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.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Binkhamis K,Al-Siyabi T,Heinstein C,Hatchette TF,LeBlanc JJdoi
10.1016/j.jviromet.2014.02.009subject
Has Abstractpub_date
2014-06-01 00:00:00pages
24-7eissn
0166-0934issn
1879-0984pii
S0166-0934(14)00052-4journal_volume
202pub_type
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