A Prospective Study of the Polymerase Chain Reaction for Detection of Herpes Simplex Virus in Cerebrospinal Fluid Submitted to the Clinical Virology Laboratory

Mol Cell Probes. 1992 Oct;6(5):367-73. doi: 10.1016/0890-8508(92)90029-w.

Abstract

Polymerase chain reaction (PCR) was prospectively performed with cerebrospinal fluid (CSF) from 51 patients whose CSF was available for analysis and was submitted for viral culture and/or herpes simplex virus (HSV) serology and 20 patients whose CSF was submitted exclusively to the Clinical Biochemistry Laboratory. Primers were used that flanked a 92 bp segment of the HSV DNA polymerase gene (35 cycles). Amplified products were electrophoresed on agarose gel, blotted onto nylon membrane, and probed with a 32P-labelled sequence internal to the primers. For nested PCR, 1 microliter of PCR product was amplified for an additional 35 cycles before electrophoresis and Southern blot analysis. Review of the clinical records revealed that 15 patients had central nervous system (CNS) infections. Specific HSV DNA sequences were detected in CSF specimens of three of the individuals [PCR(2), nested PCR(1)]. Two of these patients had disseminated HSV infection including encephalitis and one patient had aseptic meningitis. The diagnoses of the 12 patients with CNS infection who did not have HSV DNA detected in CSF included encephalitis [varicella-zoster virus (1), cytomegalovirus (1), Mycoplasma pneumoniae (1)], meningitis [Neisseria meningitidis (1), Coccidioides immitis (1), Enterovirus (1), aseptic meningitis (1)], varicella-zoster radiculitis (2), human immunodeficiency virus dementia (2), and transverse myelitis due to Epstein-Barr virus (1). Importantly, HSV DNA was also not detected in the CSF of the 36 patients who did not have CNS infection and 20 samples submitted exclusively to the Clinical Biochemistry Laboratory. Our findings demonstrate the utility of PCR as a rapid, non-invasive method for the routine laboratory diagnosis of CNS infection due to HSV.

MeSH terms

  • Adult
  • Aged
  • Base Sequence
  • Cerebrospinal Fluid / microbiology
  • DNA-Directed DNA Polymerase / genetics
  • Encephalitis / cerebrospinal fluid
  • Encephalitis / epidemiology
  • Encephalitis / microbiology*
  • Exodeoxyribonucleases / genetics
  • Female
  • Herpes Simplex / cerebrospinal fluid
  • Herpes Simplex / epidemiology
  • Herpes Simplex / microbiology*
  • Humans
  • Male
  • Molecular Sequence Data
  • Oligodeoxyribonucleotides
  • Oligonucleotide Probes
  • Polymerase Chain Reaction* / methods
  • Pregnancy
  • Pregnancy Complications, Infectious / cerebrospinal fluid
  • Pregnancy Complications, Infectious / microbiology
  • Prospective Studies
  • Sensitivity and Specificity
  • Simplexvirus / genetics
  • Simplexvirus / isolation & purification*
  • Templates, Genetic
  • Viral Proteins*

Substances

  • Oligodeoxyribonucleotides
  • Oligonucleotide Probes
  • Viral Proteins
  • DNA-Directed DNA Polymerase
  • Exodeoxyribonucleases
  • DNA polymerase, Simplexvirus