Molecular Characterisation and Antibody Response to Bovine Respiratory Syncytial Virus in Vaccinated and Infected Cattle in Turkey
Abstract
:1. Introduction
2. Materials and Methods
2.1. Study Design
2.2. Sampling and Study Population
2.3. Extraction of RNA and Reverse Transcription
2.4. Determination of RNA Concentration and Purity
2.5. PCR Inhibitors of RNA Extracts
2.6. Detection of BRSV by Real Time RT-PCR
2.7. Sequencing and Phylogenetic Analysis
2.8. ELISA
2.9. Virus Isolation
2.10. Statistical Analyses
3. Results
3.1. Clinical and Necropsy Findings
3.2. Seropositivity of BRSV Detected by ELISA
3.3. Efficiency of Real Time RT-PCR and Detection of BRSV-RNA in Clinical Samples
3.4. Sequencing and Phylogenetic Analysis
3.5. Virus Isolation
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Sample Collection Date | Sample Region | Age (Months) | Number of Samples | Clinical Signs/Pathology | ||
---|---|---|---|---|---|---|
Blood | Swabs | Lungs | ||||
21 January 2019 | Luleburgaz | 0–6 | 10 | 10 | Dyspnea, cough, serous nasal secretions, fatigue | |
20 February 2019 | Luleburgaz | 0–6 | 2 | 2 | Dyspnea, cough, serous nasal secretions, fatigue | |
20 February 2019 | Catalca-Istanbul | 0–6 | 18 | Dyspnea, cough, serous nasal secretions, fatigue | ||
21 February 2019 | Luleburgaz | 0–6 | 11 | 11 | 2 | Dead calf |
26 February 2019 | Usak | 0–6 | 20 | Dyspnea, cough, serous nasal secretions, fatigue | ||
13 January 2019 | Malkara | 0–6 | 18 | 18 | Dyspnea, cough, serous nasal secretions, fatigue | |
14 January 2019 | Kirklareli | 0–3 | 9 | 9 | Cough, serous nasal secretions, fatigue | |
26 January 2019 | Kirklareli | 0–6 | 25 | Dyspnea, cough, serous nasal secretions, fatigue | ||
10 February 2019 | Edirne | 0–3 | 15 | 15 | Cough, serous nasal secretions, fatigue | |
13 February 2019 | Cilingirkoy-Istanbul | 0–12 | 11 | 10 | Fever and cough | |
22 February 2019 | Luleburgaz | 0–6 | 19 | Dyspnea, cough, serous nasal secretions, fatigue | ||
23 February 2019 | Kirklareli | 0–12 | 3 | 1 | Dyspnea, cough, serous nasal secretions, fatigue | |
24 February 2019 | Kirklareli | 0–6 | 5 | Dyspnea, cough, serous nasal secretions, fatigue | ||
4 March 2021 | Luleburgaz | 0–6 | 25 | 25 | 1 | Dead calf |
9 March 2021 | Kirklareli | 0–6 | 4 | Dyspnea, cough, serous nasal secretions, fatigue | ||
18 March 2021 | Kirklareli | 0–6 | 9 | 9 | Dyspnea, cough, serous nasal secretions, fatigue | |
2 April 2021 | Kirklareli | 0–6 | 5 | Dyspnea, cough, serous nasal secretions, fatigue | ||
2 April 2021 | Buyukcekmece-Istanbul | 6–18 | 5 | Abattoir sample: Consolidation, hyperemia | ||
7 April 2021 | Cilingirkoy-Istanbul | 0–12 | 7 | 7 | Fever and cough | |
19 April 2021 | Edirne | 0–12 | 12 | 12 | Cough, serous nasal secretions, fatigue | |
7 April 2021 | Balikesir | 12–24 | 19 | Abattoir sample: Consolidation, hyperemia | ||
4 November 2021 | Edirne | 0–6 | 6 | 6 | Cough, serous nasal secretions, fatigue | |
17 December 2021 | Edirne | 0–6 | 11 | 11 | Cough, serous nasal secretions, fatigue | |
5 January 2022 | Esenyurt-Istanbul | 12–24 | 9 | Abattoir sample: Consolidation, hyperemia | ||
20 January 2022 | Usak | 0–6 | 3 | 3 | 1 | Dead calf Cough, serous nasal secretions, fatigue |
