Blue Tongue

Reverse-transcription-loop-mediated-isothermal-amplification (RT-LAMP) assay were created to detect eastern or western topotype of BTV. They created primers that recognized 6-distinct sequences of BTV genome segment 1. These assays were able to detect BTV RNA in all positive isolates tested. The sensitivity of RT-LAMP is similar to the sensitivity of RT-PCR but higher than conventional RT-PCR. Could not detect other local viruses that cause symptoms similar to BTV.

using RT-PCR the researchers have created a one-step single-tube multiplex RT-PCR that is able to detect not only BTV but RVFV, RPV and PPRV. Using samples of whole blood or oral mucosal tissue they were able to detect these viruses. This multiplex RT-PCR had sensitivity that was 10-fold lower than that of the corresponding uniplex RT-PCR in oral mucosal homogenate suspension than in whole blood. would be a great choice for doing screen testing on animal in an outbreak because whole blood would typically be used over oral mucosal. much more cost effective than uniplex assays.

Karthika Lakshmi, Kalyani Putty, Satya Samparna Raut, Sunil R. Patil, P. P. Rao, B. Bhagyalakshmi, Y. Krishna Jyothi, B. Susmitha, Y. Vishnuvardhan Reddy, Sowmya Kasulanati, J. Shiva Jyothi and Y. N. Reddy. (2018) Standardization and application of real-time polymerase chain reaction for rapid detection of bluetongue virus

Karthika et al standardized real-time PCR for detecting BTV from blood samples collected during an outbreak in India in 2014. They found that real-time PCR had a higher sensitivity then compared to rt-PCR when used for field detection. In order to create the proper primers for they targeted NS3 which is reliable for BTV virulence. At present, only two of the 11 segments coded in the genome are responsible for serotype and are succeptible to recombination so targeting other structural proteins allow for field detection of almost all serotypes

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