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In 1998 the US Commission on National Security in the 21st Century stated biological agents were the most likely weapon of choice for disaffected states and groups. The 2001 Anthrax letters case highlighted this threat of bacterial biological warfare agents, resulting in 5 deaths and a clean up costing hundreds of millions of US government dollars. Biological warfare agents (BWAs) appeal to disaffected states and groups due their ease of relatively cheap manufacture, small doses required for maximum damage, and difficulty to trace.
Being able to rapidly identify which bacterium was used in an incident would lead to faster, specific support to victims, as well as providing vital information to aid front line personnel. Within a healthcare environment knowing the specific identity of bacterium allows the correct antibiotics to be prescribed and gives important information on any necessary quarantine conditions required to prevent the spread of infection.
Forensically, strain level identification of BWAs can help determine the original laboratory source of bacteria and medically aid precise diagnosis and treatment. Strain level identification is currently very challenging and time consuming. Methods such as MALDI-TOF struggle with strain level identification due to the samples being too similar. Vibrational spectroscopies fingerprint of information and handheld instruments provides an exciting, novel opportunity to provide more detailed bacterial identification.
Bacillus anthracis presents itself as an effective bioweapon not only due to its mortality rates, but its ability to form spores. Spores protect the bacteria from harsher conditions and can remain dormant for vast lengths of time until effectively awoken when more desirable growth conditions. Gruinard Island is an examples of bacilli spores’ survival strength.
ATR-FTIR is a well proven technique for investigating biological molecules. However, one significant issue with the technique is the use of expensive fixed internal reflective elements (IRE’s), e.g. diamond, and thus drying times. Water is a strong infra-red absorber, resulting in the need to dry samples to reveal the detailed information lying underneath H2O peaks. Here, ATR-FTIR has been developed to encompass a more high throughput approach, ditching expensive IREs for cheap, disposable silicon IREs SIREs). With these IREs samples can be batch dried and analysed much more rapidly. Their disposable nature removes the need to clean in-between samples, reducing the risk of contamination. This work aims to differentiate bacilli samples capable of forming spores, both in their spore and vegetative state using this novel SIRE approach.
With the current turbulence seen amongst defect states and militant groups the threat of biowarfare being used is increasing and accurate, rapid identification is crucial. This paper reports the use of a novel method of high throughput vibrational spectroscopy to demonstrate the use of FTIR for analysis of bacteria. This work will present a bacteria study, involving biological warfare surrogates, with the aim of advancing FTIR into a higher throughput, rapid approach to bacterial identification which can identify down to the strain level.
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