Reliability of molecular tests in diagnosing ovine brucellosis caused by Brucella ovis
Authors
Zelenović, MladenMarinković, Darko
Stević, Nataša
Stanojević, Slavoljub
Aničić, Milan
Milićević, Vesna
Valčić, Olivera
Radojičić, Sonja
Article (Published version)
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Brucella ovis infects sheep and causes a clinical or subclinical disease characterized by genital lesions and reduced fertility in rams, placentitis and abortions in ewes, and increased perinatal mortality in lambs. As part of this study, analyses were conducted on a sheep farm near Belgrade (Serbia). Of the serological tests, indirect ELISA was performed. A total of 94 blood sera were analyzed, 33 from rams and 61 from ewes. The results showed 23 (69.7%) positive fi ndings in rams and 2 (3.3%) positive fi ndings in ewes, with an overall prevalence of 25.4% bounded by a 95% confi dence interval. Bruce-ladder multiplex PCR and Real time PCR were performed on 19 serologically positive rams and two serologically positive ewes. The results indicate a slightly higher sensitivity of Real time PCR compared to conventional PCR in diagnosing B. ovis from the reproductive tissues of rams. However, the differences in sensitivity between different nucleic acid extraction protocols were not signifi... cant. Most rams showed a positive PCR result in only one sample of reproductive tissue, suggesting the recommendation to take multiple samples from each animal. Further research is needed to bring the sensitivity of molecular tests in diagnosing ram epididymitis closer to the sensitivity of serological tests.
Brucella ovis infi cira ovce i uzrokuje kliničku ili subkliničku bolest koja se karakteriše genitalnim lezijama i smanjenom plodnošću kod ovnova, placentitisom i pobačajima kod ovaca te povećanom neonatalnom smrtnošću kod jaganjaca. U okviru ovog istraživanja, sprovedene su analize na farmi ovaca u blizini Beograda (Srbija). Od seroloških testova, rađena je indirektna ELISA. Ukupno je analizirano 94 krvna seruma, 33 od ovnova i 61 od ovaca. Rezultati su pokazali 23 (69,7%) pozitivna nalaza kod ovnova i 2 (3,3%) pozitivna nalaza kod ovaca, s ukupnom prevalencijom od 25,4% uz 95% interval pouzdanosti. Bruce-ladder multiplex PCR i Real time PCR su rađeni kod 19 serološki pozitivnih ovnova i dve serološki pozitivne ovce. Rezultati pokazuju da je Real time PCR pokazao nešto veću osetljivost u poređenju s konvencionalnim PCR-om u dijagnozi B. ovis iz reproduktivnih tkiva ovnova. Ipak, razlike u osetljivosti između različitih protokola ekstrakcije nukleinske kiseline nisu bile značajne. Većin...a ovnova je imala pozitivan PCR rezultat samo kod jednog zbirnog uzorka reproduktivnog tkiva, što ukazuje na preporuku uzimanja više uzoraka od svake životinje. Potrebna su dalja istraživanja kako bi se osetljivost molekularnih testova u dijagnozi epididimitisa ovnova približila osetljivosti seroloških testova.
