Molecular Characterization and Developing a Point-of-Need Molecular Test for Diagnosis of Bovine Papillomavirus (BPV) Type 1 in Cattle from Egypt
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Ethical Statement
2.2. Animals
2.3. Surgical Treatment of the Cutaneous Warts and Sampling
2.4. Skin Wart Sample Preparations
2.5. Polymerase Chain Reaction (PCR)
2.6. PCR Product Sequencing and Analysis
2.7. Developing Recombinase Polymerase Amplification-Nucleic Acid Lateral Flow Immunoassays (RPA-NALF) for Point-of-Need Molecular Identification of BPV
2.7.1. Oligonucleotide Primers for RPA
2.7.2. BPV-1 RPA Amplification
2.7.3. Visualization with Nucleic Acid Lateral Flow (NALF) Strip
2.7.4. The Limit of Detection (LOD)
2.7.5. BPV RPA-NALF Immunoassay Detection Performance
3. Results
3.1. PCR Detection and Sequence Analysis
3.2. RPA-NALF Assay Results
4. Discussion
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Farm Number | Age Range (Year) | Total Number of Animals/Farm | Sex of Examined Animals | No, Sex, and Age of Animals Showed Clinical Signs and Positive Result by BPV-1-PCR | ||
---|---|---|---|---|---|---|
Female | Male | |||||
1 | 1–4 | 15 | 10 | 5 | 0 | |
2 | 1–5 | 19 | 11 | 8 | 1 male (13 month) | |
3 | 1–3 | 13 | 9 | 4 | 0 | |
4 | 1–5 | 21 | 13 | 8 | 1 Female (18 months) 1 male (24 month) | |
5 | 1–5 | 16 | 10 | 6 | 0 | |
6 | 1–3 | 10 | 8 | 2 | 1 male (23 month) | |
7 | 1–4 | 15 | 11 | 4 | 0 | |
8 | 1–3 | 18 | 12 | 6 | 0 | |
9 | 1–3 | 12 | 8 | 4 | 0 | |
10 | 1–5 | 14 | 10 | 4 | 1 Female (14 month) | |
11 | 1–4 | 16 | 11 | 5 | 1 Female (13 month) | |
12 | 1–5 | 15 | 10 | 5 | 0 | |
13 | 1–3 | 20 | 12 | 8 | 0 | |
14 | 1–4 | 14 | 10 | 4 | 1 Female (20 month) | |
15 | 1–4 | 17 | 11 | 6 | 1 Female (15 month) | |
16 | 1–5 | 10 | 8 | 2 | 0 | |
17 | 1–4 | 19 | 12 | 7 | 1 Female (14 month) | |
18 | 1–5 | 19 | 13 | 6 | 0 | |
19 | 1–3 | 12 | 9 | 3 | 1 Female (17 month) | |
20 | 1–5 | 13 | 9 | 4 | 1 Female (22 month) | |
Total (%) | 308 (100%) | 207 (67.2%) | 101 (32.8%) | 11 (3.6%) | ||
8 females (2.6%) | 3 males (1.0%) |
Isolate Number | Country of Isolation | Year of Identification | Host Species | Accession Number |
---|---|---|---|---|
BPV 1 | Egypt\New Valley Governorate (This study) | 2016 | Cattle | MH543316 |
BPV 2 | India | 2014 | Cattle | HG918265 |
BPV 3 | Sweden | 1983 | Cattle | J02045 |
BPV 4 | China | 2017 | Cattle | MF045489 |
BPV 5 | Morocco | 2017 | Cattle | KY746722 |
BPV 6 | India | 2012 | Buffalo | KF148690 |
BPV 7 | India | 2012 | Equine | KF114855 |
BPV 8 | Japan | 2011 | Cattle | AB626705 |
BPV 9 | Switzerland | 2017 | Cattle | MF384294 |
BPV 10 | Switzerland | 2017 | Cattle | MF384293 |
BPV 11 | Switzerland | 2017 | Cattle | MF384292 |
BPV 12 | Switzerland | 2017 | Cattle | MF384291 |
BPV 13 | Switzerland | 2017 | Cattle | MF384290 |
BPV 14 | India | 2018 | Cattle | MK396096 |
BPV 15 | Japan | 2016 | Cattle | LC426023 |
BPV 16 | India | 2018 | Cattle | MK173052 |
BPV 17 | Turkey | 2018 | Cattle | MH197482 |
BPV 18 | Japan | 2014 | Cattle | LC333380 |
BPV 19 | USA | 2012 | Equine | KY886226 |
BPV 20 | China | 2018 | Cattle | MK347523 |
BPV 21 | China | 2017 | Cattle | MG263871 |
BPV 22 | China | 2016 | Cattle | MF435917 |
BPV 23 | China | 2016 | Cattle | MF435916 |
BPV 24 | China | 2016 | Cattle | KX907623 |
BPV 25 | Switzerland | 2017 | Equine | MF384284 |
BPV 26 | Switzerland | 2017 | Equine | MF384283 |
BPV 27 | Switzerland | 2017 | Equine | MF384282 |
BPV 28 | Switzerland | 2017 | Equine | MF384286 |
BPV 29 | Croatia | 2010 | Cattle | JX046521 |
BPV 30 | India | 2017 | Cattle | LT837966 |
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El-Tholoth, M.; Mauk, M.G.; Elnaker, Y.F.; Mosad, S.M.; Tahoun, A.; El-Sherif, M.W.; Lokman, M.S.; Kassab, R.B.; Abdelsadik, A.; Saleh, A.A.; et al. Molecular Characterization and Developing a Point-of-Need Molecular Test for Diagnosis of Bovine Papillomavirus (BPV) Type 1 in Cattle from Egypt. Animals 2020, 10, 1929. https://doi.org/10.3390/ani10101929
El-Tholoth M, Mauk MG, Elnaker YF, Mosad SM, Tahoun A, El-Sherif MW, Lokman MS, Kassab RB, Abdelsadik A, Saleh AA, et al. Molecular Characterization and Developing a Point-of-Need Molecular Test for Diagnosis of Bovine Papillomavirus (BPV) Type 1 in Cattle from Egypt. Animals. 2020; 10(10):1929. https://doi.org/10.3390/ani10101929
Chicago/Turabian StyleEl-Tholoth, Mohamed, Michael G. Mauk, Yasser F. Elnaker, Samah M. Mosad, Amin Tahoun, Mohamed W. El-Sherif, Maha S. Lokman, Rami B. Kassab, Ahmed Abdelsadik, Ayman A. Saleh, and et al. 2020. "Molecular Characterization and Developing a Point-of-Need Molecular Test for Diagnosis of Bovine Papillomavirus (BPV) Type 1 in Cattle from Egypt" Animals 10, no. 10: 1929. https://doi.org/10.3390/ani10101929