title: Demonstration of the Pre-Emergency Use Authorization Path Using 3 Minor Groove Binder-Hydrolysis Probe Assays to Detect Escherichia coli O104:H4.
abstract: BACKGROUND
  The Department of Defense (DoD) and the Food and Drug Administration (FDA) have collaboratively worked on a pre-Emergency Use Authorization (pre-EUA) process for in vitro diagnostic (IVD) devices, using FDA's regulatory flexibilities under the EUA authorities. The pre-EUA process enables FDA review of data in anticipation of a request for an EUA, advancing US government public health emergency preparedness efforts.
  
  
  METHODS
  The IVD device developed to detect Escherichia coli O104:H4, for which an EUA has not been issued, serves as an example to illustrate that process. Specifically, DoD designed real-time PCR assays to target the virulent E. coli strain O104:H4 (etiological agent of the 2011 German outbreak) including: fliC (flagellin), Agg3C (AAF), and rfb (wbwC) on the basis of the published sequences.
  
  
  RESULTS
  After development and optimization of these 3 specific assays, a defined protocol was followed to determine and document the sensitivity and specificity of each assay analytically.
  
  
  CONCLUSIONS
  FDA reviewed these data and returned commentary on additional required experiments to complete the pre-EUA process and expedite the use of the device should there be an emergency need for an IVD device to detect this virulent E. coli strain before such a test is cleared by FDA.
authors.affiliation: Diagnostic Systems Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD; Clinical RM, Hinckley, OH.
authors.forename: Laurie J
authors.initials: LJ
authors.lastname: Hartman
authors.affiliation: Diagnostic Systems Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD;
authors.forename: Megan L
authors.initials: ML
authors.lastname: Heinrich
authors.affiliation: Diagnostic Systems Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD;
authors.forename: Ashley M
authors.initials: AM
authors.lastname: Zovanyi
authors.affiliation: Diagnostic Systems Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD;
authors.forename: Michael F
authors.initials: MF
authors.lastname: Ingram
authors.affiliation: Food and Drug Administration, Silver Spring, MD.
authors.forename: J Peyton
authors.initials: JP
authors.lastname: Hobson
authors.affiliation: Diagnostic Systems Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD;
authors.forename: David A
authors.initials: DA
authors.lastname: Kulesh
authors.affiliation: Diagnostic Systems Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD;
authors.forename: Philip D
authors.initials: PD
authors.lastname: Craw
authors.affiliation: Diagnostic Systems Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD;
authors.forename: James G
authors.initials: JG
authors.lastname: Jaissle
authors.affiliation: Diagnostic Systems Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD;
authors.forename: David A
authors.initials: DA
authors.lastname: Norwood
authors.affiliation: Diagnostic Systems Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD; timothy.d.minogue.civ@mail.mil.
authors.forename: Timothy D
authors.initials: TD
authors.lastname: Minogue
mesh_terms: D004269:DNA, Bacterial / Q000235:genetics; D004196:Disease Outbreaks; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification*; D004927:Escherichia coli Infections / Q000175:diagnosis* / Q000382:microbiology*; D029968:Escherichia coli Proteins / Q000235:genetics; D036781:Fimbriae Proteins / Q000235:genetics; D005408:Flagellin / Q000235:genetics; D005700:Galactosyltransferases / Q000235:genetics; D006801:Humans; D006868:Hydrolysis; D057230:Limit of Detection; D060888:Real-Time Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods; D017422:Sequence Analysis, DNA; D014481:United States; D014486:United States Food and Drug Administration
chemical_list: C079725:AggA protein, E coli; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D005408:Flagellin; D036781:Fimbriae Proteins; D005700:Galactosyltransferases; C000592114:WbwC protein, E coli