!!!!!!!!"));
}

///////////////////////////////////////////////////////////////////////////////
// Cases 8 PE: Both sequences extends past the other
//                      AAAAAAAAAAAAAAAAAA
//               BBBBBBBBBBBBBBBBBB
// Expected result = Optimal alignment involving all overlapping bases

TEST_CASE("Sequences extends past each other", "[alignment::paired_end]")
{
    const fastq record1("Rec1",     "ACGTAGTATACGCT", "!!!!!!!!!!!!!!");
    const fastq record2("Rec2", "GTACACGTAGTATA",     "!!!!!!!!!!!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("PCR1", "CGCTGA", "!!!!!!"),
                                                       fastq("PCR2", "TGTAC",  "!!!!!"));
    const alignment_info expected = ALN().score(18).offset(-4).length(18);
    const alignment_info result = align_paired_ended_sequences(record1, record2, adapters, 0);
    REQUIRE(result == expected);

    fastq tmp_record1 = record1;
    fastq tmp_record2 = record2;
    REQUIRE(truncate_paired_ended_sequences(result, tmp_record1, tmp_record2) == 2);
    REQUIRE(tmp_record1 == fastq("Rec1", "ACGTAGTATA", "!!!!!!!!!!"));
    REQUIRE(tmp_record2 == fastq("Rec2", "ACGTAGTATA", "!!!!!!!!!!"));
}


TEST_CASE("Adapter only sequences", "[alignment::paired_end]")
{
    const fastq record1("Rec1", "CCCGAC", "!!!!!!");
    const fastq record2("Rec2", "ATGCCTT", "!!!!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("PCR1", "CCCGACCCGT", "!!!!!!!!!!"),
                                                       fastq("PCR2", "AAGATGCCTT", "!!!!!!!!!!"));
    const alignment_info expected = ALN().score(13).offset(-7).length(13);
    const alignment_info result = align_paired_ended_sequences(record1, record2, adapters, 0);

    REQUIRE(result == expected);
}


TEST_CASE("Adapter only sequences, with missing base", "[alignment::paired_end]")
{
    // Test the case where both reads are adapters, but are missing a single base
    // Normally, alignments that do not invovle read1 vs read2 are skipped, but
    // missing bases may cause some alignments to be missed.
    const fastq record1("Rec1", "CCGACC", "!!!!!!");
    const fastq record2("Rec2", "ATGCCT", "!!!!!!");
    const fastq_pair_vec adapters = create_adapter_vec(fastq("PCR1", "CCCGACCCGT", "!!!!!!!!!!"),
                                                       fastq("PCR2", "AAGATGCCTT", "!!!!!!!!!!"));

    // Sub-optimal alignment:
    //   aagatgccttCCGACC
    //          ATGCCTcccgacccgt
    const alignment_info expected_1 = ALN().score(1).offset(-3).length(9).n_mismatches(4);
    const alignment_info result_1 = align_paired_ended_sequences(record1, record2, adapters, 0);
    REQUIRE(result_1 == expected_1);
    // Optimal alignment, only possible with shift
    //   aagatgccttCCGACC
    //      ATGCCTcccgacccgt
    const alignment_info expected_2 = ALN().score(11).offset(-7).length(13).n_mismatches(1);
    const alignment_info result_2 = align_paired_ended_sequences(record1, record2, adapters, 1);
    REQUIRE(result_2 == expected_2);
}


///////////////////////////////////////////////////////////////////////////////
//

TEST_CASE("Invalid alignment", "[alignment::paired_end]")
{
    fastq record1("Rec", "", "");
    fastq record2("Rec", "", "");
    const alignment_info alignment = ALN().offset(1);

    REQUIRE_THROWS_AS(truncate_paired_ended_sequences(alignment, record1, record2), std::invalid_argument);
}


///////////////////////////////////////////////////////////////////////////////
// Merging of reads using the original method implemented in AdapterRemoval

