DEVELOPMENT OF IN-HOUSE ELISA FOR THE DETECTION OF Mycobacterium avium subspecies paratuberculosis INFECTION IN ANIMALS
| dc.contributor.advisor | Dr. D. RATHNAMMA | |
| dc.contributor.author | PANNAGA, P. | |
| dc.date.accessioned | 2021-01-21T09:34:24Z | |
| dc.date.available | 2021-01-21T09:34:24Z | |
| dc.date.issued | 2017-10 | |
| dc.description.abstract | The present study was under taken to develop an in-house ELISA using a local Mycobacterium avium subspecies paratuberculosis (MAP) isolate and to investigate the seroprevalance of Paratuberculosis in animals. The MAP protoplasmic antigen was extracted by sonication and quantified by ‘Nanodrop’ spectrophotometer and 4.193 mg/ml antigen was obtained. Extracted MAP antigen was characterized by SDS - PAGE and immune-blot analysis. The SDS-PAGE profile of MAP revealed 25, 35, 57, 72, 100 and 193 kDa proteins and immunoblotting revealed that 20, 29, 35, 45, 70 and 193 kDa as immunogenic proteins. ELISA was standardized with MAP antigen of 0.125 μg / well, serum dilution of 1:50 and Protein A-HRP conjugate dilution of 1:2500. 1032 serum samples from sheep, goat and cattle were screened for MAP infection with in-house ELISA and IDEXX ELISA kit. 113 (10.94 %) serum samples were positive by in-house ELISA and 111 (10.75 %) serum samples were positive by IDEXX ELISA kit. Cut off values were determined as SP ratio ≥ 80 per cent as Positive, ≤ 60 per cent as Negative and 60 - 80 per cent as suspected. Seroprevalence of MAP in sheep, goat and cattle was 8.52, 5.95 and 6.6 per cent respectively. The relative sensitivity and specificity of in house ELISA was 76.58 per cent and 96.96 per cent respectively with Kappa value of 0.7296 showing good agreement. Comparative evaluation of in-house ELISA with IDEXX ELISA kit was statistically analysed by Pearson’s correlation with significant correlation of 0.7725 (P=0.0001) between in-house ELISA and IDEXX ELISA kit. Receiver operating characteristics (ROC) curve was drawn with excellent area under curve of 0.992 was obtained. The study indicated the superiority of in-house indirect ELISA using native MAP antigen as against purified protoplasmic MAP antigens used in commercial ELISA kit in terms of cost effectiveness. | en_US |
| dc.identifier.uri | https://krishikosh.egranth.ac.in/handle/1/5810160497 | |
| dc.keywords | VETERINARY MICROBIOLOGY | en_US |
| dc.language.iso | English | en_US |
| dc.pages | 152 | en_US |
| dc.publisher | KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCIES UNIVERSITY, BIDAR | en_US |
| dc.sub | Veterinary Microbiology | en_US |
| dc.theme | VETERINARY MICROBIOLOGY | en_US |
| dc.these.type | M.V.Sc. | en_US |
| dc.title | DEVELOPMENT OF IN-HOUSE ELISA FOR THE DETECTION OF Mycobacterium avium subspecies paratuberculosis INFECTION IN ANIMALS | en_US |
| dc.type | Thesis | en_US |