title: Evaluation of a multiplex polymerase chain reaction for early diagnosis of ventriculostomy-related infections.
abstract: OBJECTIVE
  Diagnosis of ventriculostomy-related infections (VRIs) is challenging due to the lack of rapid, sensitive assays for pathogen detection. The authors report the development of a multiplex polymerase chain reaction (PCR) assay for differential diagnosis of common VRI pathogens.
  
  
  METHODS
  MassTag PCR was used to develop a multiplex assay for detection of 11 VRI pathogens. The assay was established and optimized using cloned template standards and spiked samples and was then evaluated on CSF specimens from ventricular drains. Subjects were grouped into definite VRI, possible VRI, or no VRI based on conventional microbiology, CSF evaluation, and clinical parameters.
  
  
  RESULTS
  CSF specimens were obtained from 45 subjects (median age 49 years, interquartile range 32-63 years; 51% were male). The assay detected 10-100 genome copies. It detected a pathogen in 100% (6 of 6) of definite VRI cases in which a pathogen targeted by the assay was present; these represented 67% of all definite VRIs (6 of 9). Among subjects with a possible VRI, the assay detected a pathogen in 29% (5 of 17). In subjects without overt infection the presence of a pathogen was detected in 32% of subjects (6 of 19), albeit with lower signal compared with the VRI group.
  
  
  CONCLUSIONS
  MassTag PCR enabled parallel testing of CSF specimens for 11 pathogens of VRI. The high sensitivity of PCR combined with possible device colonization, specimen contamination, and concurrent antibiotic treatments limit the clinical value of the assay, similar to other current diagnostic approaches. With further optimization, multiplex PCR may provide timely identification of multiple possible VRI pathogens and guide management, complementing classic culture approaches.
authors.affiliation: Division of Infectious Disease, Department of Medicine, and.|Department of Medicine, University of Melbourne, Victoria, Australia.
authors.forename: Claire L
authors.initials: CL
authors.lastname: Gordon
authors.affiliation: Center for Infection and Immunity and.
authors.forename: Rafal
authors.initials: R
authors.lastname: Tokarz
authors.affiliation: Center for Infection and Immunity and.|Department of Epidemiology, Mailman School of Public Health, Columbia University Medical Center;
authors.forename: Thomas
authors.initials: T
authors.lastname: Briese
authors.affiliation: Center for Infection and Immunity and.
authors.forename: W Ian
authors.initials: WI
authors.lastname: Lipkin
authors.affiliation: Center for Infection and Immunity and.
authors.forename: Komal
authors.initials: K
authors.lastname: Jain
authors.affiliation: Clinical Microbiology Service, NewYork-Presbyterian Hospital, New York, New York; and.
authors.forename: Susan
authors.initials: S
authors.lastname: Whittier
authors.affiliation: Division of Infectious Disease, Department of Medicine, and.
authors.forename: Jayesh
authors.initials: J
authors.lastname: Shah
authors.affiliation: Department of Neurosurgery, Columbia University Medical Center;
authors.forename: E Sander
authors.initials: ES
authors.lastname: Connolly
authors.affiliation: Division of Infectious Disease, Department of Medicine, and.
authors.forename: Michael T
authors.initials: MT
authors.lastname: Yin
mesh_terms: D000328:Adult; D002176:Candida albicans / Q000302:isolation & purification*; D058565:Cerebral Ventriculitis / Q000175:diagnosis / Q000382:microbiology*; D002555:Cerebrospinal Fluid / Q000382:microbiology; D042241:Early Diagnosis; D005260:Female; D006090:Gram-Negative Bacteria / Q000302:isolation & purification*; D006094:Gram-Positive Bacteria / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D008875:Middle Aged; D060885:Multiplex Polymerase Chain Reaction; D014696:Ventriculostomy / Q000009:adverse effects*
keywords: CoNS = coagulase-negative Staphylococcus spp.; MRSA = methicillin-resistant Staphylococcus aureus; MSSA = methicillin-sensitive S. aureus; NYP = NewYork-Presbyterian; PCR; PCR = polymerase chain reaction; VRI = ventriculostomy-related infection; diagnosis; ventriculostomy infection