Rapid Susceptibility Testing from Blood-Derived Samples
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Filbrun, Alexandra
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Abstract
Bacterial bloodstream infections contribute to global mortality and morbidity in large part due to the rapid development of antimicrobial resistance. Constrained to bacterial burdens of 1-10 colony-forming-units of bacteria per milliliter of blood (~109 blood cells), conventional diagnosis relies on lengthy culture-amplification and -isolation prior to identification and antibiotic susceptibility testing (ASTs). The resulting >60-hour time-to-result is at odds with the only indicator of survival – time to initiation of appropriate treatment. Consequently, new technologies capable of rapidly establishing actionable treatment from blood-derived samples is clinically important to promote positive patient outcomes and stewardship of antimicrobial resistance. To address this need, we developed a novel separation technology based on selective hemolysis coupled with centrifugation through inverted tubes containing sucrose. Demonstrated on the top BSI-causing bacterial families of Enterobacteriaceae (E. coli), non-fermenting gram-negative rods (P. aeruginosia), and Staphylococcus (S. aureus), near-pure bacteria is recovered from positive blood culture media in ≤15 minutes with minimal user intervention. Antibiotic susceptibilities are readily established using our label-free, flow cytometric approach for results in as little as 5 hours, post blood culture positivity. Overall, this direct from positive blood cultures enables results more than one day sooner than conventional methods.
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2021-07-28
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Dissertation