TEST_CASE("Collapse partial overlap", "[alignment::collapse]")
{
    sequence_merger merger(g_rng);
    fastq record1("Rec1", "ATATTATA", "01234567");
    fastq record2("Rec2", "NNNNACGT", "ABCDEFGH");
    const alignment_info alignment = ALN().offset(4);
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 0);
    const fastq collapsed_expected = fastq("Rec1", "ATATTATAACGT", "01234567EFGH");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Collapse complete overlap", "[alignment::collapse]")
{
    sequence_merger merger(g_rng);
    fastq record1("Rec1",  "ATATTATAA", "JJJJJJJJJ");
    fastq record2("Rec2", "AATATTATA", "JJJJJJJJJ");
    const alignment_info alignment = ALN().offset(-1);
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 2);
    const fastq collapsed_expected = fastq("Rec1", "ATATTATA", "wwwwwwww", FASTQ_ENCODING_SAM);
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Collapse complete overlap for mate 1", "[alignment::collapse]")
{
    sequence_merger merger(g_rng);
    fastq record1("Rec1", "ATATTATAG", "JJJJJJJJJ");
    fastq record2("Rec2", "ATATTATA",  "JJJJJJJJ");
    const alignment_info alignment = ALN();
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 1);
    const fastq collapsed_expected = fastq("Rec1", "ATATTATA", "wwwwwwww", FASTQ_ENCODING_SAM);
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Collapse complete overlap for mate 2", "[alignment::collapse]")
{
    sequence_merger merger(g_rng);
    fastq record1("Rec1",  "ATATTATA", "JJJJJJJJ");
    fastq record2("Rec2", "AATATTATA", "JJJJJJJJJ");
    const alignment_info alignment = ALN().offset(-1);
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 1);
    const fastq collapsed_expected = fastq("Rec1", "ATATTATA", "wwwwwwww", FASTQ_ENCODING_SAM);
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Unequal sequence lengths, mate 1 shorter", "[alignment::collapse]")
{
    sequence_merger merger(g_rng);
    fastq record1("Rec1", "ATA", "012");
    fastq record2("Rec2", "NNNNACGT", "ABCDEFGH");
    const alignment_info alignment = ALN().offset(3);
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 0);
    const fastq collapsed_expected = fastq("Rec1", "ATANNNNACGT", "012ABCDEFGH");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Unequal sequence lengths, mate 1 shorter, mate 2 extends past", "[alignment::collapse]")
{
    sequence_merger merger(g_rng);
    fastq record1("Rec1", "ATA", "012");
    fastq record2("Rec2", "AANNNNACGT", "90ABCDEFGH");
    const alignment_info alignment = ALN().offset(-2);
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 1);
    const fastq collapsed_expected = fastq("Rec1", "ATANACGT", "012DEFGH");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Unequal sequence lengths, mate 2 shorter", "[alignment::collapse]")
{
    sequence_merger merger(g_rng);
    fastq record1("Rec1", "ATATTATA", "01234567");
    fastq record2("Rec2", "ACG", "EFG");
    const alignment_info alignment = ALN().offset(8);
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 0);
    const fastq collapsed_expected = fastq("Rec1", "ATATTATAACG", "01234567EFG");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Ambiguous sites are filled from mate", "[alignment::collapse]")
{
    sequence_merger merger(g_rng);
    fastq record1("Rec1", "NNNNNNTATA", "0123456789");
    fastq record2("Rec2", "ACGTNNNNNN", "ABCDEFGHIJ");
    const alignment_info alignment;
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 0);
    const fastq collapsed_expected = fastq("Rec1", "ACGTNNTATA", "ABCD!!6789");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Identical nucleotides gets higher qualities", "[alignment::collapse]")
{
    sequence_merger merger(g_rng);
    fastq record1("Rec1", "GCATGATATA", "012345!0:A");
    fastq record2("Rec2", "TATATACAAC", "(3&?EFGHIJ");
    const alignment_info alignment = ALN().offset(6);
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 0);
    const fastq collapsed_expected = fastq("Rec1", "GCATGATATATACAAC", "012345(FBcEFGHIJ", FASTQ_ENCODING_SAM);
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Identical nucleotides gets higher qualities, no more than 41", "[alignment::collapse]")
{
    sequence_merger merger(g_rng);
    fastq record1("Rec1", "GCATGATATA", "0123456789");
    fastq record2("Rec2", "TATATACAAC", "ABCDEFGHIJ");
    const alignment_info alignment = ALN().offset(6);
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 0);
    const fastq collapsed_expected = fastq("Rec1", "GCATGATATATACAAC", "012345Z\\^`EFGHIJ", FASTQ_ENCODING_SAM);
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Higher quality nucleotide is selected", "[alignment::collapse]")
{
    sequence_merger merger(g_rng);
    fastq record1("Rec1", "GCATGAGCAT", "012345!0:A");
    fastq record2("Rec2", "TATATACAAC", "(3&?EFGHIJ");
    const alignment_info alignment = ALN().offset(6);
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 0);
    const fastq collapsed_expected = fastq("Rec1", "GCATGATAATTACAAC", "012345(%5%EFGHIJ");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Randomly select between different nucleotides with same quality #1", "[alignment::collapse]")
{
    const fastq record1("Rec1", "G", "1");
    const fastq record2("Rec2", "T", "1");
    const alignment_info alignment;
    std::seed_seq seed{1};
    std::mt19937 rng(seed);
    sequence_merger merger(&rng);

