Standardization of Loop mediated isothermal amplification for detection of D. nodosus and F. necrophorum causing footrot in sheep and goats

The Loop Mediated Isothermal Amplification (LAMP) was standardized for rapid detection of D. nodosus and F. necrophorum . A total of 250 foot swabs (200) were screened from sheep and (50) were from goats from different districts of Rayalaseema viz., Chittoor, Nellore, Kadapa, Anantapur. Out of 250 samples 75 (30.0%) and 85 (34.0%) were positive for D. nodosus and F. necrophorum respectively. All the 250 samples were screened individually for both the organisms by LAMP. Among them, 104 (41.6%) were found to be positive for D. nodosus and 120 (48.0%) were positive for F. necrophorum . The efficacy of LAMP in terms of sample DNA detection limit was compared with the PCR by using standard dilutions of DNA extracted from D. nodosus and F. necrophorum cultures. The detection limit was found to be higher than PCR for both the organisms. The sensitivity of LAMP is compared with PCR by targeting 16S rRNA gene of D. nodosus and lktA gene of F. necrophorum. In case of D. nodosus , out of 250 samples, 75 (30.0%) were positive by PCR and 104 (41.6%) were positive by LAMP. Among 250 samples, 85 (34.0%) were positive by PCR and 120 (48.0%) were positive by LAMP in case of F. necrophorum . The LAMP was found to be more sensitive than PCR in detecting the organisms with high statistical significance.