title: A novel subtyping assay for detection of Clostridium difficile virulence genes.
abstract: This proof-of-concept study demonstrates the application of a novel nucleic acid detection platform to detect Clostridium difficile in subjects presenting with acute diarrheal symptoms. This method amplifies three genes associated with C. difficile infection, including genes and deletions (cdtB and tcdC) associated with hypervirulence attributed to the NAP1/027/BI strain. Amplification of DNA from the tcdB, tcdC, and cdtB genes was performed using a droplet-based sandwich platform with quantitative real-time PCR in microliter droplets to detect and identify the amplified fragments of DNA. The device and identification system are simple in design and can be integrated as a point-of-care test to help rapidly detect and identify C. difficile strains that pose significant health threats in hospitals and other health-care communities.
authors.affiliation: Center for Biomedical Engineering, School of Engineering, Brown University, Providence, Rhode Island.
authors.forename: Stephanie L
authors.initials: SL
authors.lastname: Angione
authors.affiliation: Center for Biomedical Engineering, School of Engineering, Brown University, Providence, Rhode Island.
authors.forename: Aartik A
authors.initials: AA
authors.lastname: Sarma
authors.affiliation: Warren Alpert Medical School, Brown University, Providence, Rhode Island.
authors.forename: Aleksey
authors.initials: A
authors.lastname: Novikov
authors.affiliation: Division of Biology and Medicine, Brown University, Providence, Rhode Island.
authors.forename: Leah
authors.initials: L
authors.lastname: Seward
authors.affiliation: Division of Biology and Medicine, Brown University, Providence, Rhode Island.
authors.forename: Jennifer H
authors.initials: JH
authors.lastname: Fieber
authors.affiliation: Center for Biomedical Engineering, School of Engineering, Brown University, Providence, Rhode Island; Division of Infectious Diseases, Rhode Island Hospital, Providence, Rhode Island.
authors.forename: Leonard A
authors.initials: LA
authors.lastname: Mermel
authors.affiliation: Center for Biomedical Engineering, School of Engineering, Brown University, Providence, Rhode Island. Electronic address: anubhav_tripathi@brown.edu.
authors.forename: Anubhav
authors.initials: A
authors.lastname: Tripathi
mesh_terms: D036002:ADP Ribose Transferases / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D001427:Bacterial Toxins / Q000235:genetics; D016360:Clostridioides difficile / Q000235:genetics*; D004761:Enterocolitis, Pseudomembranous / Q000175:diagnosis; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods; D019095:Point-of-Care Systems; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014774:Virulence / Q000235:genetics*; D037521:Virulence Factors / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; D037521:Virulence Factors; C031815:toxB protein, Clostridium difficile; D036002:ADP Ribose Transferases; C410164:actin-specific ADP-ribosyltransferase, Clostridium