    const fastq collapsed_expected = fastq("Rec1", "G", "#");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Randomly select between different nucleotides with same quality #2", "[alignment::collapse]")
{
    const fastq record1("Rec1", "G", "1");
    const fastq record2("Rec2", "T", "1");
    const alignment_info alignment;
    std::seed_seq seed{2};
    std::mt19937 rng(seed);
    sequence_merger merger(&rng);

    const fastq collapsed_expected = fastq("Rec1", "T", "#");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Set conflicts to N/! if no RNG is provided", "[alignment::collapse]")
{
    const fastq record1("Rec1", "G", "1");
    const fastq record2("Rec2", "T", "1");
    const alignment_info alignment;
    sequence_merger merger;

    const fastq collapsed_expected = fastq("Rec1", "N", "!");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Offsets past the end throws", "[alignment::collapse]")
{
    sequence_merger merger(g_rng);
    const fastq record1("Rec1", "G", "1");
    const fastq record2("Rec2", "T", "1");
    const alignment_info alignment = ALN().offset(2);
    REQUIRE_THROWS_AS(merger.merge(alignment, record1, record2), std::invalid_argument);
}


TEST_CASE("Mate numbering is removed", "[alignment::collapse]")
{
    sequence_merger merger(g_rng);
    const fastq record1("Read/1", "ATATTATA", "01234567");
    const fastq record2("Read/2", "NNNNACGT", "ABCDEFGH");
    const alignment_info alignment = ALN().offset(4);
    const fastq collapsed_expected = fastq("Read", "ATATTATAACGT", "01234567EFGH");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);

    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Mate numbering removed, meta from 1 kept", "[alignment::collapse]")
{
    sequence_merger merger(g_rng);
    const fastq record1("Read/1 Meta1", "ATATTATA", "01234567");
    const fastq record2("Read/2 Meta2", "NNNNACGT", "ABCDEFGH");
    const alignment_info alignment = ALN().offset(4);
    const fastq collapsed_expected = fastq("Read Meta1", "ATATTATAACGT", "01234567EFGH");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);

    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Mate numbering removed, non-standard separator", "[alignment::collapse]")
{
    sequence_merger merger(g_rng);
    merger.set_mate_separator(':');

    const fastq record1("Read:1", "ATATTATA", "01234567");
    const fastq record2("Read:2", "NNNNACGT", "ABCDEFGH");
    const alignment_info alignment = ALN().offset(4);
    const fastq collapsed_expected = fastq("Read", "ATATTATAACGT", "01234567EFGH");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);

