Structural Variant Detection

Sniffles2 - Basic SV Calling

# Call SVs from aligned BAM
sniffles --input aligned.bam \
    --vcf structural_variants.vcf \
    --reference reference.fa \
    --threads 4

Sniffles2 - Common Options

sniffles --input aligned.bam \
    --vcf structural_variants.vcf \
    --reference reference.fa \
    --threads 8 \
    --minsupport 3 \               # Min supporting reads
    --minsvlen 50 \                # Min SV length
    --mapq 20 \                    # Min mapping quality
    --output-rnames \              # Include read names
    --mosaic                       # Detect mosaic SVs

Sniffles2 - Population Calling

# Step 1: Call SVs per sample with SNF output
sniffles --input sample1.bam --snf sample1.snf --reference reference.fa
sniffles --input sample2.bam --snf sample2.snf --reference reference.fa

# Step 2: Merge and genotype
sniffles --input sample1.snf sample2.snf \
    --vcf population_svs.vcf \
    --reference reference.fa

cuteSV - Alternative Caller

# cuteSV SV calling
cuteSV aligned.bam reference.fa output.vcf work_dir/ \
    --threads 8 \
    --min_support 3 \
    --min_size 50 \
    --genotype

cuteSV - ONT Optimized

# Settings optimized for ONT
cuteSV aligned.bam reference.fa output.vcf work_dir/ \
    --threads 8 \
    --max_cluster_bias_INS 100 \
    --diff_ratio_merging_INS 0.3 \
    --max_cluster_bias_DEL 100 \
    --diff_ratio_merging_DEL 0.3 \
    --genotype

cuteSV - PacBio HiFi Optimized

# Settings optimized for HiFi
cuteSV aligned.bam reference.fa output.vcf work_dir/ \
    --threads 8 \
    --max_cluster_bias_INS 1000 \
    --diff_ratio_merging_INS 0.9 \
    --max_cluster_bias_DEL 1000 \
    --diff_ratio_merging_DEL 0.5 \
    --genotype

SVIM - Another Alternative

# SVIM for ONT data
svim alignment output_dir/ aligned.bam reference.fa \
    --insertion_sequences \
    --read_names \
    --sample sample_name

pbsv - PacBio Specific

# Discover signatures
pbsv discover aligned.bam signatures.svsig.gz

# Call SVs
pbsv call reference.fa signatures.svsig.gz structural_variants.vcf

Filter SV Calls

# Filter by quality and size
bcftools filter -i 'QUAL>=20 && ABS(SVLEN)>=50' svs.vcf > svs.filtered.vcf

# Keep only PASS
bcftools view -f PASS svs.vcf > svs.pass.vcf

# Filter specific SV types
bcftools view -i 'SVTYPE="DEL"' svs.vcf > deletions.vcf
bcftools view -i 'SVTYPE="INS"' svs.vcf > insertions.vcf

Merge Multiple Callers

# Use SURVIVOR to merge SV callsets
SURVIVOR merge sample_files.txt 1000 2 1 1 0 50 merged_svs.vcf

# sample_files.txt contains VCF paths, one per line
# Parameters: max_distance, min_callers, type_agree, strand_agree, est_distance, min_size

Annotate SVs

# Annotate with AnnotSV
AnnotSV -SVinputFile svs.vcf \
    -genomeBuild GRCh38 \
    -outputFile annotated_svs

# Or with bcftools
bcftools annotate -a gnomad_sv.vcf.gz -c INFO svs.vcf > svs.annotated.vcf

SV Types

Key Parameters - Sniffles2

Key Parameters - cuteSV

Coverage Guidelines

Related Skills

• long-read-alignment - Generate input BAM
• medaka-polishing - Polish assembly with SVs
• variant-calling/structural-variant-calling - Short-read SV comparison