title: Virological diagnosis of central nervous system infections by use of PCR coupled with mass spectrometry analysis of cerebrospinal fluid samples.
abstract: Viruses are the leading cause of central nervous system (CNS) infections, ahead of bacteria, parasites, and fungal agents. A rapid and comprehensive virologic diagnostic testing method is needed to improve the therapeutic management of hospitalized pediatric or adult patients. In this study, we assessed the clinical performance of PCR amplification coupled with electrospray ionization-time of flight mass spectrometry analysis (PCR-MS) for the diagnosis of viral CNS infections. Three hundred twenty-seven cerebrospinal fluid (CSF) samples prospectively tested by routine PCR assays between 2004 and 2012 in two university hospital centers (Toulouse and Reims, France) were retrospectively analyzed by PCR-MS analysis using primers targeted to adenovirus, human herpesviruses 1 to 8 (HHV-1 to -8), polyomaviruses BK and JC, parvovirus B19, and enteroviruses (EV). PCR-MS detected single or multiple virus infections in 190 (83%) of the 229 samples that tested positive by routine PCR analysis and in 10 (10.2%) of the 98 samples that tested negative. The PCR-MS results correlated well with herpes simplex virus 1 (HSV-1), varicella-zoster virus (VZV), and EV detection by routine PCR assays (kappa values [95% confidence intervals], 0.80 [0.69 to 0.92], 0.85 [0.71 to 0.98], and 0.84 [0.78 to 0.90], respectively), whereas a weak correlation was observed with Epstein-Barr virus (EBV) (0.34 [0.10 to 0.58]). Twenty-six coinfections and 16 instances of uncommon neurotropic viruses (HHV-7 [n = 13], parvovirus B19 [n = 2], and adenovirus [n = 1]) were identified by the PCR-MS analysis, whereas only 4 coinfections had been prospectively evidenced using routine PCR assays (P < 0.01). In conclusion, our results demonstrated that PCR-MS analysis is a valuable tool to identify common neurotropic viruses in CSF (with, however, limitations that were identified regarding EBV and EV detection) and may be of major interest in better understanding the clinical impact of multiple or neglected viral neurological infections.
authors.affiliation: Clinical and Molecular Virology Unit, University Hospital and EA-4684 Cardiovir SFR-CAP Santé, Faculty of Medicine, Reims, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Lévêque
authors.forename: Jérôme
authors.initials: J
authors.lastname: Legoff
authors.forename: Catherine
authors.initials: C
authors.lastname: Mengelle
authors.forename: Séverine
authors.initials: S
authors.lastname: Mercier-Delarue
authors.forename: Yohan
authors.initials: Y
authors.lastname: N'guyen
authors.forename: Fanny
authors.initials: F
authors.lastname: Renois
authors.forename: Fabien
authors.initials: F
authors.lastname: Tissier
authors.forename: François
authors.initials: F
authors.lastname: Simon
authors.forename: Jacques
authors.initials: J
authors.lastname: Izopet
authors.forename: Laurent
authors.initials: L
authors.lastname: Andréoletti
mesh_terms: D000046:Academic Medical Centers; D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002494:Central Nervous System Infections / Q000175:diagnosis* / Q000821:virology; D002555:Cerebrospinal Fluid / Q000821:virology*; D002648:Child; D002675:Child, Preschool; D005260:Female; D005602:France; D006785:Hospitals, University; D006801:Humans; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D014777:Virus Diseases / Q000175:diagnosis* / Q000821:virology; D014780:Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D055815:Young Adult