ShahSanjivkumar_HS.402.1.1.14 _Data-Metadata_Development of HTM_Manuscript-20250402

Since the national validation of the sponge-stick based method for detection of Bacillus anthracis spores in environmental samples, there have not been focused efforts to address the low throughput nature of the method, which processes only one sample at one time. Sample processing remains a serious bottleneck for rapidly analyzing large numbers of samples expected from a biological warfare attack. Therefore, we developed a high-throughput method to simultaneously process multiple sponge-stick samples to be better prepared for rapid response and recovery after wide area anthrax incidents. In this method, sponges are placed in 50 mL tubes containing 25 mL buffer and shaken to release spores, after which the buffer is recovered for analysis. We determined that an additional rinse step, conducted in the same tubes with 10 mL buffer, further increased spore recovery from sponge-stick by approximately 10%. We determined that orbital shaking and multi-tube vortexing were both more effective than reciprocating shaking for recovering spores. We conducted simultaneous processing of up to 12 sponge-stick samples and demonstrated comparable spore recovery efficiencies to the traditional low-throughput stomacher-based method (approximately 60% recovery at 102-spore level and 75% recovery at 104-spore level for both methods in three replicate experiments, P > 0.05 for two-tailed T-tests for each experiment and spore level). We also demonstrated that our high-throughput method could be integrated with Rapid Viability-Polymerase Chain Reaction (RV-PCR) analysis and could detect levels as low as 40 spores per sponge even when challenged by a PCR particulate contaminant. This dataset is associated with the following publication: Brisson, V., S. Kane, and S. Shah. Development of a high-throughput method for processing sponge-stick samples to detect viable Bacillus anthracis spores. JOURNAL OF MICROBIOLOGICAL METHODS. Elsevier Science Ltd, New York, NY, USA, 235: 107149, (2025).

                                            [
  "020:00"
]
                                        

                                            {
  "fn": "Sanjivkumar Shah",
  "hasEmail": "mailto:shah.sanjiv@epa.gov"
}
                                        

                                            [
  {
    "title": "Manuscript_Devlopment of HT Method for Processing-B anthracis Spores-20250401.docx",
    "mediaType": "application/vnd.openxmlformats-officedocument.wordprocessingml.document",
    "downloadURL": "https://pasteur.epa.gov/uploads/10.23719/1532127/Manuscript_Devlopment%20of%20HT%20Method%20for%20Processing-B%20anthracis%20Spores-20250401.docx"
  },
  {
    "title": "ShahSanjivkumar_HS.402.1.1.14 _Data-Metadata_Development of HTM_Manuscript-20250402.docx",
    "mediaType": "application/vnd.openxmlformats-officedocument.wordprocessingml.document",
    "downloadURL": "https://pasteur.epa.gov/uploads/10.23719/1532127/ShahSanjivkumar_HS.402.1.1.14%20_Data-Metadata_Development%20of%20HTM_Manuscript-20250402.docx"
  }
]
                                        

                                            [
  "Methods development",
  "Sample Processing",
  "anthrax",
  "emergency response",
  "high throughput"
]
                                        

                                            [
  "020:000"
]
                                        

                                            {
  "name": "U.S. EPA Office of Research and Development (ORD)",
  "subOrganizationOf": {
    "name": "U.S. Environmental Protection Agency",
    "subOrganizationOf": {
      "name": "U.S. Government"
    }
  }
}
                                        

                                            [
  "https://doi.org/10.1016/j.mimet.2025.107149"
]
                                        

                                            null