Detection Of Brucellosis In Sheep And Goats By Serum Agglutination Test (Sat) And Polymerase Chain Reaction (Pcr)
By: Dr. Imran Zafar
| Dr. M. Younus Rana.
Contributor(s): Dr. Matu-ur-Rehman Khan.
Material type: 
BookPublisher: 2010Subject(s): Department of PathologyDDC classification: 1190,T Dissertation note: In the current research project, a Polymerase chain reaction (PCR) was optimized by using omp 31 (outer membrane protein) DNA amplification and Serum agglutination test (SAT) was also evaluated for the species specific diagnosis of the brucellosis in sheep and goats.
Blood and serum samples from two hundred and fifty sheep and goats each were collected aseptically at different sheep and goat farms of Punjab Pakistan. Before performing Serum agglutination test (SAT), Rose Bengal Plate Test (RBPT) was performed as a screening test. RBPT screening of serum revealed nine (3.6%) positive samples out of two hundred and fifty sheep and goats. Out of nine positive samples, five (55.55 %) goats and four (44.44 %) sheep were positive. RBPT positive samples were further carried to Serum agglutination test (SAT). Out of nine positive samples, six (66.66 %) remained positive when SAT was applied but other three (33.33 %) samples gave the dilution below 1:40 which is considered to be negative (Alton et., al 1988). Polymerase chain reaction (PCR) was carried out on the blood samples. Genomic DNA was extracted by using Genomic DNA Purification Kit (Vivantis). The PCR was performed in a 50 µl reaction mixture. The tubes containing PCR mix were subjected to amplification cycles in a thermocycler after adjusting the amplification conditions. After completion of the amplification cycles, the PCR product was characterized by 1.2 % agarose gel electrophoresis along with 100 bp DNA ladder to estimate the size of the PCR product and the gel was photographed with a Polaroid camera. PCR gave eight (3.2%) positive results.
It was concluded from current study that polymerase chain reaction is a superior and sensitive test as compared to Serum agglutination test (SAT). The test is comparatively sensitive and can detect the brucella in cases of low bacteremia when they don't produce enough antibodies to be detected by other serological tests. The results of the study confirm that a polymerase chain reaction for brucellosis can provide with a sensitive and appropriate diagnostic tool.
| Item type | Current location | Collection | Call number | Status | Date due | Barcode | Item holds |
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Thesis
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UVAS Library Thesis Section | Veterinary Science | 1190,T (Browse shelf) | Available | 1190,T |
In the current research project, a Polymerase chain reaction (PCR) was optimized by using omp 31 (outer membrane protein) DNA amplification and Serum agglutination test (SAT) was also evaluated for the species specific diagnosis of the brucellosis in sheep and goats.
Blood and serum samples from two hundred and fifty sheep and goats each were collected aseptically at different sheep and goat farms of Punjab Pakistan. Before performing Serum agglutination test (SAT), Rose Bengal Plate Test (RBPT) was performed as a screening test. RBPT screening of serum revealed nine (3.6%) positive samples out of two hundred and fifty sheep and goats. Out of nine positive samples, five (55.55 %) goats and four (44.44 %) sheep were positive. RBPT positive samples were further carried to Serum agglutination test (SAT). Out of nine positive samples, six (66.66 %) remained positive when SAT was applied but other three (33.33 %) samples gave the dilution below 1:40 which is considered to be negative (Alton et., al 1988). Polymerase chain reaction (PCR) was carried out on the blood samples. Genomic DNA was extracted by using Genomic DNA Purification Kit (Vivantis). The PCR was performed in a 50 µl reaction mixture. The tubes containing PCR mix were subjected to amplification cycles in a thermocycler after adjusting the amplification conditions. After completion of the amplification cycles, the PCR product was characterized by 1.2 % agarose gel electrophoresis along with 100 bp DNA ladder to estimate the size of the PCR product and the gel was photographed with a Polaroid camera. PCR gave eight (3.2%) positive results.
It was concluded from current study that polymerase chain reaction is a superior and sensitive test as compared to Serum agglutination test (SAT). The test is comparatively sensitive and can detect the brucella in cases of low bacteremia when they don't produce enough antibodies to be detected by other serological tests. The results of the study confirm that a polymerase chain reaction for brucellosis can provide with a sensitive and appropriate diagnostic tool.
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