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Updated: Jan 24, 2026

Separation and Fractionation of Culture Filtrate Proteins (CFPs) from Mycobacterium tuberculosis
Published on: July 11, 2025
Whole genome sequencing Mycobacterium tuberculosis directly from sputum identifies more genetic diversity than
Camus Nimmo1,2, Liam P Shaw3,4, Ronan Doyle5,6
1Division of Infection and Immunity, University College London, London, WC1E 6BT, UK. c.nimmo.04@cantab.net.
Background:
Repeated culture reduces within-sample Mycobacterium tuberculosis genetic diversity due to selection of clones suited to growth in culture and/or random loss of lineages, but it is not known to what extent omitting the culture step altogether alters genetic diversity. We compared M. tuberculosis whole genome sequences generated from 33 paired clinical samples using two methods. In one method DNA was extracted directly from sputum then enriched with custom-designed SureSelect (Agilent) oligonucleotide baits and in the other it was extracted from mycobacterial growth indicator tube (MGIT) culture.
Results:
DNA directly sequenced from sputum showed significantly more within-sample diversity than that from MGIT culture (median 5.0 vs 4.5 heterozygous alleles per sample, p = 0.04). Resistance associated variants present as HAs occurred in four patients, and in two cases may provide a genotypic explanation for phenotypic resistance.
Conclusions:
Culture-free M. tuberculosis whole genome sequencing detects more within-sample diversity than a leading culture-based method and may allow detection of mycobacteria that are not actively replicating.
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