TY  - JOUR
AU  - Milovanović, Milovan
AU  - Milićević, Vesna
AU  - Radojičić, Sonja
AU  - Valčić, Miroslav
AU  - Hoffmann, Bernd
AU  - Dietze, Klaas
PY  - 2020
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/1795
AB  - Background The detection of antibodies against capripoxvirus has become easier with a commercially available ELISA validated for serum and plasma. In order to explore its suitability for immunological investigations on alternative samples, this study targeted milk as sample matrix available through non-invasive sampling. Methods Samples for this study were collected from dairy cows vaccinated against LSD in an area without reported LSD virus circulation. Paired serum and milk (individual and bulk) samples were tested by ELISA without and with modifications of the sample incubation time for the milk samples. For the evaluation of the test specificity, 352 milk samples from a milk repository in Germany were used as negative control. Receiver operating characteristic analysis was performed for determination of the Youden index and determination of the most suitable cut-off value for maximum specificity. Results From 154 analyzed serum samples from Serbia, 75 were detected as positive in the ELISA. Sensitivity and specificity of the ELISA test for milk samples reached values of 88 to 91% using Youden criteria. A cut-off of 10 was determined aiming for maximum specificity. This cut-off value was used for further analysis. Using the protocol for serum, out of 154 milk samples, 38 were detected as positive, number of positive detected milk samples increase up to 48 with modified protocol. Milk samples from Germany reacted negative, except two samples that had borderline results using modified protocol. Significant statistical difference (p < 0.05) was observed between two incubation protocols. The detection of LSD-specific antibodies from bulk milk samples (pools of 2-10 individuals) came along with a reduced sensitivity over the sample of individual animals. Conclusions Results show that the detection of capripoxvirus specific antibodies in milk samples using the commercially available ELISA from IDvet is feasible and can represent a helpful tool for LSDV monitoring programs.
PB  - BMC, London
T2  - Virology Journal
T1  - Suitability of individual and bulk milk samples to investigate the humoral immune response to lumpy skin disease vaccination by ELISA
VL  - 17
IS  - 1
SP  - 28
DO  - 10.1186/s12985-020-01298-x
ER  - 

@article{
author = "Milovanović, Milovan and Milićević, Vesna and Radojičić, Sonja and Valčić, Miroslav and Hoffmann, Bernd and Dietze, Klaas",
year = "2020",
abstract = "Background The detection of antibodies against capripoxvirus has become easier with a commercially available ELISA validated for serum and plasma. In order to explore its suitability for immunological investigations on alternative samples, this study targeted milk as sample matrix available through non-invasive sampling. Methods Samples for this study were collected from dairy cows vaccinated against LSD in an area without reported LSD virus circulation. Paired serum and milk (individual and bulk) samples were tested by ELISA without and with modifications of the sample incubation time for the milk samples. For the evaluation of the test specificity, 352 milk samples from a milk repository in Germany were used as negative control. Receiver operating characteristic analysis was performed for determination of the Youden index and determination of the most suitable cut-off value for maximum specificity. Results From 154 analyzed serum samples from Serbia, 75 were detected as positive in the ELISA. Sensitivity and specificity of the ELISA test for milk samples reached values of 88 to 91% using Youden criteria. A cut-off of 10 was determined aiming for maximum specificity. This cut-off value was used for further analysis. Using the protocol for serum, out of 154 milk samples, 38 were detected as positive, number of positive detected milk samples increase up to 48 with modified protocol. Milk samples from Germany reacted negative, except two samples that had borderline results using modified protocol. Significant statistical difference (p < 0.05) was observed between two incubation protocols. The detection of LSD-specific antibodies from bulk milk samples (pools of 2-10 individuals) came along with a reduced sensitivity over the sample of individual animals. Conclusions Results show that the detection of capripoxvirus specific antibodies in milk samples using the commercially available ELISA from IDvet is feasible and can represent a helpful tool for LSDV monitoring programs.",
publisher = "BMC, London",
journal = "Virology Journal",
title = "Suitability of individual and bulk milk samples to investigate the humoral immune response to lumpy skin disease vaccination by ELISA",
volume = "17",
number = "1",
pages = "28",
doi = "10.1186/s12985-020-01298-x"
}

Milovanović, M., Milićević, V., Radojičić, S., Valčić, M., Hoffmann, B.,& Dietze, K.. (2020). Suitability of individual and bulk milk samples to investigate the humoral immune response to lumpy skin disease vaccination by ELISA. in Virology Journal
BMC, London., 17(1), 28.
https://doi.org/10.1186/s12985-020-01298-x

Milovanović M, Milićević V, Radojičić S, Valčić M, Hoffmann B, Dietze K. Suitability of individual and bulk milk samples to investigate the humoral immune response to lumpy skin disease vaccination by ELISA. in Virology Journal. 2020;17(1):28.
doi:10.1186/s12985-020-01298-x .

Milovanović, Milovan, Milićević, Vesna, Radojičić, Sonja, Valčić, Miroslav, Hoffmann, Bernd, Dietze, Klaas, "Suitability of individual and bulk milk samples to investigate the humoral immune response to lumpy skin disease vaccination by ELISA" in Virology Journal, 17, no. 1 (2020):28,
https://doi.org/10.1186/s12985-020-01298-x . .

