Detection of infection and determination of biomarkers for brucella melitensis infection in goats

Brucellosis is a chronic disease caused by Brucella spp. Brucella melitensis is one of the species of bacteria that infects animals and humans leading to undulant fever. The interaction between B. melitensis and immune system of the host could guide us towards the discovery of new biomarker to det...

Full description

Saved in:
Bibliographic Details
Main Author: Al-Garadi, Maged Ahmed Muthanna
Format: Thesis
Language:English
Published: 2011
Subjects:
Online Access:http://psasir.upm.edu.my/id/eprint/41846/1/FPV%202011%2015R.pdf
Tags: Add Tag
No Tags, Be the first to tag this record!
id my-upm-ir.41846
record_format uketd_dc
spelling my-upm-ir.418462016-03-03T09:22:42Z Detection of infection and determination of biomarkers for brucella melitensis infection in goats 2011-05 Al-Garadi, Maged Ahmed Muthanna Brucellosis is a chronic disease caused by Brucella spp. Brucella melitensis is one of the species of bacteria that infects animals and humans leading to undulant fever. The interaction between B. melitensis and immune system of the host could guide us towards the discovery of new biomarker to detect the infection as early as possible particularly understanding the unique pathway of B. melitensis and the intracellular features. In this study, 288 samples of whole blood and serum were collected from a goat farm in Kedah, Malaysia, which was suspected to have been affected by brucellosis. Serological and molecular detections of brucellosis were performed, using Rose Bengal Plate Test (RBPT), Complement Fixation Test (CFT), conventional PCR and real-time PCR (RT-PCR). Each test was evaluated with respect to its sensitivity and specificity ensuring that the recommendations made are able to be use for the national brucellosis eradication program in Malaysia. Isolation and identification of B. melitensis were also done in addition to conventional PCR and real-time PCR to detect B. melitensis from samples collected from vaginal swabs. CDs markers and its combination in different population of peripheral blood mononuclear cells (PMNC) were measured in detail, in different B. melitensis stages of the infection and in experimentally infected mice and goats by using specific monoclonal antibody. Histopathological picture was also been described in current study. The sensitivity of RBPT was 89.04% whilst CFT was 97.02%. The specificity of each of RBPT and CFT were 99.06% and 96.38% respectively. Four (4) B.melitensis isolates were isolated from 300 vaginal samples and all isolate belonged to B. melitensis biotype 1. The real-time PCR was the easier and safer method for the confirmation of brucellosis in goat’s population. The CDs biomarkers namely; CD14, CD4, CD25 markers were identified as good markers for the different stages of B. melitensis infection. A combination of a serological test namely RBPT and molecular technique, in particular real-time PCR based on the IS711 region of a hypothetical protein, showed promising results. This combination can be used to reduce the number of false positive results, which can cause severe economical loss during the implementation of proposed eradication programs. Brucellosis in goats Goats - Infection 2011-05 Thesis http://psasir.upm.edu.my/id/eprint/41846/ http://psasir.upm.edu.my/id/eprint/41846/1/FPV%202011%2015R.pdf application/pdf en public phd doctoral Universiti Putra Malaysia Brucellosis in goats Goats - Infection
institution Universiti Putra Malaysia
collection PSAS Institutional Repository
language English
topic Brucellosis in goats
Goats - Infection

spellingShingle Brucellosis in goats
Goats - Infection

Al-Garadi, Maged Ahmed Muthanna
Detection of infection and determination of biomarkers for brucella melitensis infection in goats
description Brucellosis is a chronic disease caused by Brucella spp. Brucella melitensis is one of the species of bacteria that infects animals and humans leading to undulant fever. The interaction between B. melitensis and immune system of the host could guide us towards the discovery of new biomarker to detect the infection as early as possible particularly understanding the unique pathway of B. melitensis and the intracellular features. In this study, 288 samples of whole blood and serum were collected from a goat farm in Kedah, Malaysia, which was suspected to have been affected by brucellosis. Serological and molecular detections of brucellosis were performed, using Rose Bengal Plate Test (RBPT), Complement Fixation Test (CFT), conventional PCR and real-time PCR (RT-PCR). Each test was evaluated with respect to its sensitivity and specificity ensuring that the recommendations made are able to be use for the national brucellosis eradication program in Malaysia. Isolation and identification of B. melitensis were also done in addition to conventional PCR and real-time PCR to detect B. melitensis from samples collected from vaginal swabs. CDs markers and its combination in different population of peripheral blood mononuclear cells (PMNC) were measured in detail, in different B. melitensis stages of the infection and in experimentally infected mice and goats by using specific monoclonal antibody. Histopathological picture was also been described in current study. The sensitivity of RBPT was 89.04% whilst CFT was 97.02%. The specificity of each of RBPT and CFT were 99.06% and 96.38% respectively. Four (4) B.melitensis isolates were isolated from 300 vaginal samples and all isolate belonged to B. melitensis biotype 1. The real-time PCR was the easier and safer method for the confirmation of brucellosis in goat’s population. The CDs biomarkers namely; CD14, CD4, CD25 markers were identified as good markers for the different stages of B. melitensis infection. A combination of a serological test namely RBPT and molecular technique, in particular real-time PCR based on the IS711 region of a hypothetical protein, showed promising results. This combination can be used to reduce the number of false positive results, which can cause severe economical loss during the implementation of proposed eradication programs.
format Thesis
qualification_name Doctor of Philosophy (PhD.)
qualification_level Doctorate
author Al-Garadi, Maged Ahmed Muthanna
author_facet Al-Garadi, Maged Ahmed Muthanna
author_sort Al-Garadi, Maged Ahmed Muthanna
title Detection of infection and determination of biomarkers for brucella melitensis infection in goats
title_short Detection of infection and determination of biomarkers for brucella melitensis infection in goats
title_full Detection of infection and determination of biomarkers for brucella melitensis infection in goats
title_fullStr Detection of infection and determination of biomarkers for brucella melitensis infection in goats
title_full_unstemmed Detection of infection and determination of biomarkers for brucella melitensis infection in goats
title_sort detection of infection and determination of biomarkers for brucella melitensis infection in goats
granting_institution Universiti Putra Malaysia
publishDate 2011
url http://psasir.upm.edu.my/id/eprint/41846/1/FPV%202011%2015R.pdf
_version_ 1747811892491976704