title: The utility of cerebrospinal fluid for the molecular diagnosis of toxoplasmic encephalitis.
abstract: The aim of this study was to assess the efficacy of nested polymerase chain reaction (PCR) and the loop-mediated isothermal amplification (LAMP) assay, which were developed to detect and identify toxoplasma parasites in human cerebrospinal fluid (CSF). Nested PCR was performed using primers generated by Dr. L.D. Sibley to target the 18S rDNA instead of the conventionally used primers which target the B1 gene. We also designed Toxoplasma gondii-specific LAMP primers targeting both genes. In vitro detection sensitivity was evaluated using 10-fold serially diluted genomic DNA purified from RH tachyzoites, and clinical sensitivity and specificity were evaluated using clinical CSF samples from 16 patients with toxoplasmic encephalitis (TE) and from 12 patients with other diseases. The 18S rDNA nested PCR showed the highest detection sensitivity limit with a minimum of 1.0 × 10(-8) ng/μL. However, sensitivity and specificity of nested PCR with clinical specimens were 50% and 100%, respectively. The sensitivity of molecular diagnosis of TE is not sufficient; therefore, patients clinically suspected of having TE should be treated promptly. Our molecular diagnostic tool would restrictively facilitate a definitive diagnosis of TE at an early stage in approximately 50% of patients.
authors.affiliation: Department of Infectious Diseases and Pulmonary Medicine, National Defense Medical College, 3-2 Namiki, Tokorozawa City, Saitama, Japan.
authors.forename: Kei
authors.initials: K
authors.lastname: Mikita
authors.forename: Takuya
authors.initials: T
authors.lastname: Maeda
authors.forename: Takeshi
authors.initials: T
authors.lastname: Ono
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Miyahira
authors.forename: Takashi
authors.initials: T
authors.lastname: Asai
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kawana
mesh_terms: D016054:DNA, Protozoan / Q000134:cerebrospinal fluid* / Q000737:chemistry; D004660:Encephalitis / Q000134:cerebrospinal fluid* / Q000175:diagnosis / Q000469:parasitology; D017125:Genes, Protozoan / Q000235:genetics; D006801:Humans; D057230:Limit of Detection; D025202:Molecular Diagnostic Techniques / Q000379:methods; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction / Q000379:methods*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D014122:Toxoplasma / Q000235:genetics / Q000302:isolation & purification*; D016781:Toxoplasmosis, Cerebral / Q000134:cerebrospinal fluid* / Q000175:diagnosis / Q000469:parasitology
chemical_list: D016054:DNA, Protozoan; D012337:RNA, Ribosomal, 18S