Cycling probe technology to quantify and discriminate between wild-type varicella-zoster virus and Oka vaccine strains

Faculty of Clinical Engineering, Fujita Health University School of Health Sciences, Toyoake, Aichi, Japan. Electronic address: .
Journal of virological methods (Impact Factor: 1.78). 06/2013; 193(2). DOI: 10.1016/j.jviromet.2013.06.031
Source: PubMed


Rapid differentiation between wild-type varicella zoster virus (VZV) and Oka-vaccine (vOka) strains is important for monitoring side reactions of varicella vaccination. To develop a high-throughput molecular diagnostic method for the differentiation of wild-type VZV and vOka strains based on cycling probe technology. The primers were designed to amplify common sequences spanning a single nucleotide polymorphism (SNP) in gene 62 of VZV. DNA-RNA chimeric probes (cycling probes) were designed to detect the SNP at nucleotide 105705. The cycling probe real-time PCR assays for VZV wild-type and vOka strains specifically amplified plasmids containing target sequences that ranged between 10 and 1×10(6) copies per reaction. The inter- and intra-assay coefficients of variation were less than 5%. After initial validation studies, the clinical reliability of this method was evaluated using 38 swab samples that were collected from patients suspected of being zoster. Compared to the loop mediated isothermal amplification method, which is defined as the gold standard, cycling probe real-time PCR was highly sensitive and specific. The cycling probe real-time PCR technology is a reliable tool for differentiating between wild-type VZV and vOka strains in clinical samples.

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Available from: Yoshiki Kawamura, Sep 22, 2015
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    ABSTRACT: HSV and varicella zoster virus (VZV) are common pathogens of skin and mucous membranes and the CNS. Their rapid and accurate diagnosis is essential for their treatment as well as infection control. Cytological and morphological examination, specific antibody detection methods and virus isolation all have their own limitations in clinical practice. In recent years, molecular methods have become the primary diagnostic methods for the detection and differentiation of HSV1/2 and VZV due to their high sensitivity, specificity, rapid test turnaround time and potential for high throughput and automation. Although molecular assays detect HSV1/2 and VZV more quickly, the clinical significance of positive results may vary in individual patients.
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