nanoMLST

nanoMLST performs multilocus sequence typing (MLST) using Oxford Nanopore Technologies MinION sequencing with dual-barcode multiplexing to assign MLST alleles for microbial strain typing and lineage analysis.

Key Features:

• Dual-Barcoding Multiplexing: Employs a dual-barcode system to process multiple samples concurrently on a single MinION flow cell, enabling multiplexed sequencing.
• Sequencing Efficiency: Sequences seven full-length housekeeping genes from up to 96 meticillin-resistant Staphylococcus aureus isolates in a single run, supporting high-throughput MLST.
• High Accuracy: Demonstrates 100% agreement in allele assignments when compared to conventional Sanger sequencing.
• Scalability: Estimated to handle MLST for up to 1,000 samples simultaneously for large-scale genotyping projects.
• Speed and Cost-Effectiveness: Provides a rapid and lower-cost alternative to Sanger sequencing for MLST using the MinION platform.

Scientific Applications:

• Microbial lineage and strain typing: Enables MLST-based strain typing and lineage determination of bacterial isolates.
• Epidemiological surveillance: Supports epidemiological studies and surveillance by enabling genotyping of large sample sets.
• Outbreak investigation: Facilitates outbreak investigations through rapid MLST allele assignment across many isolates.
• Antimicrobial resistance research: Assists research into antimicrobial resistance patterns by providing MLST data for resistant strains such as meticillin-resistant Staphylococcus aureus.

Methodology:

Resulting MinION sequencing data undergo bioinformatic analysis to assign MLST alleles.