TY  - JOUR
AU  - Milovanović, Milovan
AU  - Dietze, Klaas
AU  - Milićević, Vesna
AU  - Radojičić, Sonja
AU  - Valčić, Miroslav
AU  - Moritz, Tom
AU  - Hoffmann, Bernd
PY  - 2019
UR  - https://vet-erinar.vet.bg.ac.rs/handle/123456789/1732
AB  - BackgroundIn the presented study we investigated the development of the humoral immune response against LSDV during the process of re-vaccination of cattle over a time span of 5 months. In addition, the performance of different serological techniques for antibody detection against LSDV was compared. For sample collection, an area without previous LSD outbreak reports in Serbia was selected. Seventy-nine cattle from twenty farms vaccinated in 2016 and re-vaccinated in 2017 were included in the study. Two farms from the same area with good calving management were selected for investigation of passive LSDV antibody transfer from vaccinated mothers to new-borne calves.ResultsAll investigated cattle were healthy on the day of vaccination and during the whole study. Swelling at the injection site or other side effects of vaccination did not occur after re-vaccination in the study.Detection of LSD-specific antibodies was performed with the standard serological methods VNT and IFAT as well as a commercially available Capripox double antigen multi-species-ELISA. Capripoxvirus-specific antibodies were detected 46 to 47weeks after vaccination in 2016, with VNT in 35.06% and with IFAT and ELISA in 33.77%. A secondary response was observed in all three tests 1 month after re-vaccination with a significant increase in seropositive animals compared to the results before re-vaccination. With all applied serological methods, the number of animals testing positive was significantly higher at 1 and 5 months post re-vaccination than before re-vaccination. No significant statistical difference (p>0.05) was observed between the results of all three tests used. The sensitivity and specificity of ELISA was estimated to be Se-ELISA 91% and Sp(ELISA) 87% calculated by the results of VNT and Se-ELISA 88% and Sp(ELISA) 76% calculated by the results of IFAT. Passive antibody transfer from vaccinated mothers to new-born calves was investigated at 14days after birth. Discrepancies for the detection of LSDV specific antibodies between cows and newborn calves at the age of 14days were observed in VNT and IFAT, but not in ELISA.ConclusionOf all tests used the commercially available ELISA shows to be the most useful for high throughput analysis compared to VNT or IFAT.
PB  - BMC, London
T2  - BMC Veterinary Research
T1  - Humoral immune response to repeated lumpy skin disease virus vaccination and performance of serological tests
VL  - 15
SP  - 80
DO  - 10.1186/s12917-019-1831-y
ER  - 

@article{
author = "Milovanović, Milovan and Dietze, Klaas and Milićević, Vesna and Radojičić, Sonja and Valčić, Miroslav and Moritz, Tom and Hoffmann, Bernd",
year = "2019",
abstract = "BackgroundIn the presented study we investigated the development of the humoral immune response against LSDV during the process of re-vaccination of cattle over a time span of 5 months. In addition, the performance of different serological techniques for antibody detection against LSDV was compared. For sample collection, an area without previous LSD outbreak reports in Serbia was selected. Seventy-nine cattle from twenty farms vaccinated in 2016 and re-vaccinated in 2017 were included in the study. Two farms from the same area with good calving management were selected for investigation of passive LSDV antibody transfer from vaccinated mothers to new-borne calves.ResultsAll investigated cattle were healthy on the day of vaccination and during the whole study. Swelling at the injection site or other side effects of vaccination did not occur after re-vaccination in the study.Detection of LSD-specific antibodies was performed with the standard serological methods VNT and IFAT as well as a commercially available Capripox double antigen multi-species-ELISA. Capripoxvirus-specific antibodies were detected 46 to 47weeks after vaccination in 2016, with VNT in 35.06% and with IFAT and ELISA in 33.77%. A secondary response was observed in all three tests 1 month after re-vaccination with a significant increase in seropositive animals compared to the results before re-vaccination. With all applied serological methods, the number of animals testing positive was significantly higher at 1 and 5 months post re-vaccination than before re-vaccination. No significant statistical difference (p>0.05) was observed between the results of all three tests used. The sensitivity and specificity of ELISA was estimated to be Se-ELISA 91% and Sp(ELISA) 87% calculated by the results of VNT and Se-ELISA 88% and Sp(ELISA) 76% calculated by the results of IFAT. Passive antibody transfer from vaccinated mothers to new-born calves was investigated at 14days after birth. Discrepancies for the detection of LSDV specific antibodies between cows and newborn calves at the age of 14days were observed in VNT and IFAT, but not in ELISA.ConclusionOf all tests used the commercially available ELISA shows to be the most useful for high throughput analysis compared to VNT or IFAT.",
publisher = "BMC, London",
journal = "BMC Veterinary Research",
title = "Humoral immune response to repeated lumpy skin disease virus vaccination and performance of serological tests",
volume = "15",
pages = "80",
doi = "10.1186/s12917-019-1831-y"
}

Milovanović, M., Dietze, K., Milićević, V., Radojičić, S., Valčić, M., Moritz, T.,& Hoffmann, B.. (2019). Humoral immune response to repeated lumpy skin disease virus vaccination and performance of serological tests. in BMC Veterinary Research
BMC, London., 15, 80.
https://doi.org/10.1186/s12917-019-1831-y

Milovanović M, Dietze K, Milićević V, Radojičić S, Valčić M, Moritz T, Hoffmann B. Humoral immune response to repeated lumpy skin disease virus vaccination and performance of serological tests. in BMC Veterinary Research. 2019;15:80.
doi:10.1186/s12917-019-1831-y .

Milovanović, Milovan, Dietze, Klaas, Milićević, Vesna, Radojičić, Sonja, Valčić, Miroslav, Moritz, Tom, Hoffmann, Bernd, "Humoral immune response to repeated lumpy skin disease virus vaccination and performance of serological tests" in BMC Veterinary Research, 15 (2019):80,
https://doi.org/10.1186/s12917-019-1831-y . .