title: Application of Nuclear Magnetic Resonance to Detect Toxigenic Clostridium difficile from Stool Specimens: A Proof of Concept.
abstract: We evaluated the performance of an early prototype core molecular mirroring nuclear magnetic resonance detection platform (Mentor-100) to detect toxigenic Clostridium difficile from stool. This technology uses customized nanoparticles bound to target specific oligonucleotide probes that form binaries in the presence of nucleic acid from the target microorganism. Liquid patient stool specimens were seeded with C. difficile or other Clostridium species to determine the analytical sensitivity and specificity. Samples underwent nucleic acid extraction and target amplification with probes conjugated with iron nanoparticles. Signal from nuclear magnetic resonance spin-spin relaxation time was measured to detect the presence or absence of toxigenic C. difficile. The limit of detection was <180 colony forming units per reaction of toxigenic C. difficile. No cross-reactivity was observed with nontoxigenic C. difficile, Clostridium sordellii, Clostridium perfringens, Bacillus subtilis, or Paenibacillus polymyxa at 108colony forming units/mL. Correlation studies using frozen stool samples yielded a sensitivity of 88.4% (61 of 69) and a specificity of 87.0% (40 of 46) as compared with a commercial PCR assay for C. difficile. The area under the curve in the receiver operating characteristic curve analysis was 0.922. The prototype molecular mirroring platform showed promising performance for pathogen detection from clinical specimens. The platform design has the potential to offer a novel, low-cost alternative to currently available nucleic acid-based tests.
authors.affiliation: Menon Biosensors, Inc., San Diego, California.
authors.forename: Paul
authors.initials: P
authors.lastname: Yang
authors.affiliation: Menon Biosensors, Inc., San Diego, California.
authors.forename: Sara
authors.initials: S
authors.lastname: Hash
authors.affiliation: Keck School of Medicine, University of Southern California, Los Angeles, California.
authors.forename: Katherine
authors.initials: K
authors.lastname: Park
authors.affiliation: Menon Biosensors, Inc., San Diego, California.
authors.forename: Charlene
authors.initials: C
authors.lastname: Wong
authors.affiliation: Menon Biosensors, Inc., San Diego, California.
authors.forename: Loganathan
authors.initials: L
authors.lastname: Doraisamy
authors.affiliation: Keck School of Medicine, University of Southern California, Los Angeles, California.
authors.forename: Jonas
authors.initials: J
authors.lastname: Petterson
authors.affiliation: HealthSpring Global, Inc., Bradenton, Florida.
authors.forename: Cathy A
authors.initials: CA
authors.lastname: Petti
authors.affiliation: Keck School of Medicine, University of Southern California, Los Angeles, California.
authors.forename: Pamela M
authors.initials: PM
authors.lastname: Ward
authors.affiliation: Keck School of Medicine, University of Southern California, Los Angeles, California.
authors.forename: Seung Heon
authors.initials: SH
authors.lastname: Lee
authors.affiliation: Menon Biosensors, Inc., San Diego, California.
authors.forename: Suresh
authors.initials: S
authors.lastname: Menon
authors.affiliation: Keck School of Medicine, University of Southern California, Los Angeles, California. Electronic address: rosemary.she@med.usc.edu.
authors.forename: Rosemary C
authors.initials: RC
authors.lastname: She
mesh_terms: D001427:Bacterial Toxins / Q000235:genetics*; D016360:Clostridioides difficile / Q000235:genetics*; D003015:Clostridium Infections / Q000175:diagnosis* / Q000382:microbiology*; D005243:Feces / Q000382:microbiology*; D006801:Humans; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D053758:Nanoparticles; D036103:Nanotechnology; D016133:Polymerase Chain Reaction; D012372:ROC Curve; D012680:Sensitivity and Specificity
chemical_list: D001427:Bacterial Toxins