1 March 2022 | Edirne | 0–3 | 2 | Cough, serous nasal secretions, fatigue | ||
4 March 2022 | Usak | 0–6 | 20 | Cough, serous nasal secretions, fatigue | ||
7 March 2022 | Kirklareli | 0–3 | 10 | 10 | Fever, serous nasal secretions, fatigue | |
Total | 162 | 277 | 37 |
Primers and Probe | Target Genes | Primers and Probe Sequences | Size (bp) | Positions | References |
---|---|---|---|---|---|
B5A-B6A RT-PCR | G gene | F: 5′-CCACCCTAGCAATGATAACCTTGAC-3′ R: 5′-AAGAGAGGATGC(T/C)TTGCTGTGG-3′ | 603 | 110–134 691–712 | [29] |
BRSV Real time RT-PCR | N gene | F: 5′-GCAATGCTGCAGGACTAGGTATAAT-3′ R: 5′-ACACTGTAATTGATGACCCCATTCT-3′ | 123 | 977–1001 1076–1100 | [39] |
Probe | FAM-5-ACCAAGACTTGTATGATGCTGCCAAAGCA-3-TAMRA | 1028–1056 | [39] |
Distance/ Distance | OQ713830_9716 2019_TÜRKİYE | OQ689745_7578 2019_TÜRKİYE | OQ736254_USOS1 2022_TÜRKİYE | OQ743526_USOS3 2022_TÜRKİYE | OQ789520_USAKC 2022_TÜRKİYE | Distance/ Distance | OQ713830_9716 2019_TÜRKİYE | OQ689745_7578 2019_TÜRKİYE | OQ736254_USOS1 2022_TÜRKİYE | OQ743526_USOS3 2022_TÜRKİYE | OQ789520_USAKC 2022_TÜRKİYE |
---|---|---|---|---|---|---|---|---|---|---|---|
OQ713830_9716_2019_TÜRKİYE | 0.00 | 96.36 | 89.88 | 89.88 | 88.26 | KY753468.1_2017_ITALY | 87.85 | 87.04 | 87.04 | 87.04 | 85.43 |
OQ689745_7578_2019_TÜRKİYE | 96.36 | 0.00 | 89.07 | 89.07 | 87.45 | M58307.1_1990_USA | 87.45 | 87.45 | 86.64 | 86.64 | 85.02 |
AY910755.1_2005_CZECHIA | 92.71 | 93.12 | 91.50 | 91.50 | 89.88 | Y08719.1_1997_ENGLAND | 87.45 | 88.26 | 87.45 | 87.45 | 85.83 |
L08414.1_1997_USA | 92.71 | 93.12 | 90.69 | 90.69 | 89.47 | AY910765.1_2005_CZECHIA | 87.04 | 85.83 | 87.04 | 87.04 | 85.43 |
AY910756.1_2005_CZECHIA | 92.31 | 92.71 | 91.09 | 91.09 | 89.47 | KF501149.1_2014_NORWAY | 87.04 | 87.85 | 86.23 | 86.23 | 84.62 |
L08416.1_1997_USA | 91.90 | 92.31 | 90.69 | 90.69 | 89.07 | FJ555202.1_2008_BRASIL | 87.04 | 86.23 | 86.23 | 86.23 | 84.62 |
L08415.1_1997_USA | 91.90 | 92.31 | 90.69 | 90.69 | 89.07 | AF188583.1_2000_FRANCE | 87.04 | 87.04 | 87.04 | 87.04 | 85.43 |
MW881233.1_2021_TÜRKİYE | 91.09 | 91.90 | 93.93 | 93.93 | 92.31 | Y08717.1_1997_ENGLAND | 86.64 | 86.23 | 86.64 | 86.64 | 85.02 |
U24716.1_1997_BELGIUM | 91.09 | 91.50 | 90.69 | 90.69 | 89.07 | U92100.1_1997_DENMARK | 86.23 | 86.23 | 86.23 | 86.23 | 84.62 |
LC499991.1_2019_JAPAN | 91.09 | 92.31 | 89.88 | 89.88 | 88.26 | AF188588.1_2000_FRANCE | 85.83 | 85.43 | 85.83 | 85.83 | 84.21 |
L08411.1_1997_USA | 90.69 | 91.09 | 89.47 | 89.47 | 87.85 | KY660261.1_2017_CROATIA | 85.43 | 85.43 | 83.81 | 83.81 | 82.19 |
MH133326.1_2018_TÜRKIYE | 90.28 | 89.47 | 98.79 | 98.79 | 97.17 | KY753469.1_2017_ITALY | 85.43 | 83.81 | 84.62 | 84.62 | 83.00 |
MH133327.1_2018_TÜRKİYE | 90.28 | 89.47 | 98.79 | 98.79 | 97.17 | AF188604.1_2000_FRANCE | 85.43 | 85.02 | 86.23 | 86.23 | 84.62 |
OQ736254_USOS1_2022_TÜRKİYE | 89.88 | 89.07 | 0.00 | 100.00 | 98.38 | AF188603.1_2000_FRANCE | 84.62 | 84.21 | 85.43 | 85.43 | 83.81 |
OQ743526_USOS3_2022_TÜRKİYE | 89.88 | 89.07 | 100.00 | 0.00 | 98.38 | KY753463.1_2017_ITALY | 84.62 | 85.02 | 83.81 | 83.81 | 82.19 |
MW881234.1_2021_TÜRKiYE | 89.88 | 90.69 | 95.14 | 95.14 | 93.52 | KF501172.1_2014_NORWAY | 84.21 | 85.02 | 82.59 | 82.59 | 80.97 |
