P011
Culture-Free Targeted Whole Genome Enrichment of Mycobacterium tuberculosis and Next-Generation Sequencing Enables Rapid Surveillance of Drug Resistance and Genomic Tracking.
M Fosbrink(1) M ZAIS(1) R Larsen(1) C Andersen(1) L Schauser(1) D Barbera(1) J Shaffer(1)
1:QIAGEN Sciences
Whole-Genome Enrichment (WGE) of Mycobacterium tuberculosis (MTB), using hybrid capture technology, offers a powerful approach to elucidate the genetic diversity, evolution and drug resistance of MTB using Next-Generation Sequencing. By enriching the complete genome, WGE overcomes challenges of MTB characterization such as long culture times and low bacterial DNA content in samples while enabling high-resolution analysis of key loci associated with lineage identification, drug resistance, and strain differentiation.
QIAGEN’s QIAseq Mycobacterium tuberculosis Panel is a hybrid-capture based approach that enriches the complete genome to enable NGS characterization. In this study, we used this product to demonstrate its utility for rapid whole genome analysis. The NGS results suggest the panel covered 93.04% of the genome at 10x base coverage for 300 genome copies of H37Rv that was spiked into 100 ng of sputum DNA. Increasing the input amount to 10,000 genome copies exhibited 95.22% of the genome covered at 100x. Using the dedicated Mycobacterium tuberculosis workflow from CLC Microbial Genomics Module, the correct resistance calls with corresponding mutations were determined for strains ATCC 35838 and ATCC 35822. Additionally, the panel was able to identify accessory genes found in H37Ra that are not in the reference sequence of H37Rv.
QIAGEN’s QIAseq Mycobacterium tuberculosis Panel is a powerful genomic sequencing tool that allows for rapid whole genome sequencing of DNA that was extracted directly from a sample with no culture requirements. Data analysis using CLC Microbial Genomics Module allows for identification of mutations that contribute to drug resistance and variants for tracking transmission.