Loop–Mediated Isothermal Amplification (LAMP) assay for rapid detection of Orf virus infection in sheep and goats in Karnataka.

Srinivasa Babu; T; D.Rathnamma; Shrikrishna Isloor; B.M.Chandranaik; H.D.Narayanaswamy; B.M.Veeregowda

Volume : VII, Issue : VIII, August - 2017

Loop–Mediated Isothermal Amplification (LAMP) assay for rapid detection of Orf virus infection in sheep and goats in Karnataka.

Srinivasa Babu, T, D. Rathnamma, Shrikrishna Isloor, B. M. Chandranaik, H. D. Narayanaswamy, B. M. Veeregowda

Abstract :

The Loop mediated isothermal amplification (LAMP) assay was standardized using six primers to amplify B2L gene of Orf virus (ORFV). Conventional polymerase chain reaction (PCR) was also standardized targeting B2L gene for detection of ORFV. A total of 70 scab samples were collected from sheep and goats showing symptoms of Orf disease. The PCR screening of 70 scab samples revealed 60 percent positivity for ORFV. The LAMP assay was standardized using Hydroxynapthol blue dye at the final working concentration of 200M in the reaction mixture for the visual detection of ORFV in PCR positive scab samples. The successful amplification was indicated by a colour change from deep blue to light blue. All the PCR positive samples were also found positive with LAMP assay. Although the LAMP reaction had equal sensitivity to the PCR test, it is considered superior because of its simplicity to carryout in field conditions and at quarantine inspection places.