QIAseq xHYB Mycobacterium tuberculosis Panel

• Enables whole genome sequencing of Mycobacterium tuberculosis (MTB) without the need for bacterial culture 
• Covers all TB antimicrobial resistance (AMR) markers 
• Designed against the seven major lineages of MTB for comprehensive analysis 
• Uses hybrid capture enrichment and QIAseq xHYB Chemistry  

QIAseq xHYB Mycobacterium tuberculosis Panel allows for the comprehensive culture-free whole genome sequencing (WGS) of tuberculosis from samples such as sputum or cerebrospinal fluid (CSF), streamlining the process and facilitating rapid detection and characterization of MTB strains. 

Thanks to targeted whole genome enrichment (WGE), the QIAseq xHYB Mycobacterium tuberculosis Panel gives high-level coverage of the whole MTB genome, even from lower-quality samples ( Figure 1,  Figure 2,  Figure 3). 

Benefits of WGE over targeted-amplicon approaches 

• Comprehensive coverage: WGE enables the detection of mutations and variations across the entire MTB genome, including non-coding regions and regulatory elements. 
• Enhanced antimicrobial resistance detection: WGE detects novel antimicrobial resistance markers and provides a comprehensive assessment of genetic determinants of drug resistance. 
• Lineage identification: WGE accurately determines the lineage and phylogenetic relationship of MTB strains, offering valuable insights into MTB transmission dynamics and epidemiology. 
• Future-proofing: WGE allows for retrospective analysis of past samples. Furthermore, if you need to reanalyze the samples in the future, samples prepared with WGE can be reanalyzed for newly discovered genetic markers and drug resistance mechanisms. 

The QIAseq xHYB Mycobacterium tuberculosis Panel uses targeted WGE, in which hybrid capture probes are tiled across the entire MTB genome. The panel is designed against multiple representative sequences from each of the seven major MTB lineages, allowing for full coverage of MTB diversity. The panel covers all known resistance-related genes.

QIAseq FX DNA library construction 

Both isolated genomic DNA or total nucleic acid can be converted to Illumina-compatible NGS libraries using the QIAseq FX DNA Library Kit. DNA products are enzymatically sheared, the fragmented DNA is end-repaired, and an “A” is added to the 3’-end. Illumina-platform-specific adapters are ligated. The adapters contain the necessary sequences for library binding during flow-cell sequencing. After purification, libraries are amplified to generate sufficient yields for the hybrid capture reaction. 

Hybrid capture 

Pre-capture libraries are hybridized to the QIAseq xHYB Mycobacterium tuberculosis probes. Probe-target hybrids are then bound to streptavidin-coated magnetic beads and washed. Enriched libraries are then amplified and prepared for sequencing on Illumina systems. 

Analysis

Resulting FASTQ can be analyzed using QIAGEN CLC Genomics Workbench or with any method compatible with MTB WGS sequencing. CLC Genomic Workbench provides a comprehensive report on the MTB lineage designation, spoligotyping and resistance associated genes.

QIAseq xHYB Mycobacterium tuberculosis is ideal for clinical laboratories, public health agencies and research institutions seeking a robust panel for MTB surveillance and research.   

This panel can be used for: 

• MTB lineage assignment 
• Resistance typing 
• Real-time outbreak surveillance