Issue 2

Journal for Veterinary Medicine, Biotechnology and Biosafety

Volume 4, Issue 2, June 2018, Page 43

ISSN 2411-3174 (print version) ISSN 2411-0388 (online version)

MOLECULAR TECHNIQUES FOR DIAGNOSIS AND CONTROL OF HUMAN AND ANIMAL BRUCELLA ISOLATES FROM GEORGIA

Sidamonidze K. 1, Ramishvili M. 1, Malania L. 1, Zhgenti E. 1, Donduashvili M. 2, Nikolaishvili M. 2, Vepkhvadze N. 2, Kokhreidze M. 2, Avaliani L. 3, Imnadze P. 1

1 National Center for Disease Control and Public Health, Tbilisi, Georgia, e-mail: m.ramishvili@ncdc.ge, pimnadze@ncdc.ge

2 Laboratory of the Ministry of Agriculture, Tbilisi, Georgia

3 National Food Agency, Tbilisi, Georgia

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Citation for print version: Sidamonidze, K., Ramishvili, M., Malania, L., Zhgenti, E., Donduashvili, M., Nikolaishvili, M., Vepkhvadze, N., Kokhreidze, M., Avaliani, L. and Imnadze, P. (2018) ‘Molecular techniques for diagnosis and control of human and animal Brucella isolates from Georgia’, Journal for Veterinary Medicine, Biotechnology and Biosafety, 4(2), p. 43.

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Citation for online version: Sidamonidze, K., Ramishvili, M., Malania, L., Zhgenti, E., Donduashvili, M., Nikolaishvili, M., Vepkhvadze, N., Kokhreidze, M., Avaliani, L. and Imnadze, P. (2018) ‘Molecular techniques for diagnosis and control of human and animal Brucella isolates from Georgia’, Journal for Veterinary Medicine, Biotechnology and Biosafety. [Online] 4(2), p. 43. Available at: http://jvmbbs.kharkov.ua/archive/2018/volume4/issue2/oJVMBBS_2018042_043.pdf

Brucellosis is an ancient and highly contagious zoonotic disease caused by the genus Brucella that remains to be a major public and animal health problem in Georgia and is considered one of the most important health disorders spread worldwide. The leading institutions conducting surveillance on brucellosis in Georgia are the National Center for Disease Control and Public Health under the Ministry of Health. the Laboratory of the Ministry of Agriculture, and the National Food Agency under the Ministry of Agriculture.

The aim of this study was evaluation of implemented molecular diagnostic tests for the assessment of animal and human cases of brucellosis in Georgia.

Isolates from human blood and ruminant milk or blood suspected as Brucella by microbiological, biochemical and serological testing were confirmed by real-time PCR (Brucella T1, Biofire Technology). Species identity of Brucella cultures were confirmed and typed by conventional AMOS PCR, Single Nucleotide Polymorphism (SNP) assays and later ‘Bruce-ladder’ assay. A multi-locus variable number tandem repeat analysis (MLVA) approach was applied for providing valuable information for epidemiological investigations.

AMOS PCR supported biochemical test results for 72 Brucella melitensis and four Brucella abortus strains, but not for 39 suspected B. abortus human and animal isolates. SNP typing of all 115 isolates supported the AMOS PCR results, but also confirmed suspected B. abortus species of the 39 strains not amplified by AMOS PCR. Above-mentioned Brucella strains were confirmed later also by ‘Bruce-ladder’ assay. In the present study, for the first time we have studied the genetic variability of 115 strains obtained in Georgia. Genotypes were revealed by a MLVA‑15 approach with good subspecies discriminatory capabilities providing valuable information for epidemiological investigations and obtained data were utilized for construction of the phylogenic tree using Bionumerics Software version 6.1.

Evidences of the molecular-genetic research first have confirmed the existence of B. melitensis and B. abortus strains circulating in humans and animals in Georgia. Basing on our results of molecular-genetic researches, we can suppose, that B. abortus 3, 5, 6 and/or 9 biovars are more frequently spread, than B. abortus 1, 2 and 4 in Georgia. Thus, we can say, that application of AMOS PCR method is limited in our country. Species-specific SNP typing and ‘Bruce-ladder’ assays resolved the difficulties caused by limitations of AMOS PCR to recognize all biovars of B. abortus. Obtained results suggest that diversity of Brucella strains in Georgia is greater than captured in this study and it needs continuation of a large-scale molecular-biological researches in this direction. Establishment and application of MLVA genotyping will serve invaluably to track the source of infection in case of bioterrorism or outbreak in Georgia or in surrounding areas. Thus, implementation of sustainable set of assays and a ‘One Health’ approach resulted in a more effective monitoring system for both human and animal brucellosis in Georgia.

Keywords: animal brucellosis, B. melitensis, B. abortus.