Instituto Nacional de Investigação Agrária e Veterinária- IP, Strategic Unit of Animal Health-. Laboratory of Bacteriology and Mycology, Lisbon, Portugal. 3.
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5TH CONGRESS OF THE EUROPEAN ASSOCIATION OF VETERINARY LABORATORY DIAGNOSTICIANS 14 – 17 OCTOBER, 2018, MCE BUSINESS AND CONFERENCE CENTRE, BRUSSELS, BELGIUM
OP07 - O N E H E A LTH : FO O D B O R N E P A T H OGEN S, Z OON OSI S A N D A N T I B I OR ESI ST A N C E - OR A L PRESEN TA TIO N S
O 26 FIRST LINE TYPING FOR BRUCELLA ABORTUS AND BRUCELLA MELITENSIS G. Garofolo1, A. Janowicz1, K. Zilli1, A.C. Ferreira2, A.S. Simões2, V. Blanda3, S. Villari3, M. Babetsa4, E. Boukouvala4, G. Wareth5, M. El-Diasty6, H. Neubauer5, M. Tittarelli1, F. Sacchini1, F. De Massis1, L. Sacchini1 1
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Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise “G. Caporale”, National and OIE Reference Laboratory for Brucellosis, Teramo, Italy Instituto Nacional de Investigação Agrária e Veterinária- IP, Strategic Unit of Animal HealthLaboratory of Bacteriology and Mycology, Lisbon, Portugal Istituto Zooprofilattico Sperimentale della Sicilia “A.Mirri”, Entomology and control of environmental vectors, Palermo, Italy Hellenic Agricultural Organisation- DEMETER- Veterinary Research Institute, Laboratory of molecular biology/microbiology, Thessaloníki, Greece Friedrich-Loeffler-Institut- Institute of Bacterial Infections and Zoonoses, National/OIE Reference Laboratory for Brucellosis, Jena, Germany Animal Health Research Institute-Mansoura Provincial Laboratory, Department of brucellosis, Mansoura, Egypt
Introduction: Brucella abortus and B. melitensis are genetically monomorphic and, therefore, an accurate comparison of isolates within one species using molecular typing can often be challenging. Nonetheless, MLVA is considered a reliable tool for identification and investigation of brucellosis outbreaks. To date, 9 different MLVA schemes utilizing 34 VNTRs have been proposed. Our aim was to design new MLVA panels in order to simplify the typing strategy without compromising accuracy. Materials and Methods: A set of 202 Brucella isolates from Italy, Portugal, Egypt and Greece were typed using 34 VNTRs. To select the best typing markers, we calculated their discriminatory capacity, the linkage disequilibrium and the index of association. The performance and the convenience criteria were evaluated and compared with the previous methodologies. The data obtained were deposited in the MLVA web repository of the BrucMedNet project. Results: Based on the MLVA analysis, we selected two different panels dedicated to B. abortus and B. melitensis (Table 1).
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Each was composed of 7 VNTRs, which had 100 % of typeability, a good association index and a medium or high Simpson diversity index (SDI). These panels were in good concordance with the published panels and resulted in a similar or higher number of genotypes (Figure 1).
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5TH CONGRESS OF THE EUROPEAN ASSOCIATION OF VETERINARY LABORATORY DIAGNOSTICIANS 14 – 17 OCTOBER, 2018, MCE BUSINESS AND CONFERENCE CENTRE, BRUSSELS, BELGIUM
Discussion and Conclusion: Although WGS is revolutionizing the molecular epidemiology tools, MLVA is still an effective first line typing array due to its rapid turnaround time and the lower cost. The new schemes aim to replace the previous MLVA panels and will contribute to an efficient, fast and cost-effective routine tests for the surveillance of brucellosis.
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