Molecular typing of Brucella melitensis endemic strains and differentiation from the vaccine strain Rev-1

Georgios Noutsios, Rigini M. Papi, Loukia V. Ekateriniadou, Anastasios Minas, Dimitrios A. Kyriakidis

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

In the present study forty-four Greek endemic strains of Br. melitensis and three reference strains were genotyped by Multi locus Variable Number Tandem Repeat (ML-VNTR) analysis based on an eight-base pair tandem repeat sequence that was revealed in eight loci of Br. melitensis genome. The forty-four strains were discriminated from the vaccine strain Rev-1 by Restriction Fragment Length Polymorphism (RFLP) and Denaturant Gradient Gel Electrophoresis (DGGE). The ML-VNTR analysis revealed that endemic, reference and vaccine strains are genetically closely related, while most of the loci tested (1, 2, 4, 5 and 7) are highly polymorphic with Hunter-Gaston Genetic Diversity Index (HGDI) values in the range of 0.939 to 0.775. Analysis of ML-VNTRs loci stability through in vitro passages proved that loci 1 and 5 are non stable. Therefore, vaccine strain can be discriminated from endemic strains by allele's clusters of loci 2, 4, 6 and 7. RFLP and DGGE were also employed to analyse omp2 gene and reveled different patterns among Rev-1 and endemic strains. In RFLP, Rev-1 revealed three fragments (282, 238 and 44 bp), while endemic strains two fragments (238 and 44 bp). As for DGGE, the electrophoretic mobility of Rev-1 is different from the endemic strains due to heterologous binding of DNA chains of omp2a and omp2b gene. Overall, our data show clearly that it is feasible to genotype endemic strains of Br. melitensis and differentiate them from vaccine strain Rev-1 with ML-VNTR, RFLP and DGGE techniques. These tools can be used for conventional investigations in brucellosis outbreaks.

Original languageEnglish (US)
Pages (from-to)7-20
Number of pages14
JournalVeterinary Research Communications
Volume36
Issue number1
DOIs
StatePublished - Mar 1 2012
Externally publishedYes

Fingerprint

Brucella melitensis
Molecular Typing
Minisatellite Repeats
Restriction Fragment Length Polymorphisms
Electrophoresis
Vaccines
Gels
vaccines
minisatellite repeats
loci
denaturing gradient gel electrophoresis
Tandem Repeat Sequences
Brucellosis
restriction fragment length polymorphism
Base Pairing
Genes
Disease Outbreaks
Alleles
Genotype
Genome

Keywords

  • Brucella melitensis
  • DGGE
  • ML-VNTR analysis
  • omp2 polymorphism
  • Phylogenetic relationship
  • Rev-1

ASJC Scopus subject areas

  • veterinary(all)

Cite this

Molecular typing of Brucella melitensis endemic strains and differentiation from the vaccine strain Rev-1. / Noutsios, Georgios; Papi, Rigini M.; Ekateriniadou, Loukia V.; Minas, Anastasios; Kyriakidis, Dimitrios A.

In: Veterinary Research Communications, Vol. 36, No. 1, 01.03.2012, p. 7-20.

Research output: Contribution to journalArticle

Noutsios, Georgios ; Papi, Rigini M. ; Ekateriniadou, Loukia V. ; Minas, Anastasios ; Kyriakidis, Dimitrios A. / Molecular typing of Brucella melitensis endemic strains and differentiation from the vaccine strain Rev-1. In: Veterinary Research Communications. 2012 ; Vol. 36, No. 1. pp. 7-20.
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