Mhase, P. P.Vanjari, A. D.2023-03-162023-03-162021https://krishikosh.egranth.ac.in/handle/1/5810195299The present investigation was undertaken in periphery of hundred kilometers around Shirwal with aim of detection of M. gallisepticum infection in poultry. Out of 106 tissue samples tested by PCR, 7 (6.60 %) were positive for M. gallisepticum, whereas all 24 choanal swab samples were negative. Four flocks out 11 were positive for M. gallisepticum infection. Flocks from Bhor and Wai area were positive for M. gallisepticum by specific PCR on the pooled tissues. PCR positive samples were used for isolation of M. gallisepticum in view of their confirmation with gold standard. However, M. gallisepticum was culturally isolated from 05/99 (5.05 %) of tissue samples. Sequence analysis of PCR amplicons from standard control (M. gallisepticum vaccine strain DNA) and field sample was carried out using SeqMan DNASTAR and NCBI BLAST. Blast analysis of field sample showed highest alignment with PB1/06/Ind. The novel PSR technique with different gradient combinations using 16SrRNA and lipoprotein gene primer sets were attempted with different gradient combinations of temperature (60 ºC, 61 ºC , 62 ºC , 63 ºC, 64 ºC, 65 ºC); extension time (20 min, 30min, 40min, 50 min, 60 min, 120 min for time) and MgSO4 (2,3,4,5,6 mM) concentrations. But, none of the tested protocols yielded positive results, hence, primer sets and reaction conditions used in this study were not found optimum for PSR detection of M. gallisepticum.EnglishThesis