Assessment of PCR-Based DNA Fingerprinting Techniques as a Novel Approach for Genotyping of Brucella Strains in Egypt
Assessment of PCR-Based DNA Fingerprinting Techniques as a Novel Approach for Genotyping of Brucella Strains in Egypt
Nour H. Abdel-Hamid1*, Walid Elmonir2, Eman I. M. Beleta1, Rania I. Ismail1, Momtaz Shahein1, Mahmoud E. R. Hamdy1
ABSTRACT
This study aimed to evaluate two PCR-based DNA fingerprinting techniques: Enterobacterial repetitive intergenic consensus (ERIC) and Random amplification polymorphism DNA (RAPD), for genotyping the Brucella isolates, as alternative rapid tools for epidemiological tracing and investigation of brucellosis in Egypt. We recovered Brucella isolates (n=29) from different host species and governorates. The isolates were identified by the bacteriological and molecular techniques (AMOS-PCR) as Brucella (B) melitensis biovar 3 (n=24) and B. abortus bv1 (n=5). The ERIC and RAPD primers (ERIC2 and Operon 18/ RAPD4) used in this study created polymorphic band patterns in all Brucella isolates and the reference strains. ERIC-PCR and RAPD-PCR Dendrograms clustered the B. melitensis isolates into two clusters and three clusters composed of 16 and 13 genotypes with genetic similarity percentages of 62% and 54% with a diversity index of 0.82 and 0.88. Both ERIC-PCR and RAPD-PCR dendrograms clustered B. abortus isolates into two clusters and three genotypes with discriminatory of 0.8 and 0.72. Both ERIC and RAPD PCRs revealed 25 singleton genotypes (G) out of 35 genotypes. DNA-based typing techniques showed substantial diversity among the Brucella isolates. The genetic diversity among Brucella genotypes obtained in this study may suggest that the Brucella strains have been introduced into the country either intermittently or over a point of time and location. Brucella melitensis transmission among non-preferable hosts and passages of Brucella strains are likely to have resulted in such heterogeneity. We concluded that the acceptable diversity induced by ERIC-PCR and RAPD PCR allows sufficient discrimination between Brucella isolates. These findings recommend the application of these techniques as an inexpensive and quick tool for traceability of B. melitensis and B. abortus strains, especially in developing countries with limited resources needed for other advanced sequence-based genotyping tools. Further studies on a large scale are required to ensure the reproducibility of both techniques.
Keywords | Brucella, dendrogram, ERIC-PCR, genotypes, RAPD-PCR.
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