Biochemistry & Molecular Biology الكيمياء الحيوية والأحياء الجزيئية

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Browsing Biochemistry & Molecular Biology الكيمياء الحيوية والأحياء الجزيئية by Author "Ahmad Msallam Ibrahim Makhamreh"

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    Development of loop mediated isothermal DNA amplification (LAMP) test for the detection of active brucellosis infections among diary animals
    (Al-Quds University, 2021-05-29) Ahmad Msallam Ibrahim Makhamreh; أحمد مسلم إبراهيم مخامره
    Brucellosis is zoonotic a disease caused by bacterial genus Brucella. It affects a wide range of mammalian species and is transmissible to humans, making it a major socioeconomic problem. Because of the unspecific symptoms and signs that are shared with other febrile diseases, diagnosing brucellosis can be challenging and due to the slow growth rate in blood culture. Rose Bengal test is an internationally recommended test for the screening of brucellosis. They are not always specific as the antibodies can cross-react with other gram-negative bacteria, these tests are not of high value in detecting infection at early stages, May give a false negative result, for a short period after infection, Low sensitivity particularly in long chronic cases, relatively low specificity in endemic areas, since generation and disappearance of antibodies requires some time. To diagnose brucellosis, we developed a LAMP assay. The reaction takes 60 minutes to complete at 63 ° C. Its highly sensitive method , simple, quick, specific, and cost-effective. The strand displacement reaction is carried out at a constant temperature and is characterized by the use of four different primers, each of which is designed to recognize six distinct regions on the target gene. There was no cross-reactivity with the other bacteria. Its More effective than traditional PCR (Less expensive, easier, faster, no hardware required, and with the same accuracy as PCR). The blood test was carried out for 5000 sheep. out of which 200 (4%) were positive. We used the positive results (200) samples and re-examination using the LAMP method that we have developed. Whereas, by developing the LAMP method, we were able to eliminate the interference with other bacteria, and get rid of the false positive result due to the presence of antibodies to Brucella in the blood as a result of a previous infection. Whereas, The LAMP method, unlike the serology approach, is based on the detection of DNA for bacteria rather than antibodies to the bacterium, In addition to removing all of the barriers to detecting brucellosis that existed using serological approaches.