Repository of Research and Investigative Information

Repository of Research and Investigative Information

Dezful University of Medical Sciences

In silico design and in vitro development of a highly accurate test to detect Brucella species

(2019) In silico design and in vitro development of a highly accurate test to detect Brucella species. Turkish Journal of Medical Sciences. pp. 368-374. ISSN 1300-0144

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Background: Conventional methods of detecting Brucella spp. suffer from technical and biological complications. Besides, newly characterized species of the genus Brucella could be neglected by previously designed polymerase chain reaction (PCR) tests. Therefore, a snore accurate PCR-based test seems to be imminently needed. Materials and methods: Blood samples were collected from 39 patients diagnosed with brucellosis and 25 healthy controls. Multiple sequence alignments (MSA) were performed on 500 Omp2-related protein and gene sequences. Thereafter, specific pruners were designed and synthesized for the regions with highest conservancy. The collected samples were assessed by PCR test. overcome the cross-reactivity issue, PCR thermal program was optimized regarding annealing time and temperature. Results: The MSA results indicated that the N terminus region of the Omp2 protein (DNA 5' end) is associated with highest conservancy. Primers with highest specificity were designed and synthesized. A two-step PCR reaction was successfully designed and optimized. The desirable bands were observed in clinical samples with high accuracy. Conclusion: It should be pointed out that using a precisely designed primer pair would bring about early infection detection, more success to detect all natural variants and higher cost-to-efficacy ratio in comparison to other detection methods.

Item Type: Article
Keywords: Brucellosis diagnosis Omp2 primer design multiple sequence alignment PCR pcr identification DNA sequences locus milk General & Internal Medicine
Page Range: pp. 368-374
Journal or Publication Title: Turkish Journal of Medical Sciences
Journal Index: ISI
Volume: 49
Number: 1
Identification Number:
ISSN: 1300-0144
Depositing User: مهندس مهدی شریفی

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