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Enzyme immunotransfer blot of different Brucella antigens against crude hyperimmune serum prepared in guinea pigs (Brucella S99 antiserum).
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kills Brucella cells by causing lysis of the membrane, so the phenol-heat killed brucella antigen may lake specificity as a result of destruction the majority of proteins in the cell wall. Accordingly, attention was directed to produce antigen using binary ethylene imine as an inactivator. The produced antigen showed high specificity in detecting B...
Citations
... Polymerase chain reaction is becoming very useful and considerable progress has been made to improve their sensitivity, specificity, and technical case and to lower costs. Nucleic acid amplification has been explored for rapid detection and confirmation of the presence of Brucella species [19]. ...
assessment of phenotype and host preference. Each year half a
million case of brucellosis occurs in humans around the world. Five
out of nine known Brucella species can infect humans. The most
pathogenic and invasive species for human are B. melitensis, B.
abortus and B. canis. Understanding the Brucella species and bio
variant through advance knowledge of molecular epidemiology has
signifiant role in the elucidating the source of infection, disease
transmission pattern and furthermore in designing specifi control
strategies through utilization of relevant vaccine in affected livestock
population. Prevention and control of brucellosis can be adopted
realistically through understanding of local and regional variations
in animal husbandry practices, social customs, infrastructures and
epidemiological patterns of the disease and species of Brucella.
Hence, this seminar paper attempted to highlight the molecular epidemiology and public health signifiance of brucellosis in livestock
and human populations.
Keywords: Brucellosis; Diagnosis; Livestock; Molecular epidemi
... Polymerase chain reaction is becoming very useful and considerable progress has been made to improve their sensitivity, specificity, and technical case and to lower costs. Nucleic acid amplification has been explored for rapid detection and confirmation of the presence of Brucella species [19]. ...
... unpractical to apply on a large scale in control campaigns. Accordingly, the indirect diagnosis of disease based on serological tests is the method of choice in the eradication programs (Hussein and Awad, 2007). ...
Brucellosis is an important disease affecting mainly sheep and goats. Diagnosis based on isolation of Brucella organisms from the suspected animals is the golden standard but has a limited sensitivity, expensive and unpractical to apply on a large scale in control campaigns. Accordingly, the indirect diagnosis of disease based on serological tests is the method of choice in the eradication programs. In this study, a single step polymerase chain reaction (PCR) was used to diagnose brucellosis using sheep whole blood and compared its sensitivity and specificity against some of the most commonly used serological techniques and modified ones. Three hundred apparently healthy ewes were randomly chosen from different governorates of Egypt. Sera were tested against Rose Bengal test (RBT), Serum Agglutination test (SAT), ELISA using both the whole Brucella antigen (W-ELISA) and the periplasmic protein antigen (P-ELISA). Results showed that 39% of the blood samples were positive to the PCR test, Meanwhile 29.3, 27.0, 28.7 and 28.3% were positive to the previous serological tests respectively. We recommend the use of this blood PCR assay for accurate diagnosis of ovine brucellosis especially in the early stage of infection, which is difficult to achieve by the applied serological tests.
Brucellosis is one of the most important diseases affecting human and animals in most of the developing countries including Egypt. Rapid and accurate diagnosis is fundamental for control and eradication of brucellosis. The present work aimed to determine of the most common brucella species biotypes infecting human and animals by using serological and multiplex PCR . A total of 170 samples (90 serum samples from male and 80 serum samples from female as well as the same number from male and female whole blood samples with ( EDTA anticoagulant) at different age and 190 animals sera and the same number of samples whole blood with EDTA anticoagulant collected from (135 cow & 55 buffaloes ) through the period between (February 2011 to March 2013 ) in Egypt . Their sera were tested by ELISA and SAT. DNA was extracted and examined by multiplex PCR ( MPCR) involving specific primers for Brucella species (B.SPP), Brucella melitensis (B. melitensis) and Brucella abortus (B. abortus) based on IS711 in the brucella chromosome. Brucellosis was confirmed in patient by ELISA IgM was ( 23.3%) in male and ( 17.5%) in female, brucella IgG in male was (37.7%) and in female was (26.25%) and by SAT was (25.5%) in male and (22.5%) in female at El Fayoum Governorate respectively. Comparing serological tests with MPCR in human (male and female ) serum and blood samples gave total positive of ELISA IgM (20.58 %) , ELISA IgG, (32.32 ), SAT ( 24.11% ) and MPCR (32.9 %). Also ELISA (18.5 % ), SAT (14.8) and MPCR (21.48% ) in cow and ELISA (14.5% ), SAT (12.7%) and MPCR (5.5%) in buffaloes in the same region. The present study proved that multiplex PCR assay is a rapid , sensitive and accurate technique for diagnosis of B.SPP biotype compared to ELSA IgM, IgG and SAT. However ELISA was simple, accurate, rapid, does not require specialized training or equipment and economical for the detection of Brucella antibody...
Routine serological diagnosis of toxoplasmosis provides high sensitivity, but specificity varies depending on the test used; false-positive results (IgM) have been reported. Blood samples were collected from 88 women (59 pregnant and 29 nonpregnant) and 86 contact animals (62 sheep and 24 goats) at El Fayoum Governorate during the period from October 2005 to December 2006. All collected samples were tested for Toxoplasma gondii infection by serological tests (ELISA IgM & IgG and Sabin-Feldman dye test) and polymerase chain reaction (PCR). Results revealed specific IgG in 45.8% and 41.4%, IgM in 30.5% and 24.2%, and positive Sabin-Feldman dye test in 23.7% and 17.2% in pregnant and nonpregnant women, respectively. Positive PCR products were detected in 32.2% and 27.6% in pregnant and nonpregnant women, respectively. Regarding animals, positive ELISA IgG and PCR were detected in 98.4% and 67.7% of sheep and 41.7% and 25.0% of goats, respectively. It was concluded that serological tests can detect higher rate of toxoplasmosis than PCR, so ELISA combined with the PCR technique is a recommended tool for accurate diagnosis of toxoplasmosis.
