I=
Pakistan Research Repository Home
 

Title of Thesis

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

Author(s)

Syed Ehtisham ul Haque

Institute/University/Department Details
Department Of Microbiology, Faculty Of Veterinary Science / University Of Agriculture, Faisalabad
Session
2010
Subject
Veterinary Microbiology
Number of Pages
235
Keywords (Extracted from title, table of contents and abstract of thesis)
Molecular, Mycoplasma, Diagnostic, Epidemiological, Poultry, Synoviae, Gallisepticum, Investigations, Diagnostic, Sequence, Pathogens

Abstract
Not Available

Download Full Thesis
2,965 KB
S. No. Chapter Title of the Chapters Page Size (KB)
1 0 CONTENTS

 

 
25 KB
2

1

INTRODUCTION

 

1
133 KB
3 2 REVIEW OF LITERATURE

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

7
269 KB
4 3 MATERIALS AND METHODS

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

35
 250 KB
5 4 RESULTS

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

59
2,430 KB
6 5 DISCUSSION

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

171
154 KB
7 6 SUMMARY

 

184
126 KB
8 7 LITERATURE CITED AND APPENDICES

189
208 KB