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Mycobacterium tuberculosis complex CRISPR genotyping: improving efficiency, throughput and discriminative power of 'spoligotyping' with new spacers and a microbead-based hybridization assay.

Abstract : The aims of the present study were to implement a microbead-based 'spoligotyping' technique and to evaluate improvements by the addition of a panel of 25 extra spacers that we expected to provide an increased resolution on principal genetic group 1 (PGG 1) strains. We confirmed the high sensitivity and reproducibility of the classical technique using the 43 spacer panel and we obtained perfect agreement between the membrane-based and the microbead-based techniques. We further demonstrated an increase in the discriminative power of an extended 68 spacer format for differentiation of PGG 1 clinical isolates, in particular for the East African-Indian clade. Finally, we define a limited yet highly informative reduced 10 spacer panel set which could offer a more cost-effective option for implementation in resource-limited countries and that could decrease the need for additional VNTR (variable number of tandem repeats) genotyping work in molecular epidemiological studies. We also present an economic analysis comparing membrane-based and microbead-based techniques.
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https://hal.archives-ouvertes.fr/hal-00527223
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Submitted on : Monday, October 18, 2010 - 3:07:07 PM
Last modification on : Saturday, June 25, 2022 - 10:01:53 PM

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Jian Zhang, Edgar Abadia, Guislaine Refregier, Silva Tafaj, Maria Laura Boschiroli, et al.. Mycobacterium tuberculosis complex CRISPR genotyping: improving efficiency, throughput and discriminative power of 'spoligotyping' with new spacers and a microbead-based hybridization assay.. Journal of Medical Microbiology, Society for General Microbiology, 2010, 59 (Pt 3), pp.285-94. ⟨10.1099/jmm.0.016949-0⟩. ⟨hal-00527223⟩

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