title: Ebola Preparedness: Diagnosis Improvement Using Rapid Approaches for Proficiency Testing.
abstract: The unprecedented 2015 Ebolavirus (EBOV) outbreak in West Africa was declared a public health emergency, making diagnosis and quality of testing a global issue. The accuracy of laboratory diagnostic capacity for EBOV was assessed in 2014 to 2016 using a proficiency testing (PT) strategy developed by the Royal College of Pathologists of Australasia Quality Assurance Programs (RCPAQAP) in Biosecurity. Following a literature search, EBOV-specific gene targets were ranked according to the frequency of their use in published methods. The most commonly used gene regions (nucleoprotein [NP], glycoprotein [GP], and RNA-dependent RNA polymerase [L]) were selected for the design ofin vitroRNA transcripts to be included in the simulated EBOV specimens used for EBOV detection with PCR-based assays. Specimens were tested for stability and found to be stable on long-term storage (1 year) at -80°C and on shorter-term storage in lyophilized form (1 week at ambient temperature and a subsequent week at -80°C). These specimens were used in three EBOV PTs offered from April 2014 to March 2016. In the first and third PTs, all laboratories (3/3 and 9/9, respectively) correctly identified specimens containing EBOV RNA transcripts, while in the second PT, all but one laboratory (5/6) correctly confirmed the presence of EBOV. The EBOV PT panel was useful for ensuring the competency of laboratories in detecting EBOV in the absence of readily available clinical samples. The simulated EBOV specimen was safe, stable, and reliable and can be used in lyophilized form for future EBOV PT programs, allowing simplicity of transport.
authors.affiliation: Royal College of Pathologists of Australasia Quality Assurance Programs in Biosecurity, St. Leonards, NSW, Australia.
authors.forename: Katherine A
authors.initials: KA
authors.lastname: Lau
authors.affiliation: Royal College of Pathologists of Australasia Quality Assurance Programs in Biosecurity, St. Leonards, NSW, Australia.
authors.forename: Torsten
authors.initials: T
authors.lastname: Theis
authors.affiliation: Royal College of Pathologists of Australasia Quality Assurance Programs in Biosecurity, St. Leonards, NSW, Australia.
authors.forename: Joanna
authors.initials: J
authors.lastname: Gray
authors.affiliation: Serology and Virology Division, South Eastern Area Laboratory Services Microbiology, Prince of Wales Hospital, NSW Health Pathology, Sydney, NSW, Australia w.rawlinson@unsw.edu.au.|School of Medical Sciences and School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia.
authors.forename: William D
authors.initials: WD
authors.lastname: Rawlinson
mesh_terms: D044362:Australasia; D029043:Ebolavirus / Q000302:isolation & purification*; D019142:Hemorrhagic Fever, Ebola / Q000175:diagnosis*; D006801:Humans; D059021:Laboratory Proficiency Testing / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*
keywords: Ebola diagnosis; Ebolavirus; proficiency testing