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Comparative analysis of the sensitivity of metagenomic sequencing and PCR to detect a biowarfare simulant ($Bacillus\ atrophaeus$) in soil samples

Abstract : To evaluate the sensitivity of high-throughput DNA sequencing for monitoring biowarfare agents in the environment, we analysed soil samples inoculated with different amounts of $Bacillus\ atrophaeus$, a surrogate organism for $Bacillus\ anthracis$. The soil samples considered were a poorly carbonated soil of the silty sand class, and a highly carbonated soil of the silt class. Control soil samples and soil samples inoculated with 10, 10$^3$ , or 10$^5$ cfu were processed for DNA extraction. About 1% of the DNA extracts was analysed through the sequencing of more than 108 reads. Similar amounts of extracts were also studied for $Bacillus\ atrophaeus$ DNA content by real-time PCR. We demonstrate that, for both soils, highthroughput sequencing is at least equally sensitive than real-time PCR to detect $Bacillus\ atrophaeus$ DNA. We conclude that metagenomics allows the detection of less than 10 ppm of DNA from a biowarfare simulant in complex environmental samples.
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https://hal-cea.archives-ouvertes.fr/cea-03282656
Contributor : Marie-Claude Marsolier <>
Submitted on : Tuesday, July 13, 2021 - 11:21:49 AM
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Delphine Plaire, Simon Puaud, Marie-Claude Marsolier-Kergoat, Jean-Marc Elalouf. Comparative analysis of the sensitivity of metagenomic sequencing and PCR to detect a biowarfare simulant ($Bacillus\ atrophaeus$) in soil samples. PLoS ONE, Public Library of Science, 2017, 12 (5), pp.e0177112. ⟨10.1371/journal.pone.0177112⟩. ⟨cea-03282656⟩

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