Title of Thesis

Diagnostic And Molecular Epidemiological Investigations Of Mycoplasma Gallisepticum And Mycoplasma Synoviae In Poultry

Author(s)

Syed Ehtisham ul Haque

Abstract
Not Available

1

2.1 Avian Mycoplasmas
2.2 Serology and Mycoplasma clean programs
2.3 Mycoplasma PCR
2.4 16SrRNA PCR
2.5 mgc2 gene based PCR for Mycoplasma gallisepticum
2.6 Gene specific PCR and gene-targeted sequencing (GTS) for M. gallisepticum
2.7 vlhA gene based PCR for Mycoplasma synoviae
2.8 PvpA gene based PCR
2.9 Environmental detection of avian Mycoplasmas
2.10 Multiplex PCR for Mycoplasmas
2.11 Real-time PCR for Mycoplasma species
2.12 Multiplex real-time PCR
2.13 Mycoplasma genome sequence
2.14 Emerging avian Mycoplasma spp

3.1 Collection of samples
3.2 Serological screening
3.3 Clinical samples
3.4 Environmental samples
3.5 Avian Mycoplasma Reference cultures
3.6 Culture isolation
3.7 Isolation of microorganism
3.8 Identification of isolates
3.9 Biochemical characterization
3.10 RSA test antigen preparation
3.11 Polymerase chain reaction (PCR)
3.12 Preparation of PCR primers
3.13 PCR procedures
3.14 Thermal cycling
3.15 Electrophoresis
3.16 DNA sequencing
3.17 Real time PCR
3.18 TaqMan real time PCR
3.19 TaqMan real time PCR reaction mixture for Plus Minus TaqMan assay
3.20 Real time PCR reaction mixture preparation
3.21 Cloning of real time PCR product
3.22 Plasmid DNA Purification protocol

4.1 Breed wise distribution of samples positive for M. gallisepticum and M. synoviae cases screened through RSA and identified through Culture and PCR in broiler, layer and breeder flocks
4..2 Comparative flock wise prevalence of MG and MS at various level of sero-positivity
4.3 Comparative sample wise success rate of PCR from culture positive and negative samples for MG and MS
4.4 Environmental detection of avian Mycoplasma species
4.5 Mycoplasma DNA extraction from indigenous isolates
4.6 Optimization of 16SrRNA based PCR for MG and MS
4.7 Optimization of mgc2 based PCR for M. gallisepticum
4.8 Optimization of vlhA gene based PCR for MS
4.9 DNA sequencing of mgc2 and vlhA gene products
4.10 Development and optimization of real-time PCR for MG
4.11 Plus Minus assay with MG TaqMan MGB probe
4.12 Development and optimization of real time PCR for MS
4.13 Development and optimization of TaqMan based Plus Minus real-time PCR assay for MS
4.14 Development and optimization of TaqMan MGB duplex Real time PCR for MG and MS
4.15 Standard curve and quantitative detection of MG and MS through real-time PCR

5.1 Serology, culture and PCR for avian mycoplasmas
5.2 Conventional PCR
5.3 Efficacy of real-time PCR for the detection of M.gallisepticum
5.4 Efficacy of real-time PCR for the detection of M.synoviae
5.5 Standard Curve derivation for quantitative analysis of MG and MS through TaqMan based real-time PCR
5.6 Development and optimization of duplex Real-time PCR for simultaneous detection of MG and MS
5.7 Cloning of Real-time PCR product