Development of a reverse transcription‐polymerase chain reaction assay for diagnosis of lymphocytic choriomeningitis virus infection and its use in a prospective surveillance study

Abstract

Lymphocytic choriomeningitis virus (LCMV), which is one of several arenaviruses that are pathogenic for humans, causes encephalitis and meningitis in man. In this study, single-stage and nested reverse transcription-polymerase chain reaction (RT-PCR) assays were developed that targeted the GPC and N genes of LCMV. Both assays detected < 1 TCID50 unit of LCMV. These assays were used to measure the incidence of LCMV infection by testing cerebrospinal fluid (CSF) samples with > or = 10 leukocytes/microl collected over 1 year from patients undergoing lumbar puncture for diagnostic reasons at two Birmingham hospitals. Samples were tested for the presence of LCMV RNA by using the RT- PCR assay and for LCMV-specific IgM antibody by using an ELISA assay. None of the specimens collected from 813 patients was positive by either assay. Although no cases of acute infection were detected, 4% (11/272) of serum collected from a subset of patients was positive for LCMV-specific IgG. A significantly greater rate of seropositivity was found among subjects over 60 years of age (9.4%; P < 0.025) than was found in younger subjects (2.4% at 30-59 years of age; 0% at < 30 years of age). These data suggest that serious central nervous system disease due to LCMV infection is not common in this population. The high rate of seropositivity in those over 60 years of age suggest that infection was once more common.

Seroprevalence of lymphocytic choriomeningitis virus and Ljungan virus in Finnish patients with suspected neurological infections

“…LCMV nucleic acids were not detected in any of the CSF samples, and no LCMV seroconversion with IgM positivity was detected. This result is not unexpected, as LCMV does not appear to induce long viremia in humans . In previous studies, serum and CSF samples of patients hospitalized for meningitis, encephalitis, convulsions, and other CNS diseases were extensively screened for LCMV by RT‐PCR, but LCMV was not detected in any of the CSF samples over the study period, even though some of the samples were positive for LCMV IgG or IgM.…”

Multiple-Year Experience in the Diagnosis of Viral Central Nervous System Infections with a Panel of Polymerase Chain Reaction Assays for Detection of 11 Viruses

“…In contrast, there have been comparatively few reports of the application of PCR to relatively large numbers of patients with suspected viral CNS infections [5][6][7][8][9]. For several years, the Virology Laboratories at the New York State Department of Health's Wadsworth Center (Albany) have been using a battery of PCR assays designed for application to CSF and brain tissue to detect a range of viruses.…”

Strand-Specific Quantitative Reverse Transcription-Polymerase Chain Reaction Assay for Measurement of Arenavirus Genomic and Antigenomic RNAs

“…Several approaches have been utilized for the purpose of detecting and/or quantitating the different viral RNA species generated during LCMV infection. These approaches have ranged from standard RT-PCR to screen for the presence or absence of a particular viral RNA species [ 14 , 40 , 41 , 42 ] to Northern blot [ 11 ], in situ hybridization [ 43 , 44 ], or RNase protection assay [ 14 ] to more accurately determine the quantities of a given target RNA. Limitations of the latter three assays include the requirement for large quantities of RNA and/or radioactive reagents coupled with limited sensitivity and throughput.…”