Keywords:
Brucella ovis / ELISA / molecular diagnostics / sheepSource:
Acta Veterinaria-Beograd, 2024, 74, 1, 133-144Publisher:
- Belgrade : Faculty of Veterinary Medicine, University of Belgrade
Funding / projects:
- Ministry of Science, Technological Development and Innovation of the Republic of Serbia, institutional funding - 200143 (University of Belgrade, Faculty of Veterinary Medicine) (RS-MESTD-inst-2020-200143)
DOI: 10.2478/acve-2024-0010
ISSN: 0567-8315
WoS: 001185980700010
Scopus: 2-s2.0-85187982842
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Naučni institut za veterinarstvo SrbijeTY - JOUR AU - Zelenović, Mladen AU - Marinković, Darko AU - Stević, Nataša AU - Stanojević, Slavoljub AU - Aničić, Milan AU - Milićević, Vesna AU - Valčić, Olivera AU - Radojičić, Sonja PY - 2024 UR - https://reponivs.nivs.rs/handle/123456789/956 AB - Brucella ovis infects sheep and causes a clinical or subclinical disease characterized by genital lesions and reduced fertility in rams, placentitis and abortions in ewes, and increased perinatal mortality in lambs. As part of this study, analyses were conducted on a sheep farm near Belgrade (Serbia). Of the serological tests, indirect ELISA was performed. A total of 94 blood sera were analyzed, 33 from rams and 61 from ewes. The results showed 23 (69.7%) positive fi ndings in rams and 2 (3.3%) positive fi ndings in ewes, with an overall prevalence of 25.4% bounded by a 95% confi dence interval. Bruce-ladder multiplex PCR and Real time PCR were performed on 19 serologically positive rams and two serologically positive ewes. The results indicate a slightly higher sensitivity of Real time PCR compared to conventional PCR in diagnosing B. ovis from the reproductive tissues of rams. However, the differences in sensitivity between different nucleic acid extraction protocols were not signifi cant. Most rams showed a positive PCR result in only one sample of reproductive tissue, suggesting the recommendation to take multiple samples from each animal. Further research is needed to bring the sensitivity of molecular tests in diagnosing ram epididymitis closer to the sensitivity of serological tests. AB - Brucella ovis infi cira ovce i uzrokuje kliničku ili subkliničku bolest koja se karakteriše genitalnim lezijama i smanjenom plodnošću kod ovnova, placentitisom i pobačajima kod ovaca te povećanom neonatalnom smrtnošću kod jaganjaca. U okviru ovog istraživanja, sprovedene su analize na farmi ovaca u blizini Beograda (Srbija). Od seroloških testova, rađena je indirektna ELISA. Ukupno je analizirano 94 krvna seruma, 33 od ovnova i 61 od ovaca. Rezultati su pokazali 23 (69,7%) pozitivna nalaza kod ovnova i 2 (3,3%) pozitivna nalaza kod ovaca, s ukupnom prevalencijom od 25,4% uz 95% interval pouzdanosti. Bruce-ladder multiplex PCR i Real time PCR su rađeni kod 19 serološki pozitivnih ovnova i dve serološki pozitivne ovce. Rezultati pokazuju da je Real time PCR pokazao nešto veću osetljivost u poređenju s konvencionalnim PCR-om u dijagnozi B. ovis iz reproduktivnih tkiva ovnova. Ipak, razlike u osetljivosti između različitih protokola ekstrakcije nukleinske kiseline nisu bile značajne. Većina ovnova je imala pozitivan PCR rezultat samo kod jednog zbirnog uzorka reproduktivnog tkiva, što ukazuje na preporuku uzimanja više uzoraka od svake životinje. Potrebna su dalja istraživanja kako bi se osetljivost molekularnih testova u dijagnozi epididimitisa ovnova približila osetljivosti seroloških testova. PB - Belgrade : Faculty of Veterinary Medicine, University of Belgrade T2 - Acta Veterinaria-Beograd T1 - Reliability of molecular tests in diagnosing ovine brucellosis caused by Brucella ovis EP - 144 IS - 1 SP - 133 VL - 74 DO - 10.2478/acve-2024-0010 ER -
