title: Molecular detection of varicella zoster virus while keeping an eye on the budget.
abstract: Varicella zoster virus (VZV) PCR is highly sensitive compared to traditional detection methods like culture and direct fluorescent antibody testing (DFA); however, the high cost of commercial assays prohibits their use in many clinical laboratories. Major contributors to cost are the nucleic acid extraction and the PCR reagents. This study evaluated an "in-house" qualitative real-time PCR where the nucleic acid extraction was replaced by a crude extraction, homogenization and heat treatment. Three methods were compared: virus culture and DFA and real-time PCR following each extraction methods. The real-time PCR was highly specific for VZV, and the analytical sensitivity was equivalent following both extraction methods. In contrast, virus culture and DFA was approximately 10,000-fold less sensitive. Using 200 clinical specimens, the sensitivity for the real-time PCR following nucleic acid extraction or homogenization and heat treatment was essentially equivalent at 100% and 97.2%, respectively; whereas, virus culture and DFA was significantly less sensitive at 54.8%. Overall, homogenization and heat treatment combined with a qualitative in-house real-time PCR is a rapid, accurate and cost effective method for the detection of VZV.
authors.affiliation: Department of Pathology and Laboratory Medicine, Capital District Health Authority, Halifax, Nova Scotia, Canada; Dalhousie University, Halifax, Nova Scotia, Canada.
authors.forename: Khalifa
authors.initials: K
authors.lastname: Binkhamis
authors.affiliation: Department of Pathology and Laboratory Medicine, Capital District Health Authority, Halifax, Nova Scotia, Canada; Dalhousie University, Halifax, Nova Scotia, Canada.
authors.forename: Turkiya
authors.initials: T
authors.lastname: Al-Siyabi
authors.affiliation: Department of Pathology and Laboratory Medicine, Capital District Health Authority, Halifax, Nova Scotia, Canada.
authors.forename: Charles
authors.initials: C
authors.lastname: Heinstein
authors.affiliation: Department of Pathology and Laboratory Medicine, Capital District Health Authority, Halifax, Nova Scotia, Canada; Dalhousie University, Halifax, Nova Scotia, Canada.
authors.forename: Todd F
authors.initials: TF
authors.lastname: Hatchette
authors.affiliation: Department of Pathology and Laboratory Medicine, Capital District Health Authority, Halifax, Nova Scotia, Canada; Dalhousie University, Halifax, Nova Scotia, Canada. Electronic address: jason.leblanc@cdha.nshealth.ca.
authors.forename: Jason J
authors.initials: JJ
authors.lastname: LeBlanc
mesh_terms: D002644:Chickenpox / Q000175:diagnosis* / Q000821:virology; D003365:Costs and Cost Analysis; D004279:DNA, Viral / Q000302:isolation & purification; D017048:Health Care Costs; D006562:Herpes Zoster / Q000175:diagnosis* / Q000821:virology; D014645:Herpesvirus 3, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000191:economics* / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000191:economics* / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral
keywords: Cost analysis; Homogenization; Nucleic acid extraction; Real-time PCR; Varicella zoster virus