title: Macroarray assay for differential diagnosis of meningoencephalitis in southern Africa.
abstract: BACKGROUND
  Many cases of acute febrile illness with nervous manifestations go undiagnosed, partly because the potential pathogens are not investigated routinely.
  
  
  OBJECTIVE
  To develop a multiplex PCR-based macroarray for detection of 29 pathogens associated with febrile disease, aseptic meningitis and meningoencephalitis in southern Africa, including common viruses, bacteria, parasites and selected arboviruses.
  
  
  METHODS
  Pathogens were identified by hybridization of PCR amplicons to probes on a macroarray chip, followed by colorimetric detection.
  
  
  RESULTS
  Positive control specimens for all 29 targets were detected with high sensitivity. Twenty-seven clinical samples previously found positive for various etiologies of febrile disease and meningoencephalitis, including less common infections such as Crimean Congo haemorrhagic fever, Rift Valley fever, West Nile and rabies were all identified. Testing of a blinded panel of 16 specimens in triplicate demonstrated high repeatability. Screening of 138 specimens from patients with febrile and/or neurological signs that could not be solved in routine investigations yielded 5 additional diagnoses.
  
  
  CONCLUSIONS
  A highly sensitive, specific and easily interpreted test was developed to detect 29 pathogens associated with febrile and nervous disease.
authors.affiliation: Zoonoses Research Unit, University of Pretoria, Pretoria, South Africa; Centre for Respiratory Diseases and Meningitis, National Institute for Communicable Diseases of the National Health Laboratory Service, Sandringham, South Africa. Electronic address: marietjiev@nicd.ac.za.
authors.forename: Marietjie
authors.initials: M
authors.lastname: Venter
authors.affiliation: Zoonoses Research Unit, University of Pretoria, Pretoria, South Africa.
authors.forename: Dewald
authors.initials: D
authors.lastname: Zaayman
authors.affiliation: Zoonoses Research Unit, University of Pretoria, Pretoria, South Africa.
authors.forename: Stephanie
authors.initials: S
authors.lastname: van Niekerk
authors.affiliation: Zoonoses Research Unit, University of Pretoria, Pretoria, South Africa.
authors.forename: Voula
authors.initials: V
authors.lastname: Stivaktas
authors.affiliation: Zoonoses Research Unit, University of Pretoria, Pretoria, South Africa.
authors.forename: Shivani
authors.initials: S
authors.lastname: Goolab
authors.affiliation: Special Viral Pathogens Reference Laboratory, Centre for Emerging and Zoonotic Diseases, National Institute for Communicable Diseases of the National Health Laboratory Service, Sandringham, South Africa.
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Weyer
authors.affiliation: Special Viral Pathogens Reference Laboratory, Centre for Emerging and Zoonotic Diseases, National Institute for Communicable Diseases of the National Health Laboratory Service, Sandringham, South Africa.
authors.forename: Janusz T
authors.initials: JT
authors.lastname: Paweska
authors.affiliation: Zoonoses Research Unit, University of Pretoria, Pretoria, South Africa.
authors.forename: Robert
authors.initials: R
authors.lastname: Swanepoel
mesh_terms: D000353:Africa, Southern; D000818:Animals; D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D003124:Colorimetry / Q000379:methods; D003937:Diagnosis, Differential; D006801:Humans; D008590:Meningoencephalitis / Q000175:diagnosis* / Q000209:etiology*; D046228:Microarray Analysis / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009693:Nucleic Acid Hybridization / Q000379:methods; D010271:Parasites / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014780:Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification
keywords: Differential diagnosis; Macroarray; Meningoencephalitis multiplex PCR