Scandinavian Journal of Infectious Diseases · 1993 · 28 citations · 24 references
Life SciencesInfectious DiseasesMicrobiologyDiagnostic MicrobiologyMedical MicrobiologyVirologyCentral Nervous System InfectionsCerebrovascular DiseaseInfectious Disease Molecular TestingNeurological Parasitic DiseasesCerebrospinal Fluid DiagnosisHigh-sensitivity AssaysNeurological Disease CasesToxoplasma DetectionToxoplasma Gondii Detection
Reactivation of Toxoplasma gondii can lead to a life-threatening intracerebral infection in immunocompromised HIV-positive patients. Due to the current diagnostic limitations for establishing an exact diagnosis of cerebral toxoplasmosis, a nested PCR system was developed for direct detection of T. gondii in cerebrospinal fluid. A storage temperature for samples of -20 degrees C and sample preparation using Proteinase K appeared to be critical for obtaining a high sensitivity of PCR. A total of 56 samples from 38 HIV-positive patients and 12 HIV-negative patients with symptoms or signs of neurological disease were evaluated by PCR. 5 of the 38 HIV-positive patients were diagnosed as having cerebral toxoplasmosis and PCR was positive in samples from all 5 patients. In the remaining 33, PCR was positive in one case and negative in 32. An exact etiological diagnosis other than cerebral toxoplasmosis was established in 5 patients. PCR performed on cerebrospinal fluid samples seems to be a fast, sensitive, specific and valuable tool for establishing the diagnosis of cerebral toxoplasmosis among HIV-positive patients at the time of presentation of symptoms or signs of neurological disease.
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Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA Polymerase
Randall K. Saiki, David H. Gelfand, Susanne Stoffel et al. · Science · 1988 · 17.2K citations
Avoiding false positives with PCR
Shirley Kwok, Russell Higuchi · Nature · 1989 · 3.5K citations