Real-Time Polymerase Chain Reaction Detection of Herpes Simplex Virus in Cerebrospinal Fluid and Cost Savings from Earlier Hospital Discharge

Abstract

Neonatal herpes simplex virus (HSV) can be a devastating illness and may be difficult to diagnose in those cases without a typical skin rash. As a result, physicians often rely on HSV polymerase chain reaction of cerebrospinal fluid to rule out HSV encephalitis. We developed a real-time polymerase chain reaction assay for HSV using the SmartCycler II (Cepheid, Sunnyvale, CA). End point dilution studies showed sensitivity comparable to that of two national reference laboratories that use LightCycler. In-house turnaround time was Ϸ1.5 days versus Ϸ5.2 days for sending the test to a reference laboratory. We hypothesized that the rapid availability of a negative test result would allow physicians to discharge appropriate patients earlier. Six months after implementation, clinical case analysis identified 12 pediatric patients who were discharged earlier based on more rapid test results, with a projected savings of Ϸ55.2 hospital days throughout the first year. Actual length of stay for patients tested in-house was significantly less than that of historical controls and was projected to save Ϸ70.2 hospital days in the first year. Including projected annual laboratory cost/test savings of Ϸ$11,000, a total savings of $38,000 to $43,000 was estimated for the first year of implementation, more than offsetting startup instrument and development cost. (J Mol Diagn 2005, 7:511-516) Neonatal herpes simplex virus (HSV) infection is a life threatening infection occurring within the first several weeks after birth. Classically, ϳ60 to 70% of patients have a typical herpetic skin rash, with or without disseminated infection involving brain, liver, lung, and other organs.1 Because central nervous system HSV can present with seizures, low-grade fever, and other nonspecific symptoms in the absence of the vesicular skin rash, physicians often rely on a negative cerebrospinal fluid (CSF) HSV polymerase chain reaction (PCR) to rule out HSV encephalitis.2-4 In our hospital HSV CSF PCR was sent to a reference laboratory with an average 5.2-day turnaround time. We attempted to show that performing HSV CSF PCR in the hospital laboratory would shorten this turnaround time and lead to an earlier discharge for a subset of neonates without other complications who were kept in the hospital for observation pending this test result.

Viral encephalitis: a clinician’s guide

“…CSF viral culture is now rarely performed, although it has the potential advantage over PCR of being able to detect any virus, whereas PCR only detects the viruses being targeted by the panel of PCR assays used. Newer more sensitive PCR methods, such as ''real-time'' and quantitative PCR have improved the clinical utility of this test, and are becoming available for herpes viruses and enteroviruses 38. …”

Concepts for Lean Laboratory Organization

“…These needs influence the work in the lab by defining parameter spectrum, expected turnaround time, and sample size. On the other hand the lab influences the work in the clinical departments by influencing the time since diagnosis and therapy, guidance of therapy and length of hospital stay (16)(17)(18). Second, you have to measure the initial situation.…”

Simultaneous cytological diagnosis of herpes simplex virus infection and primary lung cancer: Report of two cases

“…Cycling conditions were an initial denaturation step at 958C for 5 minutes, amplification steps of 958C for 30 seconds, and 608C for 1 minutes for a total of 45 cycles. 10 From the examination of the amplification curves, we obtained a true positive result for the detection of HSV1. HSV2 was not detected.…”