A Quantitative Real-Time RT-PCR Assay for the Detection of Venezuelan equine encephalitis virus Utilizing a Universal Alphavirus Control RNA

Figure 1

The nucleotide sequence of the synthetic construct used for calibration of the EEV-specific qRT-PCRs (a). The target sequences (underlined) for the specific qRT-PCRs are cloned into the vector pCR2.1-TOPO. Within the VEEV target region a modified sequence (framed) allows the differentiation of the synthetic calibrator from viral sequences. Nucleotide exchanges are indicated in red. Amplification curves of the qRT-PCRs specific for VEEV (b), EEEV (c), and WEEV (d) using the synthetic calibrator template. Amplification curve of the synthetic calibrator targeted with the control probe is indicated in olive, respectively. Standard curves (see enclosed boxed figures) were obtained by Ct values plotted against the log of the starting quantity. Calculated correlation coefficients (), slopes, and amplification efficiencies () are depicted below the corresponding figures.

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