好久不更新,上个硬货。脚本需提前准备宿主和rrna的bowtie2索引文件,原始数据的命名方式为{sample}_raw_1/2.fq.gz,保存有原始数据路径的文件,保存样品列表的文件。
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#!/bin/bash
# 脚本功能:二代测序数据质控、去rRNA与去宿主序列处理
#版本1.2,新增了--rrna功能
usage() {
echo "用法: $0 [--host <宿主类型>] [--rrna] <路径文件> <样品名列表文件>"
exit 1
}
# ---------- 配置路径 ----------
declare -A HOST_INDEX=(
[sheep]="/mnt/data/database/bowtie/sheep/sheepgenome"
[goat]="/mnt/data/database/bowtie/goat/goatgenome"
[cattle]="/mnt/data/database/bowtie/cattle/cattlegenome"
)
SSU_INDEX="/mnt/data/database/bowtie/SSU/SSU"
LSU_INDEX="/mnt/data/database/bowtie/LSU/LSU"
# 初始化变量
HOST=""
REMOVE_RRNA=false
INPUT_FILE=""
SAMPLE_LIST=""
# 增强参数解析
while [[ $# -gt 0 ]]; do
case "$1" in
--host)
# 宿主类型验证
if [[ ! " sheep goat cattle " =~ " $2 " ]]; then
echo "错误:无效宿主类型 '$2',可选: sheep/goat/cattle"
exit 1
fi
HOST="$2"
shift 2 ;;
--rrna)
REMOVE_RRNA=true
shift ;;
--help)
usage ;;
-*)
echo "错误:未知选项 $1"
usage ;;
*)
# 位置参数处理
if [[ -z $INPUT_FILE ]]; then
INPUT_FILE="$1"
elif [[ -z $SAMPLE_LIST ]]; then
SAMPLE_LIST="$1"
else
echo "错误:多余参数 $1"
usage
fi
shift ;;
esac
done
# 验证必要参数
if [[ -z $INPUT_FILE || -z $SAMPLE_LIST ]]; then
echo "错误:缺失必要参数!"
usage
fi
# 读取原始路径并验证
INPUT_DIR=$(cat "$INPUT_FILE")
if [[ ! -d $INPUT_DIR ]]; then
echo "错误:路径文件中的目录 $INPUT_DIR 不存在"
exit 1
fi
# 宿主索引验证
if [[ -n $HOST ]]; then
INDEX="${HOST_INDEX[$HOST]}"
if [[ ! -f "${INDEX}.1.bt2" ]]; then
echo "错误:宿主索引文件 ${INDEX}.1.bt2 不存在"
exit 1
fi
fi
# ---------- 目录创建 ----------
CLEAN_DIR=$(dirname "$INPUT_DIR")/cleandata
mkdir -p "$CLEAN_DIR" || { echo "错误:无法创建目录 $CLEAN_DIR"; exit 1; }
# ---------- 处理流程 ----------
process_sample() {
local sample=$1
local raw_1="${INPUT_DIR}/${sample}_raw_1.fq.gz"
local raw_2="${INPUT_DIR}/${sample}_raw_2.fq.gz"
local clip_1="${CLEAN_DIR}/${sample}.clip.1.fq.gz"
local clip_2="${CLEAN_DIR}/${sample}.clip.2.fq.gz"
# Step 1: 质控(fastp默认参数)
echo "[$(date)] 处理样品 $sample:质控中..."
fastp -i "$raw_1" -I "$raw_2" -o "$clip_1" -O "$clip_2" || {
echo "错误:fastp处理失败!"
exit 1
}
#Step 2: 去宿主
if [[ -n $HOST ]]; then
echo "[$(date)] 去除宿主 $HOST 序列..."
bowtie2 -p 192 -x "$INDEX" --quiet\
-1 "$clip_1" -2 "$clip_2" \
--un-conc-gz "${CLEAN_DIR}/${sample}.clean.fq.gz" \
--al-conc-gz "${CLEAN_DIR}/${sample}.host.fq.gz" \
-S /dev/null > "${CLEAN_DIR}/${sample}.bowtie2.log" || exit 1
mv "${CLEAN_DIR}/${sample}.clean.fq.1.gz" "$clip_1"
mv "${CLEAN_DIR}/${sample}.clean.fq.2.gz" "$clip_2"
fi
#Step 3: 去rRNA
if $REMOVE_RRNA; then
echo "[$(date)] 去除rRNA序列(SSU+LSU)..."
# SSU过滤
bowtie2 -p 192 -x "$SSU_INDEX" --quiet\
-1 "$clip_1" -2 "$clip_2" \
--un-conc-gz "${CLEAN_DIR}/${sample}.clean.fq.gz" \
--al-conc-gz "${CLEAN_DIR}/${sample}.SSU.fq.gz" \
-S /dev/null > "${CLEAN_DIR}/${sample}.SSU.log" || exit 1
mv "${CLEAN_DIR}/${sample}.clean.fq.1.gz" "$clip_1"
mv "${CLEAN_DIR}/${sample}.clean.fq.2.gz" "$clip_2"
# LSU过滤
bowtie2 -p 192 -x "$LSU_INDEX" --quiet\
-1 "$clip_1" -2 "$clip_2" \
--un-conc-gz "${CLEAN_DIR}/${sample}.clean.fq.gz" \
--al-conc-gz "${CLEAN_DIR}/${sample}.LSU.fq.gz" \
-S /dev/null > "${CLEAN_DIR}/${sample}.LSU.log" || exit 1
mv "${CLEAN_DIR}/${sample}.clean.fq.1.gz" "$clip_1"
mv "${CLEAN_DIR}/${sample}.clean.fq.2.gz" "$clip_2"
fi
}
# 遍历样品列表
while IFS= read -r sample; do
[[ -z $sample ]] && continue # 跳过空行
process_sample "$sample"
done < "$SAMPLE_LIST"
mkdir ${CLEAN_DIR}/host ${CLEAN_DIR}/rRNA ${CLEAN_DIR}/log
mv ${CLEAN_DIR}/*host\.fq* ${CLEAN_DIR}/host/
mv ${CLEAN_DIR}/*SU\.fq* ${CLEAN_DIR}/rRNA/
mv ${CLEAN_DIR}/*log ${CLEAN_DIR}/log
echo "[$(date)] 处理完成!结果保存至:$CLEAN_DIR"