Infectious Bursal disease

To investigate: One-step reverse-transcription loop-mediated isothermal amplification for detection of infectious bursal disease virus

• the researchers here used a one-step RT-LAMP assay to detect IBDV RNA genome. While LAMP typically does not work well for RNA viruses this assay combines the LAMP method with the use of AMV RTase which is known to have a broader range of reaction temperatures making it more optimal then the traditional use of M-MLV RTase. This also allows for specific detection of IBDV infection in broiler chickens. The researcher only used 4 primers which are available in the paper (typical LAMP assays use between 4 and 6 primers.) The optimized conditions are 60°C and a reaction time as short as 40 minutes which makes it a great choice for quick field assessment. Additionally, you are able to visualize the results with either fluorescence or the turbidity of the solution. Their researcher proved that the RT-LAMP method is both highly sensitive and accurate.

Khan RSA, Ali W, Kiran S, Shah MSD, Tahir ZA, Habib M. (2018). Rapid detection of infectious bursal disease by loop-mediated isothermal amplification for field analysis.

• The researchers compared RT-LAMP against RT-PCR as detection methods for IBD. Their results showed that RT-LAMP is found to have 10 times higher selectivity for IBDV when compared to RT-LAMP and was able to detect 9.2% more field samples than by RT-PCR. They were able to declare a field sample positive based on the turbidity or by adding fluorescence staining reagent like SYBR green I. they created 6 primers for RT-LAMP which were specific for the VP5 gene (two inner and two outer primers as well as loop primers). They additionally provide the primers they used for RT-PCR. There was no need for a thermos cycler or other heavy instruments with RT-LAMP and a fluorescent dsDNA intercalating dye was used for visual detection.