Molecular Detection of Mycoplasma gallisepticum in Chicken
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Date
2021
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MAFSU, Nagpur
Abstract
The 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.