التنميط الحيوي لجرثومة البروسيلا المعزولة من الأبقار باستخدام تقنية تفاعل سلسلة البلمرة نوع تقييد طول الجزء المتعدد الأشكال (RFLP-PCR)‎ في مدينة الديوانية

الملخص EN

The current study was aimed to characterize the Brucella abortus in cows at molecular level after isolation and identification morphologically and serology by using the amplification of specific primers of omp2a gene that encodes to proteins of outer membrane of Brucella by using a technique of Restriction Fragment Length Polymorphism – Polymerase Chain Reaction (RFLP- PCR) .

A Total of 105 samples were collected from cow blood that clinically suspected with brucellosis for the period of November /2011 to May / 2012 from animal shelters in different areas of AL-Diwaniya city .

Blood sera were prepared to conduct the Laboratory tests which included the serological tests (Rose Bengal (RBT) and Tube Agglutination (TAT) ) ,then the Brucella was isolated on selective media such as Brucella agar media which confirmed molecularly by using classical Polymerase Chain Reaction to amplify the primers of omp2a gene and finlly the technique of RFLP- PCR was used to determine the species and biovars of Brucella after using the restriction endonuclease (PSTI) for Amplification products of omp2a gene .

the results of Brucella isolation On Brucella agar was 15/18(83.3%), Results of classical Polymerase Chain Reaction (PCR) as confirming test for Brucella isolates after extraction of DNA and amplification of primers belong to omp2a gene showed the appearance of single band on agarose gel represented the DNA amplified (amplicon ) with molecular size (1100 bp) , and the percent of Brucella detection by this technique was 13/15(12.3%) .The results of (RFLP-PCR) for differention among species and biovars of Brucella revealed the appearance of two distinct band on agarose gel that had a molecular sizes (200 and 550 bp) which belong to B.melitensis biovars 1,3 and B.abortus biovars 3,5,6,9 , respectively.