A duplex PCR assay for detection and genotyping of Herpes simplex virus in cerebrospinal fluid.
A duplex polymerase chain reaction (PCR) assay for the detection and genotyping of Herpes simplex virus (HSV) 1 and 2 from cerebrospinal fluid (CFS) of infants was developed. The glycoprotein D (gD) gene of HSV was selected as a target for amplification. The assay is highly specific, sensitive and reproducible. Herpes simplex virus detection is performed by agarose gel electrophoresis and Southern blot using a chemiluminescent probe. The probe hybridizes to sequences common to both HSV-1 and 2. A DNA fragment of HSV gD gene was cloned and used as positive control and to determine the specificity and sensitivity of the assay. The PCR assay is user-friendly and unambiguously differentiates in one-step both herpes virus strains. The assay is useful to screen CFS specimens from infants exposed to HSV during birth and at risk of developing encephalitis.
González-Villaseñor
L I
LI
BBI-Biotech Research Laboratories, Gaithersburg, MD, 20877, USA.
D017931:DNA Primers; D015342:DNA Probes; D005814:Genes, Viral; D005838:Genotype; D006558:Herpes Genitalis / Q000134:cerebrospinal fluid / Q000821:virology; D006561:Herpes Simplex / Q000134:cerebrospinal fluid / Q000821:virology; D018259:Herpesvirus 1, Human / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D018258:Herpesvirus 2, Human / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D007223:Infant; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014759:Viral Envelope Proteins / Q000235:genetics*
D017931:DNA Primers; D015342:DNA Probes; D014759:Viral Envelope Proteins; C039447:glycoprotein D, Human herpesvirus 1; C040570:glycoprotein D-herpes simplex virus type 2