title: Development of a multiplex real-time PCR for the simultaneous detection of herpes simplex and varicella zoster viruses in cerebrospinal fluid and lesion swab specimens.
abstract: Herpes simplex viruses (HSV) and varicella zoster virus (VZV) can have very similar and wide-ranging clinical presentations. Rapid identification is necessary for timely antiviral therapy, especially with infections involving the central nervous system, neonates, and immunocompromised individuals. Detection of HSV-1, HSV-2 and VZV was combined into one real-time PCR reaction utilizing hydrolysis probes. The assay was validated on the LightCycler(®) (Roche) and Applied Biosystems 7500 Real-Time PCR System (Thermo Fisher Scientific Inc.) to detect alphaherpesviruses in cerebral spinal fluid (CSF) and lesion swab specimens, respectively. Validation data on blood and tissue samples are also presented. The multiplex assay showed excellent sensitivity, specificity and reproducibility when compared to two singleplex real-time PCR assays for CSF samples and direct fluorescent antigen/culture for lesion swab samples. Implementation of the multiplex assay has facilitated improved sensitivity and accuracy as well as reduced turn-around-times and costs. The results from a large data set of 16,622 prospective samples tested between August 16, 2012 to February 1, 2014 at the Provincial Laboratory for Public Health (Alberta, Canada) are presented here.
authors.affiliation: Provincial Laboratory for Public Health (Microbiology), 3030 Hospital Drive NW, Calgary, Alberta, Canada T2N 4W4.
authors.forename: Anita A
authors.initials: AA
authors.lastname: Wong
authors.affiliation: Provincial Laboratory for Public Health (Microbiology), 3030 Hospital Drive NW, Calgary, Alberta, Canada T2N 4W4. Electronic address: Kanti.Pabbaraju@albertahealthservices.ca.
authors.forename: Kanti
authors.initials: K
authors.lastname: Pabbaraju
authors.affiliation: Provincial Laboratory for Public Health (Microbiology), 3030 Hospital Drive NW, Calgary, Alberta, Canada T2N 4W4.
authors.forename: Sallene
authors.initials: S
authors.lastname: Wong
authors.affiliation: Provincial Laboratory for Public Health (Microbiology), 3030 Hospital Drive NW, Calgary, Alberta, Canada T2N 4W4; Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Raymond
authors.initials: R
authors.lastname: Tellier
mesh_terms: D000416:Alberta; D002170:Canada; D002555:Cerebrospinal Fluid / Q000821:virology*; D002644:Chickenpox / Q000175:diagnosis; D018792:Encephalitis, Viral / Q000175:diagnosis; D006561:Herpes Simplex / Q000175:diagnosis; D014645:Herpesvirus 3, Human / Q000302:isolation & purification*; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D011446:Prospective Studies; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D018139:Simplexvirus / Q000302:isolation & purification*; D012867:Skin / Q000821:virology*
keywords: Cerebral-spinal fluid; Herpes simplex virus; Lesion; Multiplex; Real-time PCR; Varicella zoster virus