    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Mate numbering removed, non-standard separator, not set", "[alignment::collapse]")
{
    sequence_merger merger(g_rng);
    const fastq record1("Read:1", "ATATTATA", "01234567");
    const fastq record2("Read:2", "NNNNACGT", "ABCDEFGH");
    const alignment_info alignment = ALN().offset(4);
    const fastq collapsed_expected = fastq("Read:1", "ATATTATAACGT", "01234567EFGH");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);

    REQUIRE(collapsed_result == collapsed_expected);
}


///////////////////////////////////////////////////////////////////////////////
// Merging of reads using the newer, more conservative method

TEST_CASE("Conservative merge partial overlap", "[alignment::merge::conservative]")
{
    sequence_merger merger;
    merger.set_conservative(true);

    fastq record1("Rec1", "ATATTATA", "01234567");
    fastq record2("Rec2", "NNNNACGT", "ABCDEFGH");
    const alignment_info alignment = ALN().offset(4);
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 0);
    const fastq collapsed_expected = fastq("Rec1", "ATATTATAACGT", "01234567EFGH");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Conservative merge complete overlap", "[alignment::merge::conservative]")
{
    sequence_merger merger;
    merger.set_conservative(true);

    fastq record1("Rec1",  "ATATTATAA", "JJJJJJJJJ");
    fastq record2("Rec2", "AATATTATA", "JJJJJJJJJ");
    const alignment_info alignment = ALN().offset(-1);
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 2);
    const fastq collapsed_expected = fastq("Rec1", "ATATTATA", "JJJJJJJJ", FASTQ_ENCODING_SAM);
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Conservative merge complete overlap for mate 1", "[alignment::merge::conservative]")
{
    sequence_merger merger;
    merger.set_conservative(true);

    fastq record1("Rec1", "ATATTATAG", "JJJJJJJJJ");
    fastq record2("Rec2", "ATATTATA",  "JJJJJJJJ");
    const alignment_info alignment = ALN();
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 1);
    const fastq collapsed_expected = fastq("Rec1", "ATATTATA", "JJJJJJJJ", FASTQ_ENCODING_SAM);
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Conservative merge complete overlap for mate 2", "[alignment::merge::conservative]")
{
    sequence_merger merger;
    merger.set_conservative(true);

    fastq record1("Rec1",  "ATATTATA", "JJJJJJJJ");
    fastq record2("Rec2", "AATATTATA", "JJJJJJJJJ");
    const alignment_info alignment = ALN().offset(-1);
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 1);
    const fastq collapsed_expected = fastq("Rec1", "ATATTATA", "JJJJJJJJ", FASTQ_ENCODING_SAM);
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Conservative merge of unequal sequence lengths, mate 1 shorter", "[alignment::merge::conservative]")
{
    sequence_merger merger;
    merger.set_conservative(true);

    fastq record1("Rec1", "ATA", "012");
    fastq record2("Rec2", "NNNNACGT", "ABCDEFGH");
    const alignment_info alignment = ALN().offset(3);
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 0);
    const fastq collapsed_expected = fastq("Rec1", "ATANNNNACGT", "012ABCDEFGH");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Conservative merge of unequal sequence lengths, mate 1 shorter, mate 2 extends past", "[alignment::merge::conservative]")
{
    sequence_merger merger;
    merger.set_conservative(true);

    fastq record1("Rec1", "ATA", "012");
    fastq record2("Rec2", "AANNNNACGT", "90ABCDEFGH");
    const alignment_info alignment = ALN().offset(-2);
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 1);
    const fastq collapsed_expected = fastq("Rec1", "ATANACGT", "012DEFGH");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Conservative merge of unequal sequence lengths, mate 2 shorter", "[alignment::merge::conservative]")
{
    sequence_merger merger;
    merger.set_conservative(true);

