Journal of Medical Virology · 2003 · 170 citations · 28 references
Life SciencesInfectious DiseasesMolecular VirologyNeuroinfectious DiseasesVirologyAnimal Infectious DiseasesAnimal VirusesViral InfectionsZoonotic Disease DiagnosisClinical Signs DetectionDirect Fluorescent Antibody TestingIndirect Fluorescent Antibody AssayNested Reverse Transcription Polymerase Chain ReactionPositive Clinical SignsRabies VirusReverse Transcription Polymerase Chain Reaction (Rt-pcr)Viral Disease Diagnosis
A 55-year-old bat conservationist was admitted to Ninewells Hospital, Dundee, Scotland, on November 11, 2002, with an acute haematemesis. He gave a 5-day history of pain and paraesthesia in the left arm, followed by increasing weakness of his limbs with evidence of an evolving encephalitis with cerebellar involvement. The patient had never been vaccinated against rabies and did not receive postexposure treatment. Using a hemi-nested reverse transcriptase-polymerase chain reaction (RT-PCR), saliva samples taken intravitam from different dates proved positive for rabies. A 400-bp region of the nucleoprotein gene was sequenced for confirmation and identified a strain of European bat lyssavirus (EBLV) type 2a. The diagnosis was confirmed using the fluorescent antibody test (FAT) and by RT-PCR on three brain samples (cerebellum, medulla, and hippocampus) taken at autopsy. In addition, a mouse inoculation test (MIT) was performed. Between 13 and 17 days postinfection, clinical signs of a rabies-like illness had developed in all five inoculated mice. Brain smears from each infected animal were positive by the FAT and viable virus was isolated. This fatal incident is only the second confirmed case of an EBLV type-2 infection in a human after exposure to bats.
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Julie Thompson, Desmond G. Higgins, Toby J. Gibson · Nucleic Acids Research · 1994 · 64.9K citations · Full text
Avoiding false positives with PCR
Shirley Kwok, Russell Higuchi · Nature · 1989 · 3.5K citations