@article{ author = "Zelenović, Mladen and Marinković, Darko and Stević, Nataša and Stanojević, Slavoljub and Aničić, Milan and Milićević, Vesna and Valčić, Olivera and Radojičić, Sonja", year = "2024", abstract = "Brucella ovis infects sheep and causes a clinical or subclinical disease characterized by genital lesions and reduced fertility in rams, placentitis and abortions in ewes, and increased perinatal mortality in lambs. As part of this study, analyses were conducted on a sheep farm near Belgrade (Serbia). Of the serological tests, indirect ELISA was performed. A total of 94 blood sera were analyzed, 33 from rams and 61 from ewes. The results showed 23 (69.7%) positive fi ndings in rams and 2 (3.3%) positive fi ndings in ewes, with an overall prevalence of 25.4% bounded by a 95% confi dence interval. Bruce-ladder multiplex PCR and Real time PCR were performed on 19 serologically positive rams and two serologically positive ewes. The results indicate a slightly higher sensitivity of Real time PCR compared to conventional PCR in diagnosing B. ovis from the reproductive tissues of rams. However, the differences in sensitivity between different nucleic acid extraction protocols were not signifi cant. Most rams showed a positive PCR result in only one sample of reproductive tissue, suggesting the recommendation to take multiple samples from each animal. Further research is needed to bring the sensitivity of molecular tests in diagnosing ram epididymitis closer to the sensitivity of serological tests., Brucella ovis infi cira ovce i uzrokuje kliničku ili subkliničku bolest koja se karakteriše genitalnim lezijama i smanjenom plodnošću kod ovnova, placentitisom i pobačajima kod ovaca te povećanom neonatalnom smrtnošću kod jaganjaca. U okviru ovog istraživanja, sprovedene su analize na farmi ovaca u blizini Beograda (Srbija). Od seroloških testova, rađena je indirektna ELISA. Ukupno je analizirano 94 krvna seruma, 33 od ovnova i 61 od ovaca. Rezultati su pokazali 23 (69,7%) pozitivna nalaza kod ovnova i 2 (3,3%) pozitivna nalaza kod ovaca, s ukupnom prevalencijom od 25,4% uz 95% interval pouzdanosti. Bruce-ladder multiplex PCR i Real time PCR su rađeni kod 19 serološki pozitivnih ovnova i dve serološki pozitivne ovce. Rezultati pokazuju da je Real time PCR pokazao nešto veću osetljivost u poređenju s konvencionalnim PCR-om u dijagnozi B. ovis iz reproduktivnih tkiva ovnova. Ipak, razlike u osetljivosti između različitih protokola ekstrakcije nukleinske kiseline nisu bile značajne. Većina ovnova je imala pozitivan PCR rezultat samo kod jednog zbirnog uzorka reproduktivnog tkiva, što ukazuje na preporuku uzimanja više uzoraka od svake životinje. Potrebna su dalja istraživanja kako bi se osetljivost molekularnih testova u dijagnozi epididimitisa ovnova približila osetljivosti seroloških testova.", publisher = "Belgrade : Faculty of Veterinary Medicine, University of Belgrade", journal = "Acta Veterinaria-Beograd", title = "Reliability of molecular tests in diagnosing ovine brucellosis caused by Brucella ovis", pages = "144-133", number = "1", volume = "74", doi = "10.2478/acve-2024-0010" }
Zelenović, M., Marinković, D., Stević, N., Stanojević, S., Aničić, M., Milićević, V., Valčić, O.,& Radojičić, S.. (2024). Reliability of molecular tests in diagnosing ovine brucellosis caused by Brucella ovis. in Acta Veterinaria-Beograd Belgrade : Faculty of Veterinary Medicine, University of Belgrade., 74(1), 133-144. https://doi.org/10.2478/acve-2024-0010
Zelenović M, Marinković D, Stević N, Stanojević S, Aničić M, Milićević V, Valčić O, Radojičić S. Reliability of molecular tests in diagnosing ovine brucellosis caused by Brucella ovis. in Acta Veterinaria-Beograd. 2024;74(1):133-144. doi:10.2478/acve-2024-0010 .
Zelenović, Mladen, Marinković, Darko, Stević, Nataša, Stanojević, Slavoljub, Aničić, Milan, Milićević, Vesna, Valčić, Olivera, Radojičić, Sonja, "Reliability of molecular tests in diagnosing ovine brucellosis caused by Brucella ovis" in Acta Veterinaria-Beograd, 74, no. 1 (2024):133-144, https://doi.org/10.2478/acve-2024-0010 . .