Abstract
Certain serovars of Salmonella enterica subsp. enterica cause invasive disease (e.g., enteric fever, bacteremia, septicemia, meningitis, etc.) in humans and constitute a global public health problem. A rapid, sensitive diagnostic test is needed to allow prompt initiation of therapy in individual patients and for measuring disease burden at the population level. An innovative and promising new rapid diagnostic technique is microwave-accelerated metal-enhanced fluorescence (MAMEF). We have adapted this assay platform to detect the chromosomal oriC locus common to all Salmonella enterica subsp. enterica serovars. We have shown efficient lysis of biologically relevant concentrations of Salmonella spp. suspended in bacteriological media using microwave-induced lysis. Following lysis and DNA release, as little as 1 CFU of Salmonella in 1 ml of medium can be detected in <30 seconds. Furthermore the assay is sensitive and specific: it can detect oriC from Salmonella serovars Typhi, Paratyphi A, Paratyphi B, Paratyphi C, Typhimurium, Enteritidis and Choleraesuis but does not detect Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Streptococcus pneumoniae, Haemophilus influenzae or Acinetobacter baumanii. We have also performed preliminary experiments using a synthetic Salmonella oriC oligonucleotide suspended in whole human blood and observed rapid detection when the sample was diluted 1∶1 with PBS. These pre-clinical data encourage progress to the next step to detect Salmonella in blood (and other ordinarily sterile, clinically relevant body fluids).
Highlights
Salmonella, a genus of more than 2500 serological variants, includes many organisms that can cause human disease
We describe adaption of the ultra-fast, highly sensitive microwave-accelerated metal-enhanced fluorescence (MAMEF) technology to detect the oriC locus, that we currently use as a Salmonella-specific target in polymerase chain reaction (PCR) [11], of Salmonella spp. which have been rapidly lysed by focused microwave radiation
Salmonella spp., E. coli, P. aeruginosa and K. pneumoniae were grown in an animal product-free (APF) LB Lennox medium (APF-LB; Athena Environmental Sciences, Baltimore, MD, USA) at 37uC
Summary
Salmonella, a genus of more than 2500 serological variants (serovars), includes many organisms that can cause human disease. Non-typhoidal Salmonella (NTS) generally produce a self-limited gastroenteritis (vomiting, fever and diarrhea) in healthy humans [2,3,4]. In young infants, the elderly and immunocompromised hosts, NTS can cause severe, fatal disease in both industrialized [4,5] and developing countries [2,6,7,8,9,10,11,12]. Incidence rates of 200–350 cases of invasive NTS infections/105 infections in infants and toddlers have been recorded with case fatality rates of 20–30% [10,13,14,16]
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