KY680331.1_2018_CROATIA | 89.88 | 88.66 | 88.66 | 88.66 | 87.04 | KY753456.1_2017_ITALY | 84.21 | 83.81 | 85.02 | 85.02 | 83.40 |
LC499984.1_2019_JAPAN | 89.88 | 90.28 | 89.47 | 89.47 | 87.85 | KY753464.1_2017_ITALY | 84.21 | 83.81 | 83.00 | 83.00 | 81.38 |
KY680337.1_2018_CROATIA | 89.47 | 88.26 | 88.26 | 88.26 | 86.64 | KY680318.1_2018_CROATIA | 84.21 | 84.62 | 84.21 | 84.21 | 82.59 |
KY680336.1_2018_CROATIA | 89.47 | 88.26 | 88.26 | 88.26 | 86.64 | KF501153.1_2014_NORWAY | 83.81 | 85.43 | 82.19 | 82.19 | 80.57 |
KY680333.1_2018_CROATIA | 89.47 | 88.26 | 88.26 | 88.26 | 86.64 | AF188597.1_2000_FRANCE | 83.81 | 83.40 | 83.81 | 83.81 | 82.19 |
KY753451.1_2017_ITALY | 89.47 | 88.26 | 88.26 | 88.26 | 86.64 | MK599397.1_2019_BRASIL | 83.81 | 84.21 | 82.59 | 82.59 | 80.97 |
MN129181.1_2019_IRAQ | 89.07 | 87.85 | 87.85 | 87.85 | 86.23 | KY680327.1_2018_CROATIA | 83.40 | 83.81 | 84.21 | 84.21 | 82.59 |
AF092942.1_1998_GERMANY | 89.07 | 89.07 | 87.45 | 87.45 | 85.83 | KY680328.1_2018_CROATIA | 83.00 | 83.40 | 83.81 | 83.81 | 82.19 |
KY753452.1_2017_ITALY | 89.07 | 87.85 | 88.66 | 88.66 | 87.04 | Y11205.1_1997_ENGLAND | 82.59 | 81.38 | 81.38 | 81.38 | 79.76 |
LC499985.1_2019_JAPAN | 89.07 | 88.66 | 87.85 | 87.85 | 86.23 | U57823.1_1997_BELGIUM | 82.59 | 81.38 | 81.38 | 81.38 | 79.76 |
OQ789520_USAKC_2022_TÜRKİYE | 88.26 | 87.45 | 98.38 | 98.38 | 0.00 | AF295544.1_2001_USA | 82.59 | 81.38 | 81.38 | 81.38 | 79.76 |
AF248593.1_2000_DENMARK | 88.26 | 88.26 | 87.45 | 87.45 | 85.83 | MK599389.1_2019_BRASIL | 82.19 | 83.40 | 80.57 | 80.57 | 78.95 |
AF188584.1_2000_FRANCE | 88.26 | 88.26 | 87.45 | 87.45 | 85.83 | AF188585.1_2000_FRANCE | 81.38 | 81.78 | 82.19 | 82.19 | 80.57 |
AF188582.1_2000_FRANCE | 88.26 | 88.26 | 86.64 | 86.64 | 85.02 | AF188586.1_2000_FRANCE | 81.38 | 81.78 | 82.19 | 82.19 | 80.57 |
LC499994.1_2019_JAPAN | 88.26 | 88.66 | 88.66 | 88.66 | 87.04 | AF188587.1_2000_FRANCE | 81.38 | 81.78 | 82.19 | 82.19 | 80.57 |
U24714.1_1997_BELGIUM | 87.85 | 87.85 | 87.04 | 87.04 | 85.43 |
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Aydin, O.; Yilmaz, A.; Turan, N.; Richt, J.A.; Yilmaz, H. Molecular Characterisation and Antibody Response to Bovine Respiratory Syncytial Virus in Vaccinated and Infected Cattle in Turkey. Pathogens 2024, 13, 304. https://doi.org/10.3390/pathogens13040304
Aydin O, Yilmaz A, Turan N, Richt JA, Yilmaz H. Molecular Characterisation and Antibody Response to Bovine Respiratory Syncytial Virus in Vaccinated and Infected Cattle in Turkey. Pathogens. 2024; 13(4):304. https://doi.org/10.3390/pathogens13040304
Chicago/Turabian StyleAydin, Ozge, Aysun Yilmaz, Nuri Turan, Juergen A. Richt, and Huseyin Yilmaz. 2024. "Molecular Characterisation and Antibody Response to Bovine Respiratory Syncytial Virus in Vaccinated and Infected Cattle in Turkey" Pathogens 13, no. 4: 304. https://doi.org/10.3390/pathogens13040304
APA StyleAydin, O., Yilmaz, A., Turan, N., Richt, J. A., & Yilmaz, H. (2024). Molecular Characterisation and Antibody Response to Bovine Respiratory Syncytial Virus in Vaccinated and Infected Cattle in Turkey. Pathogens, 13(4), 304. https://doi.org/10.3390/pathogens13040304