    fastq record1("Rec1", "ATATTATA", "01234567");
    fastq record2("Rec2", "ACG", "EFG");
    const alignment_info alignment = ALN().offset(8);
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 0);
    const fastq collapsed_expected = fastq("Rec1", "ATATTATAACG", "01234567EFG");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Conservative merge of ambiguous sites filled from mate", "[alignment::merge::conservative]")
{
    sequence_merger merger;
    merger.set_conservative(true);

    fastq record1("Rec1", "NNNNNNTATA", "0123456789");
    fastq record2("Rec2", "ACGTNNNNNN", "ABCDEFGHIJ");
    const alignment_info alignment;
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 0);
    const fastq collapsed_expected = fastq("Rec1", "ACGTNNTATA", "ABCD!!6789");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Conservative merge of identical nucleotides, gets highest quality", "[alignment::merge::conservative]")
{
    sequence_merger merger;
    merger.set_conservative(true);

    fastq record1("Rec1", "GCATGATATA", "012345!0:A");
    fastq record2("Rec2", "TATATACAAC", "(3&?EFGHIJ");
    const alignment_info alignment = ALN().offset(6);
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 0);
    const fastq collapsed_expected = fastq("Rec1", "GCATGATATATACAAC", "012345(3:AEFGHIJ", FASTQ_ENCODING_SAM);
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}


TEST_CASE("Conservative merge of selects higher quality nucleotide", "[alignment::merge::conservative]")
{
    sequence_merger merger;
    merger.set_conservative(true);

    fastq record1("Rec1", "GCATGAGCAT", "012345!0:A");
    fastq record2("Rec2", "TATATACAAC", "(3&?EFGHIJ");
    const alignment_info alignment = ALN().offset(6);
    REQUIRE(truncate_paired_ended_sequences(alignment, record1, record2) == 0);
    const fastq collapsed_expected = fastq("Rec1", "GCATGATAATTACAAC", "012345($5#EFGHIJ");
    const fastq collapsed_result = merger.merge(alignment, record1, record2);
    REQUIRE(collapsed_result == collapsed_expected);
}

///////////////////////////////////////////////////////////////////////////////
// Adapter extraction

TEST_CASE("Extracting empty sequences yields empty sequences #1", "[alignment::extract_adapter_sequences]")
{
    const fastq expected_1 = fastq("read1", "", "");
    const fastq expected_2 = fastq("read2", "", "");
    fastq read1 = expected_1;
    fastq read2 = expected_2;
    extract_adapter_sequences(ALN(), read1, read2);
    REQUIRE(read1 == expected_1);
    REQUIRE(read2 == expected_2);
}


TEST_CASE("Extracting empty sequences yields empty sequences #2", "[alignment::extract_adapter_sequences]")
{
    const fastq expected_1 = fastq("read1", "", "");
    const fastq expected_2 = fastq("read2", "GGGGCC", "!!!!!!");
    fastq read1 = expected_1;
    fastq read2 = fastq("read2", "GGGGCC", "!!!!!!");
    extract_adapter_sequences(ALN(), read1, read2);
    REQUIRE(read1 == fastq("read1", "", ""));
    REQUIRE(read2 == fastq("read2", "", ""));
}


TEST_CASE("Extracting empty sequences yields empty sequences #3", "[alignment::extract_adapter_sequences]")
{
    const fastq expected_1 = fastq("read1", "AATTTT", "!!!!!!");
    const fastq expected_2 = fastq("read2", "", "");
    fastq read1 = fastq("read1", "", "");
    fastq read2 = expected_2;
    extract_adapter_sequences(ALN(), read1, read2);
    REQUIRE(read1 == fastq("read1", "", ""));
    REQUIRE(read2 == fastq("read2", "", ""));
}


