title: Real-time Leishmania genus master mix: a platform compatibility and stability study.
abstract: Performing diagnostics and vector-pathogen surveillance in austere environments is challenging. On-site diagnostic/detection mitigates vector-borne disease complications during military or humanitarian deployments to disease endemic locals. The mobile molecular diagnostic platform, Joint Biological Agent Identification and Diagnostic System (JBAIDS; BioFire Diagnostics Inc., Salt Lake City, UT, USA), rapidly identifies biothreat pathogens. Although ideal for remote diagnostics, the platform was validated for specific pathogens of insignificant epidemiological consequence. Recognizing the JBAIDS's remote diagnostic/detection versatility, we tested a Leishmania genus real-time PCR master mix validated for use on the SmartCycler® (Cepheid, Sunnyvale, CA, USA) for concomitant use on the JBAIDS. We evaluated assay sensitivity, precision, and specificity of one or more Leishmania spp. on the JBAIDS and found that the JBAIDS produces superior detection sensitivity and specificity compared to the SmartCycler®. We also examined the storage stability of a bulk lot preparation of the Leishmania genus real-time PCR master mix on the SmartCycler® to ensure that long periods of frozen storage that would translate to a field environment with the JBAIDS were not detrimental to the reagent. We found that the bulk master mix maintains its stability over a 13-month time period. Overall, these studies confirm JBAIDS's versatility and demonstrate a streamlined assay development approach where reagents are compatible with both platforms.
authors.affiliation: Diagnostics and Laboratory Services Department, Entomology Branch, Walter Reed Army Institute of Research, 503 Robert Grant Ave, Silver Spring, MD 20910. Electronic address: vanessa.melanson@us.army.mil.
authors.forename: Vanessa R
authors.initials: VR
authors.lastname: Melanson
authors.affiliation: Diagnostics and Laboratory Services Department, Entomology Branch, Walter Reed Army Institute of Research, 503 Robert Grant Ave, Silver Spring, MD 20910. Electronic address: jessica.l.scheirer.ctr@mail.mil.
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Scheirer
authors.affiliation: Diagnostic Systems Division, Systems Development Department, United States Army Research Institute of Infectious Disease, 1425 Porter St, Fort Detrick, MD 21702. Electronic address: warren.kalina@us.army.mil.
authors.forename: Warren V
authors.initials: WV
authors.lastname: Kalina
authors.affiliation: Wound Infections Department, Bacterial Diseases Branch, Walter Reed Army Institute of Research, 503 Robert Grant Ave, Silver Spring, MD 20910. Electronic address: eric.j.wagar2.mil@mail.mil.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Wagar
mesh_terms: D006801:Humans; D007891:Leishmania / Q000235:genetics / Q000302:isolation & purification*; D007896:Leishmaniasis / Q000175:diagnosis* / Q000469:parasitology; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D010274:Parasitology / Q000379:methods*; D019095:Point-of-Care Systems; D011933:Reagent Kits, Diagnostic; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D011933:Reagent Kits, Diagnostic
keywords: Leishmania; Master mix; Platform comparison; Real-time PCR; Stability