Diagnosis of herpes encephalitis via Southern blotting of cerebrospinal fluid DNA amplified by polymerase chain reaction

Abstract

Herpes simplex virus thymidine kinase gene specific polymerase chain reaction (PCR) amplification of DNA extracted from lumbar cerebrospinal fluid (CSF) and Southern blotting (SB) were evaluated as a method for the diagnosis of herpes simplex encephalitis (HSE). Positive PCR-SB results were obtained with CSF samples from 9 of 10 patients (11 of 12 CSF specimens) with proven herpes encephalitis as early as 2 days after onset of neurological illness. Our data support the suggestion that PCR techniques may provide a clinically relevant "non-invasive" method for the diagnosis of HSE.

Herpes Simplex Virus Meningitis Complicated by Ascending Paralysis

“…Th e CSF profi le typically includes a lymphocytic pleocytosis, a normal CSF glucose concentration, and elevated protein. HSV DNA detection by polymerase chain reaction in the serum and/ or CSF is the most sensitive and specifi c method for detecting HSV meningitis (8).…”

Herpes Simplex Encephalitis Presenting with Bilateral Hippocampal Lesions on Magnetic Resonance Imaging, Simultaneously Complicated by Small Cell Lung Carcinoma

“…ers were used at both the DNA polymerase and thymidine kinase gene regions of HSV type 1 (9). HSV type 1 DNA was detected in CSF at both gene regions.…”

Pulmonary herpes simplex in burns patients

“…Precipitated DNA was resuspended in 50 µL of sterile distilled water and 5 µL of this was used in the PCR. The PCR reaction mixture consisted of 100 nM of each oligonucleotide primer (oligonucleotide primers which amplify a 351 bp fragment within the HSV-1 TK gene [12]), 200 µmol deoxynucleotide triphosphates (dNTPs), 0.05 M KCl, 2 mM MgCl 2, , 0.02% bovine serum albumin (BSA) and 2.5 units of Taq polymerase in 0.01 M Tris-HCl at pH 8.3. The reaction mixtures were subjected to 50 cycles at 94°C for 2 min, 50°C for 1.5 min, and 70°C for 2 min.…”

“…The reaction mixtures were subjected to 50 cycles at 94°C for 2 min, 50°C for 1.5 min, and 70°C for 2 min. Reaction products were analysed by agarose gel electrophoresis and Southern blotting, using a 128 bp probe for a region within the 351 bp amplicon [12].…”