Application of Competitive PCR to Cerebrospinal Fluid Samples from Patients with Herpes Simplex Encephalitis

Abstract

The purpose of the present study was to determine if the quantity of herpes simplex virus (HSV) DNA in the cerebrospinal fluid (CSF) of patients with herpes encephalitis would be useful in establishing the prognosis of the disease and to determine the effect of antiviral therapy on the clearance of viral DNA from the CSF. Quantitation of HSV DNA was done by constructing an internal standard (IS) from the glycoprotein B amplicon which had a 25-bp deletion between primer annealing sites. Each CSF specimen was coamplified with the IS and the ratio of the amount of HSV/amount of IS was compared to the ratios on a standard curve constructed with the same IS plus known amounts of HSV DNA. CSF specimens were available from 16 patients who were treated with intravenous acyclovir, and the amount of HSV DNA ranged from <25 to 18,000 copies per μl in CSF obtained before or within 4 days of the initiation of acyclovir therapy. Patients with >100 copies of HSV DNA per μl were older, were found by computed tomography to have lesions, and had poorer outcomes than patients with <100 copies. Follow-up CSF specimens were available from seven patients. In six of these seven patients, the HSV DNA levels decreased during therapy. One patient had a twofold increase in HSV DNA levels after 1 week of therapy and died on day 8. The application of this assay may be helpful in establishing the prognosis and in the monitoring of patients with herpes simplex encephalitis.

High variability in viral load in cerebrospinal fluid from patients with herpes simplex and varicella-zoster infections of the central nervous system

“…Domingues et al. [7] examined the potential utility of quantifying HSV DNA in CSF from HSE patients as a possible prognostic marker [7], and Kimura et al. [8] considered that quantification of the viral load in serum is useful for assessing the prognosis of neonatal HSV infections.…”

“…Dominigues et al. [7] found that patients with >100 copies of HSV DNA/μL of CSF had a less successful outcome than those with <100 copies, suggesting the possible value of quantitative measurement of HSV copies in CSF for disease prognosis. In contrast, Lakeman et al.…”

Diagnosis and surveillance of herpes simplex virus infection of the central nervous system

“…Recently, Domingues et al [1998b] applied a quantitative PCR assay to CSF samples collected from HSV encephalitis cases to determine if this technique has a potential use for monitoring of HSV encephalitis treatment. They analysed 16 cases, showing that in six cases with favourable outcome, they could monitor either a decrease in HSV burden in CSF (4 cases) or no HSV at all (2 cases), whereas 1 fatal case showed an increase in viral load in CSF before death.…”

Clinical Applications of Quantitative Real-Time PCR in Virology

“…There exists a clear application for quantitative real-time PCR technology in baseline determination of patients with significant viral infections, and in fact, quantitative viral load determination plays a critical role in therapeutic decision for many other viral infections. High baseline viral load has been shown to correlate with advanced disease during infection with numerous viruses such as BKV, HSV-1, EBV, and Adenovirus and may potentiate the need for longer duration therapies in certain scenarios (Cincinnati Children's Hospital Medical Center, 2012;Domingues, Lakeman, Mayo, & Whitley, 1998;Gustafson et al, 2008;Randhawa et al, 2004).…”