TEST_CASE("Extracting empty sequences yields empty sequences #4", "[alignment::extract_adapter_sequences]")
{
    const fastq expected_1 = fastq("read1", "", "");
    const fastq expected_2 = fastq("read2", "", "");
    fastq read1 = expected_1;
    fastq read2 = expected_2;
    extract_adapter_sequences(ALN(), read1, read2);
    REQUIRE(read1 == fastq("read1", "", ""));
    REQUIRE(read2 == fastq("read2", "", ""));
}


TEST_CASE("Extracting with no alignment yields empty sequences", "[alignment::extract_adapter_sequences]")
{
    const fastq expected_1 = fastq("read1", "AATTTT", "!!!!!!");
    const fastq expected_2 = fastq("read2", "GGGGCC", "!!!!!!");
    fastq read1 = expected_1;
    fastq read2 = expected_2;
    extract_adapter_sequences(ALN(), read1, read2);
    REQUIRE(read1 == fastq("read1", "", ""));
    REQUIRE(read2 == fastq("read2", "", ""));
}


TEST_CASE("Extracting with partial overlap yields empty sequences", "[alignment::extract_adapter_sequences]")
{
    const fastq expected_1 = fastq("read1", "AATTTT", "!!!!!!");
    const fastq expected_2 = fastq("read2", "GGGGCC", "!!!!!!");
    fastq read1 = expected_1;
    fastq read2 = expected_2;
    extract_adapter_sequences(ALN().offset(2), read1, read2);
    REQUIRE(read1 == fastq("read1", "", ""));
    REQUIRE(read2 == fastq("read2", "", ""));
}


TEST_CASE("Extracting with complete overlap yields empty sequences", "[alignment::extract_adapter_sequences]")
{
    const fastq expected_1 = fastq("read1", "AATTTT", "!!!!!!");
    const fastq expected_2 = fastq("read2", "GGGGCC", "!!!!!!");
    fastq read1 = expected_1;
    fastq read2 = expected_2;
    extract_adapter_sequences(ALN(), read1, read2);
    REQUIRE(read1 == fastq("read1", "", ""));
    REQUIRE(read2 == fastq("read2", "", ""));
}


TEST_CASE("Extracting sequence 2 inside sequence 1 yields empty sequences", "[alignment::extract_adapter_sequences]")
{
    const fastq expected_1 = fastq("read1", "AATTTT", "!!!!!!");
    const fastq expected_2 = fastq("read2", "GGCC", "!!!!");
    fastq read1 = expected_1;
    fastq read2 = expected_2;
    extract_adapter_sequences(ALN().offset(2), read1, read2);
    REQUIRE(read1 == fastq("read1", "", ""));
    REQUIRE(read2 == fastq("read2", "", ""));
}


TEST_CASE("Extracting sequence 1 inside sequence 2 yields empty sequences", "[alignment::extract_adapter_sequences]")
{
    const fastq expected_1 = fastq("read1", "AATT", "!!!!");
    const fastq expected_2 = fastq("read2", "GGGGCC", "!!!!!!");
    fastq read1 = expected_1;
    fastq read2 = expected_2;
    extract_adapter_sequences(ALN(), read1, read2);
    REQUIRE(read1 == fastq("read1", "", ""));
    REQUIRE(read2 == fastq("read2", "", ""));
}


TEST_CASE("Extracting sequence 1 extending past sequence 2", "[alignment::extract_adapter_sequences]")
{
    const fastq expected_1 = fastq("read1", "AATTTTCC", "12345678");
    const fastq expected_2 = fastq("read2", "GGGGGG", "!!!!!!");
    fastq read1 = expected_1;
    fastq read2 = expected_2;
    extract_adapter_sequences(ALN(), read1, read2);
    REQUIRE(read1 == fastq("read1", "CC", "78"));
    REQUIRE(read2 == fastq("read2", "", ""));
}


