Diagnosis of Barmah Forest Virus Infection by a Nested Real-Time SYBR Green RT-PCR Assay |
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| Authors: | Linda Hueston Cheryl S. Toi Neisha Jeoffreys Tania Sorrell Gwendolyn Gilbert |
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| Affiliation: | 1. Centre for Infectious Diseases and Microbiology Laboratory Services, Westmead, NSW, Australia.; 2. Sydney Institute for Emerging Infectious Diseases and Biosecurity, Westmead, NSW, Australia.; University of Hong Kong, Hong Kong, |
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| Abstract: | Barmah Forest virus (BFV) is a mosquito borne (+) ssRNA alphavirus found only in Australia. It causes rash, myalgia and arthralgia in humans and is usually diagnosed serologically. We developed a real-time PCR assay to detect BFV in an effort to improve diagnosis early in the course of infection. The limit of detection was 16 genome equivalents with a specificity of 100%. Fifty five serum samples from BFV-infected patients were tested by the PCR. 52 of 53 antibody-positive samples were PCR negative. Two culture-positive (neutralizing antibody negative) samples were positive on first round PCR, while one sample (IgM and neutralizing antibody strongly positive, IgG negative) was positive on second round PCR, suggesting that viral RNA is detectable and transiently present in early infection. PCR can provide results faster than culture, is capable of high throughput and by sequencing the PCR product strain variants can be characterized. |
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