1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237 238 239 240 241 242 243 244 245 246 247 248 249 250 251 252 253 254 255 256 257 258 259 260 261 262 263 264 265 266 267 268 269 270 271 272 273 274 275 276 277 278 279 280 281 282 283 284 285 286 287 288 289 290 291 292 293 294 295 296
//! Trims reads using 0-based coordinates
//!
//! # Examples
//!
//! ## Adapters
//!
//! ### Download the adapter files
//!
//! ```bash
//! mkdir -pv $HOME/db
//! pushd $HOME/db # step into the db directory
//! git clone https://github.com/lskatz/adapterseqs
//! ADAPTERS=$(find $HOME/db/adapterseqs -name '*.fa')
//! popd # return to the original directory
//! ```
//!
//! ### Trim the adapters
//!
//! ```bash
//! cat file.fastq | \
//! fasten_trim --adapterseqs <(echo -e ">test\nCTTT") > trimmed.fastq
//!
//! cat $HOME/db/adapterseqs/adapters/*.fa > ./adapters.fasta
//! cat file.fastq | \
//! fasten_trim --adapterseqs ./adapters.fasta > trimmed.fastq
//! ```
//!
//! ## Blunt-end trim five bases from the right side
//!
//! ```bash
//! cat file.fastq | fasten_trim -l -5 > trimmed.fastq
//! ```
//!
//! ## Keep a maximum of 100bp with blunt-end trimming on the right side
//!
//! ```bash
//! cat file.fastq | fasten_trim -l 99 > trimmed.fastq
//! ```
//!
//! ## Blunt-end trim 5bp from the left side
//!
//! ```bash
//! cat file.fastq | fasten_trim -f 4 > trimmed.fastq
//! ```
//!
//! # Usage
//!
//! ```text
//! Usage: fasten_trim [-h] [-n INT] [-p] [-v] [-f INT] [-l INT]
//!
//! Options:
//! -h, --help Print this help menu.
//! -n, --numcpus INT Number of CPUs (default: 1)
//! -p, --paired-end The input reads are interleaved paired-end
//! -v, --verbose Print more status messages
//! -f, --first-base INT
//! The first base to keep (default: 0)
//! -l, --last-base INT The last base to keep. (default: 0)
//! -a, --adapterseqs path/to/file.fa
//! fasta file of adapters
//! ```
//!
//! # Notes
//!
//! The algorithm is as follows:
//!
//! 1. marks the first and last bases for trimming as 0 and the last base, respectively
//! 2. if an adapter is found at the beginning of the sequence, then move the marker for where it will be trimmed
//! 3. Compare the blunt end suggested trimming against where an adapter might be found and move the marker as the most inward possible
//! 4. Trim the sequence and quality strings
//!
//! Making the output more explicit while combining both algorithms can involve a two step process:
//!
//! ```bash
//! cat file.fastq | \
//! fasten_trim --adapterseqs ./adapters.fasta | \
//! fasten_trim -f 4 -l 99 > trimmed.fastq
//! ```
//!
//! # Output
//!
//! The deflines will be altered with a description of the trimming using key=value syntax, separated by spaces, e.g.,
//! `@M03235:53:000000000-AHLTD:1:1101:1826:14428 trimmed_adapter_rev=TT trimmed_left=0 trimmed_right=249`
//! or for a forward adapter,
//! `@M03235:53:000000000-AHLTD:1:1101:1758:14922 trimmed_adapter_fwd=AA trimmed_left=2 trimmed_right=251`
extern crate fasten;
extern crate statistical;
extern crate getopts;
extern crate threadpool;
use std::fs::File;
use std::io::BufReader;
use std::cmp::{min,max};
use std::process::exit;
use std::collections::HashMap;
use std::io::BufRead;
use fasten::fasten_base_options;
use fasten::fasten_base_options_matches;
use fasten::logmsg;
use fasten::reverse_complement;
use fasten::io::fastq;
use fasten::io::seq::Seq;
fn main(){
let mut opts = fasten_base_options();
// script-specific options
opts.optopt("f","first-base","The first base to keep (default: 0)","INT");
opts.optopt("l","last-base","The last base to keep (default: 0)","INT");
opts.optopt("a","adapterseqs","fasta file of adapters","path/to/file.fa");
let matches = fasten_base_options_matches("Blunt-end trims using 0-based coordinates", opts);
let adapterseqs:String = {
if matches.opt_present("adapterseqs") {
matches.opt_str("adapterseqs")
.expect("ERROR: could not understand parameter --adapterseqs")
} else {
"".to_string()
}
};
// store the adapter sequences as a vector of strings
let mut adapters:Vec<String> = Vec::new();
if matches.opt_present("adapterseqs") && adapterseqs.len() > 0 {
// check that the file path exists
// if not, exit with an error
if !std::path::Path::new(&adapterseqs).exists() {
logmsg(format!("ERROR: adapter file {} does not exist", &adapterseqs));
exit(1);
}
// read the adapter sequences from the fasta file
adapters = read_fasta(&adapterseqs)
.values()
.map(|x| x.to_string())
.collect();
}
//if matches.opt_present("verbose") {
// //logmsg(&adapters);
// eprintln!("Adapters: {:?}", adapters);
// exit(3);
//}
let first_base:usize ={
