Application of fluorescence based molecular assays for improved detection and typing of Brucella strains in clinical samples
Bacteria from the genus Brucella are causative agents of brucellosis – a zoonotic disease which affects many wild and domestic animal species and humans. Taking into account the significant socio-economic and public health impact of brucellosis, its control is of great importance for endemic areas....
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Ss.Cyril and Methodius University, Skopje
2015-10-01
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| Series: | Macedonian Veterinary Review |
| Subjects: | |
| Online Access: | http://macvetrev.mk/2015-2/MacVetRev2015-38-2-14.pdf |
| _version_ | 1811297378506899456 |
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| author | Kiril Krstevski Ivancho Naletoski Dine Mitrov Slavcho Mrenoshki Iskra Cvetkovikj Aleksandar Janevski Aleksandar Dodovski Igor Djadjovski |
| author_facet | Kiril Krstevski Ivancho Naletoski Dine Mitrov Slavcho Mrenoshki Iskra Cvetkovikj Aleksandar Janevski Aleksandar Dodovski Igor Djadjovski |
| author_sort | Kiril Krstevski |
| collection | DOAJ |
| description | Bacteria from the genus Brucella are causative agents of brucellosis – a zoonotic disease which affects many wild and domestic animal species and humans. Taking into account the significant socio-economic and public health impact of brucellosis, its control is of great importance for endemic areas. The chosen control strategy could be successful only if adapted to the current epidemiological situation. This implies that a choice of appropriate diagnostic procedures for detection and typing of Brucella spp. strains are of essential importance. Significant advancement of molecular techniques and their advantages compared to classical methods, give strong arguments in promotion of these techniques as a powerful tool for comprehensive diagnostics of brucellosis. Considering this, the major tasks of the study were to select and implement molecular tests for detection and genotyping Brucella spp. and evaluate their performances using DNA from cultivated brucellae (islolates) and limited number of tissue samples from seropositive animals. The obtained results confirmed that implemented real time PCR for Brucella spp. detection, as well as MLVA-16 used for genotyping, have excellent analytical sensitivity (4.2 fg of Brucella DNA were successfully detected and genotyped). Furthermore, compared to bacteriological cultivation of Brucella spp., real time PCR and MLVA-16 protocols showed superior diagnostic sensitivity and detected Brucella DNA in tissues from which Brucella could not be cultivated. Based on the summarized study results, we propose a diagnostic algorithm for detection and genotyping of Brucella spp. bacteria. Routine use of proposed diagnostic algorithm will improve the effectiveness of infection confirmation and help for accurate evaluation of epidemiological situation. |
| first_indexed | 2024-04-13T06:03:19Z |
| format | Article |
| id | doaj.art-8f5c20dc623e485ea6f0383f50c99d4d |
| institution | Directory Open Access Journal |
| issn | 1409-7621 1857-7415 |
| language | English |
| last_indexed | 2024-04-13T06:03:19Z |
| publishDate | 2015-10-01 |
| publisher | Ss.Cyril and Methodius University, Skopje |
| record_format | Article |
| series | Macedonian Veterinary Review |
| spelling | doaj.art-8f5c20dc623e485ea6f0383f50c99d4d2022-12-22T02:59:20ZengSs.Cyril and Methodius University, SkopjeMacedonian Veterinary Review1409-76211857-74152015-10-0138222323210.14432/j.macvetrev.2015.09.055Application of fluorescence based molecular assays for improved detection and typing of Brucella strains in clinical samplesKiril Krstevski 0Ivancho Naletoski1Dine Mitrov2Slavcho Mrenoshki3Iskra Cvetkovikj4Aleksandar Janevski5Aleksandar Dodovski6Igor Djadjovski7Veterinary Institute, Faculty of Veterinary Medicine, Ss. Cyril and Methodius University in Skopje, Republic of MacedoniaAnimal Production and Health Section, Joint FAO/IAEA Division, International Atomic Energy Agency, Vienna, AustriaVeterinary Institute, Faculty of Veterinary Medicine, Ss. Cyril and Methodius University in Skopje, Republic of MacedoniaVeterinary Institute, Faculty of Veterinary Medicine, Ss. Cyril and Methodius University in Skopje, Republic of MacedoniaVeterinary Institute, Faculty of Veterinary Medicine, Ss. Cyril and Methodius University in Skopje, Republic of MacedoniaVeterinary Institute, Faculty of Veterinary Medicine, Ss. Cyril and Methodius University in Skopje, Republic of MacedoniaVeterinary Institute, Faculty of Veterinary Medicine, Ss. Cyril and Methodius University in Skopje, Republic of MacedoniaVeterinary Institute, Faculty of Veterinary Medicine, Ss. Cyril and Methodius University in Skopje, Republic of MacedoniaBacteria from the genus Brucella are causative agents of brucellosis – a zoonotic disease which affects many wild and domestic animal species and humans. Taking into account the significant socio-economic and public health impact of brucellosis, its control is of great importance for endemic areas. The chosen control strategy could be successful only if adapted to the current epidemiological situation. This implies that a choice of appropriate diagnostic procedures for detection and typing of Brucella spp. strains are of essential importance. Significant advancement of molecular techniques and their advantages compared to classical methods, give strong arguments in promotion of these techniques as a powerful tool for comprehensive diagnostics of brucellosis. Considering this, the major tasks of the study were to select and implement molecular tests for detection and genotyping Brucella spp. and evaluate their performances using DNA from cultivated brucellae (islolates) and limited number of tissue samples from seropositive animals. The obtained results confirmed that implemented real time PCR for Brucella spp. detection, as well as MLVA-16 used for genotyping, have excellent analytical sensitivity (4.2 fg of Brucella DNA were successfully detected and genotyped). Furthermore, compared to bacteriological cultivation of Brucella spp., real time PCR and MLVA-16 protocols showed superior diagnostic sensitivity and detected Brucella DNA in tissues from which Brucella could not be cultivated. Based on the summarized study results, we propose a diagnostic algorithm for detection and genotyping of Brucella spp. bacteria. Routine use of proposed diagnostic algorithm will improve the effectiveness of infection confirmation and help for accurate evaluation of epidemiological situation.http://macvetrev.mk/2015-2/MacVetRev2015-38-2-14.pdfbrucellosisclinical samplesDNAreal time PCRMLVA-16 |
| spellingShingle | Kiril Krstevski Ivancho Naletoski Dine Mitrov Slavcho Mrenoshki Iskra Cvetkovikj Aleksandar Janevski Aleksandar Dodovski Igor Djadjovski Application of fluorescence based molecular assays for improved detection and typing of Brucella strains in clinical samples Macedonian Veterinary Review brucellosis clinical samples DNA real time PCR MLVA-16 |
| title | Application of fluorescence based molecular assays for improved detection and typing of Brucella strains in clinical samples |
| title_full | Application of fluorescence based molecular assays for improved detection and typing of Brucella strains in clinical samples |
| title_fullStr | Application of fluorescence based molecular assays for improved detection and typing of Brucella strains in clinical samples |
| title_full_unstemmed | Application of fluorescence based molecular assays for improved detection and typing of Brucella strains in clinical samples |
| title_short | Application of fluorescence based molecular assays for improved detection and typing of Brucella strains in clinical samples |
| title_sort | application of fluorescence based molecular assays for improved detection and typing of brucella strains in clinical samples |
| topic | brucellosis clinical samples DNA real time PCR MLVA-16 |
| url | http://macvetrev.mk/2015-2/MacVetRev2015-38-2-14.pdf |
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