Key result
A novel multiplex RT-LAMP assay reliably detected and discriminated between FMDV, SVDV, and VSNJV in epithelial suspensions without prior RNA extraction, with sensitivity equivalent to RT-PCR.
The novel multiplex RT-LAMP assay provides a rapid, low-cost method for differentiating foot-and-mouth disease from other vesicular diseases in the field without prior RNA extraction.
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May simplify field diagnostics for vesicular viruses; leaves open prospective validation before clinical adoption.
Vesicular stomatitis (VS) is endemic in Central America and northern regions of South America, where sporadic outbreaks in cattle and pigs can cause clinical signs that are similar to foot-and-mouth disease (FMD). There is therefore a pressing need for rapid, sensitive and specific differential diagnostic assays that are suitable for decision making in the field. RT-LAMP assays have been developed for vesicular diseases such as FMD and swine vesicular disease (SVD) but there is currently no RT-LAMP assay that can detect VS virus (VSV), nor are there any multiplex RT-LAMP assays which permit rapid discrimination between these 'look-a-like' diseases in situ. This study describes the development of a novel RT-LAMP assay for the detection of VSV focusing on the New Jersey (VSNJ) serotype, which has caused most of the recent VS cases in the Americas. This RT-LAMP assay was combined in a multiplex format combining molecular lateral-flow devices for the discrimination between FMD and VS. This assay was able to detect representative VSNJV's and the limit of detection of the singleplex and multiplex VSNJV RT-LAMP assays were equivalent to laboratory based real-time RT-PCR assays. A similar multiplex RT-LAMP assay was developed to discriminate between FMDV and SVDV, showing that FMDV, SVDV and VSNJV could be reliably detected within epithelial suspensions without the need for prior RNA extraction, providing an approach that could be used as the basis for a rapid and low cost assay for differentiation of FMD from other vesicular diseases in the field.
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Fowler et al. (2016) studied Vesicular stomatitis and Foot-and-mouth disease. Multiplex RT-LAMP assay vs. Real-time RT-PCR assays was evaluated on Limit of detection and discrimination between vesicular disease viruses. A novel multiplex RT-LAMP assay reliably detected and discriminated between FMDV, SVDV, and VSNJV in epithelial suspensions without prior RNA extraction, with sensitivity equivalent to RT-PCR.
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