TEST_CASE("Extracting sequence 2 extending past sequence 1", "[alignment::extract_adapter_sequences]")
{
    const fastq expected_1 = fastq("read1", "TTTTTT", "!!!!!!");
    const fastq expected_2 = fastq("read2", "AAGGGGGG", "12345678");
    fastq read1 = expected_1;
    fastq read2 = expected_2;
    extract_adapter_sequences(ALN().offset(-2), read1, read2);
    REQUIRE(read1 == fastq("read1", "", ""));
    REQUIRE(read2 == fastq("read2", "AA", "12"));
}


TEST_CASE("Extracting both sequences extending past each other", "[alignment::extract_adapter_sequences]")
{
    const fastq expected_1 = fastq("read1", "TTTTTTCCC", "ABCDEFGHI");
    const fastq expected_2 = fastq("read2", "AAGGGGGG",  "12345678");
    fastq read1 = expected_1;
    fastq read2 = expected_2;
    extract_adapter_sequences(ALN().offset(-2), read1, read2);
    REQUIRE(read1 == fastq("read1", "CCC", "GHI"));
    REQUIRE(read2 == fastq("read2", "AA", "12"));
}


///////////////////////////////////////////////////////////////////////////////
// Brute-force checking of alignment calculations
// Simply check all combinations involving 3 bases varying, for a range of
// sequence lengths to help catch corner cases with the optimizations

// The function is not exposed, so a declaration is required
bool compare_subsequences(const alignment_info& best, alignment_info& current,
                          const char* seq_1_ptr, const char* seq_2_ptr);


/** Naive reimplementation of alignment calculation. **/
void update_alignment(alignment_info& aln,
                      const std::string& a,
                      const std::string& b,
                      size_t nbases)
{
    // Don't count all these checks in test statistics
    if (a.length() != b.length()) {
       REQUIRE(a.length() == b.length());
    }

    for (size_t i = 0; i < nbases; ++i) {
        const char nt1 = a.at(i);
        const char nt2 = b.at(i);

        if (nt1 == 'N' || nt2 == 'N') {
            aln.n_ambiguous++;
        } else if (nt1 == nt2) {
            aln.score++;
        } else {
            aln.n_mismatches++;
            aln.score--;
        }
    }
}


/** Returns all 3 nt combinations of the bases ACGTN. **/
std::vector<std::string> get_combinations()
{
    std::vector<std::string> result;
    const std::string nts = "ACGTN";
    for (size_t i = 0; i < nts.length(); ++i) {
        for (size_t j = 0; j < nts.length(); ++j) {
            for (size_t k = 0; k < nts.length(); ++k) {
                std::string combination(3, 'A');
                combination.at(0) = nts.at(i);
                combination.at(1) = nts.at(j);
                combination.at(2) = nts.at(k);
                result.push_back(combination);
            }
        }
    }

    return result;
}


TEST_CASE("Brute-force validation", "[alignment::compare_subsequences]")
{
    const alignment_info best;
    const std::vector<std::string> combinations = get_combinations();
    for (size_t seqlen = 10; seqlen <= 20; ++seqlen) {
        for (size_t pos = 0; pos < seqlen; ++pos) {
            const size_t nbases = std::min<int>(3, seqlen - pos);

            for (size_t i = 0; i < combinations.size(); ++i) {
                for (size_t j = 0; j < combinations.size(); ++j) {
                    alignment_info expected;
                    expected.length = seqlen;
                    expected.score = seqlen - nbases;
                    update_alignment(expected, combinations.at(i), combinations.at(j), nbases);

                    std::string mate1 = std::string(seqlen, 'A');
                    mate1.replace(pos, nbases, combinations.at(i).substr(0, nbases));
                    std::string mate2 = std::string(seqlen, 'A');
                    mate2.replace(pos, nbases, combinations.at(j).substr(0, nbases));

                    alignment_info current;
                    current.length = seqlen;
                    compare_subsequences(best, current, mate1.c_str(), mate2.c_str());

                    // Don't count all these checks in test statistics
                    if (!(current == expected)) {
                        REQUIRE(current == expected);
                    }
                }
            }
        }
    }
}

} // namespace ar