if matches.opt_present("first-base") {
matches.opt_str("first-base")
.expect("ERROR: could not understand parameter --first-base")
.parse()
.expect("ERROR: --first-base is not an INT")
} else {
0
}
};
let last_base:usize ={
if matches.opt_present("last-base") {
matches.opt_str("last-base")
.expect("ERROR: could not understand parameter --last-base")
.parse()
.expect("ERROR: --last-base is not an INT")
} else {
0
}
};
let _num_cpus:usize = {
if matches.opt_present("numcpus") {
/*
matches.opt_str("numcpus")
.expect("ERROR: could not understand parameter --numcpus")
.parse()
.expect("ERROR: --numcpus is not an INT");
*/
logmsg("Warning: multithreading this script currently slows it down. Resetting to 1 cpu. Avoid this warning by not using --numcpus");
1 as usize
} else {
1 as usize
}
};
// Read from stdin
let my_file = File::open("/dev/stdin").expect("Could not open file");
let my_buffer=BufReader::new(my_file);
let fastq_reader = fastq::FastqReader::new(my_buffer);
let fastq_iter = fastq_reader.into_iter();
for seq in fastq_iter {
let trimmed:String = trim_worker(seq, first_base, last_base, &adapters);
println!("{}", trimmed);
}
}
/// Trim a set of fastq entries and send it to a channel
fn trim_worker(seq:Seq, suggested_first_base:usize, suggested_last_base:usize, adapters:&Vec<String> ) -> String {
// In this function, keep track of where the first and
// last base would be trimmed with a simple marker.
// Most instances of the word "trimming" in this function is just moving first_base and last_base.
let mut first_base = 0;
// The last position is either the last_base parameter
// or the last position in the string, whichever is less.
let mut last_base = seq.seq.len()-1;
// Make note of what is trimmed
let mut description = String::new();
// First, run the adapter trimming, before any blunt end trimming
// First, detect if there are any adapters in the sequence
// If there are, then trim the sequence at the adapter
for adapter in adapters {
let adapter_length = adapter.len();
// If the adapter is longer than the sequence, skip it: it won't exist in the sequence as a whole adapter.
if adapter_length >= seq.seq.len() {
continue;
}
// Check if the adapter is at the beginning of the sequence
if &seq.seq[0..adapter_length] == adapter {
first_base = adapter_length;
description.push_str(&format!(" trimmed_adapter_fwd={}", &adapter));
}
// Check if the revcom is at the end of the sequence
let revcom = reverse_complement(&adapter);
let end_slice: &str = &seq.seq[&seq.seq.len()-1 - adapter_length..].trim();
if end_slice == revcom {
last_base = seq.seq.len() - adapter_length;
description.push_str(&format!(" trimmed_adapter_rev={}", &revcom));
}
}
// Next, run the blunt end trimming.
// Take the maximum between the suggested left trim and the current left trim.
// If the left trim is longer than the sequence length, then omit a warning and do not trim.
first_base = max(first_base, suggested_first_base);
if first_base >= seq.seq.len() {
logmsg("Warning: the left trim is longer than the sequence length. Skipping.");
first_base = 0;
}
// Take the minimum between the suggested right trim and the current right trim.
// If the last base is less than 1, then omit a warning and do not trim.
last_base = {
if suggested_last_base == 0 {
last_base
} else {
min(last_base, suggested_last_base)
}
};
if last_base < 1 {
logmsg("Warning: the right trim is longer than the sequence length. Skipping.");
last_base = seq.seq.len()-1;
}
description.push_str(&format!(" trimmed_left={} trimmed_right={}", first_base, last_base-1));
let sequence = &seq.seq[first_base..last_base];
let quality = &seq.qual[first_base..last_base];
let trimmed = format!("{}{}\n{}\n+\n{}", seq.id, description, sequence, quality);
return trimmed;
}
// Taken from https://medium.com/bioinformatics-with-rust/how-to-read-a-fasta-file-9472b77589f7
/// Read a fasta file and return a HashMap of the sequences
fn read_fasta(file_path: &str) -> HashMap<String, String> {
let mut data = HashMap::new();
let file = File::open(file_path).expect("Invalid filepath");
let reader = BufReader::new(file);
let mut seq_id = String::new();
for line in reader.lines() {
let line = line.unwrap();
// Check if the line starts with '>' (indicating a sequence ID or header)
if line.starts_with('>') {
seq_id = line.trim_start_matches('>').to_string();
} else {
// If it's a DNA sequence line, insert or update the HashMap entry
// If seq_id is not present, insert a new entry with an empty String
// Then append the current line to the existing DNA sequence
data.entry(seq_id.clone()).or_insert_with(String::new).push_str(&line);
}
}
data
}