title: Microsphere suspension array assays for detection and differentiation of Hendra and Nipah viruses.
abstract: Microsphere suspension array systems enable the simultaneous fluorescent identification of multiple separate nucleotide targets in a single reaction. We have utilized commercially available oligo-tagged microspheres (Luminex MagPlex-TAG) to construct and evaluate multiplexed assays for the detection and differentiation of Hendra virus (HeV) and Nipah virus (NiV). Both these agents are bat-borne zoonotic paramyxoviruses of increasing concern for veterinary and human health. Assays were developed targeting multiple sites within the nucleoprotein (N) and phosphoprotein (P) encoding genes. The relative specificities and sensitivities of the assays were determined using reference isolates of each virus type, samples from experimentally infected horses, and archival veterinary diagnostic submissions. Results were assessed in direct comparison with an established qPCR. The microsphere array assays achieved unequivocal differentiation of HeV and NiV and the sensitivity of HeV detection was comparable to qPCR, indicating high analytical and diagnostic specificity and sensitivity.
authors.affiliation: Australian Animal Health Laboratory, CSIRO Animal, Food and Health Sciences, Geelong, VIC 3220, Australia.
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Foord
authors.forename: John R
authors.initials: JR
authors.lastname: White
authors.forename: Axel
authors.initials: A
authors.lastname: Colling
authors.forename: Hans G
authors.initials: HG
authors.lastname: Heine
mesh_terms: D000818:Animals; D004279:DNA, Viral / Q000032:analysis; D004196:Disease Outbreaks; D045404:Hendra Virus / Q000302:isolation & purification*; D045464:Henipavirus Infections / Q000175:diagnosis* / Q000662:veterinary* / Q000821:virology; D006736:Horses; D008863:Microspheres; D045405:Nipah Virus / Q000302:isolation & purification*; D009698:Nucleoproteins / Q000032:analysis; D010750:Phosphoproteins / Q000032:analysis; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis; D012372:ROC Curve; D014759:Viral Envelope Proteins / Q000032:analysis
chemical_list: D004279:DNA, Viral; D009698:Nucleoproteins; D010750:Phosphoproteins; D012367:RNA, Viral; D014759:Viral Envelope Proteins