Year: 2016
Type: article
Abstract: There is currently no widely available optimal assay for diagnosing patients with enteric fever. Here we present a novel assay designed to detect amplified Salmonella nucleic acid (mRNA) using magneto-DNA probes and a miniaturized nuclear magnetic resonance device. We designed primers for genes specific to S. Typhi, S. Paratyphi A, and genes conserved among Salmonella enterica spp. and utilized strongly magnetized nanoparticles to enhance the detection signal. Blood samples spiked with in vitro grown S. Typhi, S. Paratyphi A, S. Typhimurium, and E. coli were used to confirm the specificity of each probe-set, and serial 10-fold dilutions were used to determine the limit of the detection of the assay, 0.01-1.0 CFU/ml. For proof of principle, we applied our assay to 0.5 mL blood samples from 5 patients with culture-confirmed enteric fever from Bangladesh in comparison to 3 healthy controls. We were able to detect amplified target cDNA in all 5 cases of enteric fever; no detectable signal was seen in the healthy controls. Our results suggest that a magneto-DNA nanoparticle system, with an assay time from blood collection of 3.5 hours, may be a promising platform for the rapid and culture-free diagnosis of enteric fever and non-typhoidal Salmonella bacteremia.
Source: Scientific Reports
DOI: 10.1038/srep32878
Language: English
Subfield: Biomedical Engineering
Field: Engineering
Domain: Physical Sciences
Open Access status: gold
Estimated APC paid: $2,690
Funders: Robert Wood Johnson Foundation, Styrelsen för Internationellt Utvecklingssamarbete, Massachusetts General Hospital, National Research Foundation, National Research Foundation of Korea, International Centre for Diarrhoeal Disease Research, Bangladesh, National Institutes of Health, National Heart, Lung, and Blood Institute, National Cancer Institute, National Institute of Allergy and Infectious Diseases, Fogarty International Center, National Institute of Biomedical Imaging and Bioengineering
Awards: Improving diagnostic capabilities for typhoid fever, G1098359285, Multiplexed analysis of exosomes in cancer nano therapy, G1510978416, Novel Clickdyes for Biomedical Sensing, G172571585, G2066243351, Analysis of scant cancer cells in fine needle aspirates, G435013092, High-throughput proteomic study of protective immunity in human cholera infection, ICDDRB research 2012-2015, G531883626, G5850682332, G6141571532, MAGNETIC NANOSENSORS FOR BIOMEDICAL ANALYSES OF MICROVESICLES, G7521024345, Chip NMR biosensor for molecular analysis of cells, International research training program in vaccine development and public health
OpenAlex ID: W2513669844