title: Analytical validation of a lymphocytic choriomeningitis virus real-time RT-PCR assay.
abstract: Lymphocytic choriomeningitis virus (LCMV) is a rare cause of central nervous system disease in humans. Screening by real-time RT-PCR assay is of interest in the case of aseptic meningitis of unknown etiology. A specific LCMV real-time RT-PCR assay, based on the detection of genomic sequences of the viral nucleoprotein (NP), was developed to assess the presence of LCMV in cerebrospinal fluids (CSF) sent for viral screening to a Swiss university hospital laboratory. A 10-fold dilution series assay using a plasmid containing the cDNA of the viral NP of the LCMV isolate Armstrong (Arm) 53b demonstrated the high sensitivity of the assay with a lowest detection limit of ≤50 copies per reaction. High sensitivity was confirmed by dilution series assays in a pool of human CSF using four different LCMV isolates (Arm53b, WE54, Traub and E350) with observed detection limits of ≤10PFU/ml (Arm53b and WE54) and 1PFU/ml (Traub and E350). Analysis of 130 CSF showed no cases of acute infection. The absence of positive cases was confirmed by a published PCR assay detecting all Old World arenaviruses. This study validates a specific and sensitive real-time RT-PCR assay for the diagnosis of LCMV infections. Results showed that LCMV infections are extremely rare in hospitalized patients western in Switzerland.
authors.affiliation: Swiss National Reference Centre for Emerging Viral Diseases, University of Geneva Hospitals, Geneva, Switzerland. samuel.cordey@hcuge.ch
authors.forename: Samuel
authors.initials: S
authors.lastname: Cordey
authors.forename: Roland
authors.initials: R
authors.lastname: Sahli
authors.forename: Marie-Laurence
authors.initials: ML
authors.lastname: Moraz
authors.forename: Christine
authors.initials: C
authors.lastname: Estrade
authors.forename: Laurence
authors.initials: L
authors.lastname: Morandi
authors.forename: Pascal
authors.initials: P
authors.lastname: Cherpillod
authors.forename: Remi N
authors.initials: RN
authors.lastname: Charrel
authors.forename: Stefan
authors.initials: S
authors.lastname: Kunz
authors.forename: Laurent
authors.initials: L
authors.lastname: Kaiser
mesh_terms: D000595:Amino Acid Sequence; D001117:Arenaviridae Infections / Q000175:diagnosis*; D001483:Base Sequence; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007223:Infant; D008217:Lymphocytic choriomeningitis virus / Q000235:genetics / Q000302:isolation & purification*; D008297:Male; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins / Q000235:genetics; D010802:Phylogeny; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D013557:Switzerland
chemical_list: D019590:Nucleocapsid Proteins