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use super::*;
use bio::alphabets::dna::revcomp;
use lazy_static::lazy_static;
use rayon::{current_num_threads, prelude::*};
use regex::Regex;
use rust_htslib::{bam, bam::record::Aux, bam::Read};
use std::convert::TryFrom;
use std::time::Instant;
pub struct BaseMods {
pub modified_base: u8,
pub strand: char,
pub modification_type: char,
pub modified_positions: Vec<i64>,
pub modified_reference_positions: Vec<i64>,
}
impl BaseMods {
pub fn new(record: &bam::Record) -> Vec<BaseMods> {
lazy_static! {
static ref MM_RE: Regex =
Regex::new(r"((([ACGTUN])([-+])([a-z]+|[0-9]+))[.?]?((,[0-9]+)*;)*)").unwrap();
}
let mut rtn = vec![];
if let Ok(Aux::String(mm_text)) = record.aux(b"MM") {
for cap in MM_RE.captures_iter(mm_text) {
let mod_base = cap.get(3).map(|m| m.as_str().as_bytes()[0]).unwrap();
let mod_strand = cap.get(4).map_or("", |m| m.as_str());
let modification_type = cap.get(5).map_or("", |m| m.as_str());
let mod_dists_str = cap.get(6).map_or("", |m| m.as_str());
let mod_dists: Vec<i64> = mod_dists_str
.trim_end_matches(';')
.split(',')
.map(|s| s.trim())
.filter(|s| !s.is_empty())
.map(|s| s.parse().unwrap())
.collect();
let forward_bases = if record.is_reverse() {
revcomp(record.seq().as_bytes())
} else {
record.seq().as_bytes()
};
let mut cur_mod_idx = 0;
let mut cur_seq_idx = 0;
let mut dist_from_last_mod_base = 0;
let mut modified_positions: Vec<i64> = vec![0; mod_dists.len()];
while cur_seq_idx < forward_bases.len() && cur_mod_idx < mod_dists.len() {
let cur_base = forward_bases[cur_seq_idx];
if cur_base == mod_base && dist_from_last_mod_base == mod_dists[cur_mod_idx] {
modified_positions[cur_mod_idx] = i64::try_from(cur_seq_idx).unwrap();
dist_from_last_mod_base = 0;
cur_mod_idx += 1;
} else if cur_base == mod_base {
dist_from_last_mod_base += 1
}
cur_seq_idx += 1;
}
assert_eq!(cur_mod_idx, mod_dists.len());
let mut mods = BaseMods {
modified_base: mod_base,
strand: mod_strand.chars().next().unwrap(),
modification_type: modification_type.chars().next().unwrap(),
modified_positions,
modified_reference_positions: vec![],
};
mods.add_reference_positions(record);
rtn.push(mods);
}
} else {
log::debug!("No MM tag found");
}
rtn
}
pub fn add_reference_positions(&mut self, record: &bam::Record) {
let positions = positions_on_complimented_sequence(record, &self.modified_positions);
self.modified_reference_positions = liftover_exact(record, &positions);
}
}
pub fn merge_two_lists<T>(left: Vec<T>, right: Vec<T>) -> Vec<T>
where
T: Ord,
T: Clone,
{
let mut x = [left, right].concat();
x.sort();
x
}
pub fn get_u32_tag(record: &bam::Record, tag: &[u8; 2]) -> Vec<i64> {
if let Ok(Aux::ArrayU32(array)) = record.aux(tag) {
let read_array = array.iter().map(|x| x as i64).collect::<Vec<_>>();
read_array
} else {
vec![]
}
}
pub struct FiberseqData {
pub record: bam::Record,
pub nuc_starts: Vec<i64>,
pub msp_starts: Vec<i64>,
pub nuc_length: Vec<i64>,
pub msp_length: Vec<i64>,
pub ref_nuc: Vec<(i64, i64)>,
pub ref_msp: Vec<(i64, i64)>,
pub base_mods: Vec<BaseMods>,
}
impl FiberseqData {
pub fn new(record: &bam::Record) -> Self {
let nuc_starts = get_u32_tag(record, b"ns");
let msp_starts = get_u32_tag(record, b"as");
let nuc_length = get_u32_tag(record, b"nl");
let msp_length = get_u32_tag(record, b"al");
let ref_nuc = get_closest_reference_range(&nuc_starts, &nuc_length, record);
let ref_msp = get_closest_reference_range(&msp_starts, &msp_length, record);
FiberseqData {
record: record.clone(),
nuc_starts,
msp_starts,
nuc_length,
msp_length,
ref_nuc,
ref_msp,
base_mods: BaseMods::new(record),
}
}
pub fn from_records(records: &Vec<bam::Record>) -> Vec<Self> {
records
.par_iter()
.map(FiberseqData::new)
.collect::<Vec<_>>()
}
pub fn to_string(&mut self, reference: bool, starts: &[i64], lengths: &[i64]) -> String {
let ct;
let start;
let end;
let name = std::str::from_utf8(self.record.qname()).unwrap();
if reference {
ct = "";
start = self.record.reference_start();
end = self.record.reference_end();
} else {
ct = name;
start = 0;
end = self.record.seq_len() as i64;
}
let strand = self.record.is_reverse();
let score = 0;
let color = "126,126,126";
let b_ct = starts.len();
let b_st: String = starts.iter().map(|&id| id.to_string() + ",").collect();
let b_ln: String = lengths.iter().map(|&id| id.to_string() + ",").collect();
format!("{ct}\t{start}\t{end}\t{name}\t{score}\t{strand}\t{start}\t{end}\t{color}\t{b_ct}\t{b_ln}\t{b_st}\n")
}
}
pub fn process_bam_chunk(
records: &Vec<bam::Record>,
so_far: usize,
_reference: bool,
m6a: &Option<String>,
_cpg: &Option<String>,
_msp: &Option<String>,
_nuc: &Option<String>,
) {
let start = Instant::now();
let _fiber_data = FiberseqData::from_records(records);
let duration = start.elapsed().as_secs_f64();
log::info!(
"Processing {:.2} reads/second. {:} reads done so far.",
records.len() as f64 / duration,
so_far + records.len()
);
match m6a {
Some(m6a) => {
log::info!("Processing {:}", m6a);
}
None => {}
}
}
pub fn extract_contained(
bam: &mut bam::Reader,
reference: bool,
m6a: &Option<String>,
cpg: &Option<String>,
msp: &Option<String>,
nuc: &Option<String>,
) {
let bin_size = current_num_threads() * 2000;
let mut cur_count = 0;
let mut cur_vec = vec![];
let mut proccesed_reads = 0;
for r in bam.records() {
let record = r.unwrap();
cur_vec.push(record);
cur_count += 1;
if cur_count == bin_size {
process_bam_chunk(&cur_vec, proccesed_reads, reference, m6a, cpg, msp, nuc);
proccesed_reads += cur_vec.len();
cur_vec.clear();
cur_count = 0;
}
}
process_bam_chunk(&cur_vec, proccesed_reads, reference, m6a, cpg, msp, nuc);
}