Browsing by Author "Goux, Heather J."
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Item Aptamer-Phage Reporters for Ultrasensitive Lateral Flow Assays(Analytical Chemistry, 2016-12) Adhikari, Meena; Strych, Ulrich; Kim, Jinsu; Goux, Heather J.; Dhamane, Sagar; Poongavanam, Mohan-Vivekanandan; Hagstrom, Anna E. V.; Kourentzi, Katerina D.; Conrad, Jacinta C.; Willson, Richard C.We introduce the modification of bacteriophage particles with aptamers for use as bioanalytical reporters, and demonstrate the use of these particles in ultrasensitive lateral flow assays. M13 phage displaying an in vivo biotinylatable peptide (AviTag) genetically fused to the phage tail protein pIII were used as reporter particle scaffolds, with biotinylated aptamers attached via avidin朾iotin linkages, and horseradish peroxidase (HRP) reporter enzymes covalently attached to the pVIII coat protein. These modified viral nanoparticles were used in immunochromatographic sandwich assays for the direct detection of IgE and of the penicillin-binding protein from Staphylococcus aureus (PBP2a). We also developed an additional lateral flow assay for IgE, in which the analyte is sandwiched between immobilized anti-IgE antibodies and aptamer-bearing reporter phage modified with HRP. The limit of detection of this LFA was 0.13 ng/mL IgE, ?100 times lower than those of previously reported IgE assays.Item Development of a panel of recombinase polymerase amplification assays for detection of common bacterial urinary tract infection pathogens(Journal of Applied Microbiology, 8/1/2018) Raja, Balakrishnan; Goux, Heather J.; Marapadaga, Archana; Rajagopalan, Sri; Kourentzi, Katerina D.; Willson, Richard C.Aims: To develop and evaluate the performance of a panel of isothermal real?time recombinase polymerase amplification (RPA) assays for detection of common bacterial urinary tract infection (UTI) pathogens. Methods and Results: The panel included RPAs for Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa and Enterococcus faecalis. All five RPAs required reaction times of under 12 min to reach their lower limit of detection of 100 genomes per reaction or less, and did not cross?react with high concentrations of nontarget bacterial genomic DNA. In a 50?sample retrospective clinical study, the five?RPA assay panel was found to have a specificity of 100% (95% CI, 78�0%) and a sensitivity of 89% (95% CI, 75�%) for UTI detection. Conclusions:The analytical and clinical validity of RPA for the rapid and sensitive detection of common UTI pathogens was established. Significance and Impact of the Study: Rapid identification of the causative pathogens of UTIs can be valuable in preventing serious complications by helping avoid the empirical treatment necessitated by traditional urine culture's 48�?h turnaround time. The routine and widespread use of RPA to supplement or replace culture?based methods could profoundly impact UTI management and the emergence of multidrug?resistant pathogens.