Abstract
Pneumonic plague is a lethal infectious disease caused by Yersinia pestis, a Tier-1 biothreat agent. Antibiotic treatment can save infected patients; however, therapy should begin within 24 h of symptom onset. As some Y. pestis strains showed an antibiotic resistance phenotype, an antibiotic susceptibility test (AST) must be performed. Performing the Clinical and Laboratory Standards Institute (CLSI)-recommended standard process, which includes bacterial isolation, enumeration and microdilution testing, lasts several days. Thus, rapid AST must be developed. As previously published, the Y. pestis-specific reporter phage ϕA1122::luxAB can serve for rapid identification and AST (ID-AST). Herein, we demonstrate the ability to use ϕA1122::luxAB to determine minimal inhibitory concentration (MIC) values and antibiotic susceptibility categories for various Y. pestis therapeutic antibiotics. We confirmed the assay by testing several nonvirulent Y. pestis isolates with reduced susceptibility to doxycycline or ciprofloxacin. Moreover, the assay can be performed directly on positive human blood cultures. Furthermore, as Y. pestis may naturally or deliberately be spread in the environment, we demonstrate the compatibility of this direct method for this scenario. This direct phage-based ID-AST shortens the time needed for standard AST to less than a day, enabling rapid and correct treatment, which may also prevent the spread of the disease.
Highlights
Yersinia pestis, the causative agent of plague disease, is a highly lethal pathogen [1]that impacted the history of humankind through three worldwide pandemics [2,3]
A positive luminescent signal was defined as 100 relative luminescent unit/relative luminescent units (RLUs) (2.5 × background luminescent signal of growth media only)
We found that detection of Y. pestis directly from environmental samples using the bioluminescent reporter phage resulted in lower luminescent signal values (RLU)
Summary
The causative agent of plague disease, is a highly lethal pathogen [1]that impacted the history of humankind through three worldwide pandemics [2,3]. The causative agent of plague disease, is a highly lethal pathogen [1]. The last large outbreak of plague took place in Madagascar in. 2017, where many of the patients conducted pneumonic plague [4]. Pneumonic plague is a severe form of the disease, demonstrating 100% mortality in untreated patients [5]. It can cause death even in properly antibiotic-treated patients [6]. A recently report describing the death rate of probable or confirmed pneumonic plaque in the 2017 plague outbreak that occurred in Madagascar was 8–25% [7]
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