Spleen Swabs for Sensitive and High-Throughput Detection of African Swine Fever Virus by Real-Time PCR
Abstract
:1. Introduction
2. Materials and Methods
2.1. Sample Origins
2.2. Preparation of Homogenates and Swabs from Spleen Tissue Samples
2.3. Real-Time PCR Detection of ASFV Genomic DNA, β-Actin and Armored Enterovirus RNA
2.4. Virus Isolation
2.5. Virus Titration
2.6. Statistical Analysis
3. Results and Discussion
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Strain | Sample | dpi | Tignon-ASFV | Zsak-ASFV | Moniwa-B-Actin | |||
---|---|---|---|---|---|---|---|---|
Homogenate | Swab | Homogenate | Swab | Homogenate | Swab | |||
ASFV Malta’78 | 1 | 3 | 23.15 | 23.47 | 20.78 | 21.13 | 21.08 | 18.32 |
2 | 4 | 19.53 | 18.21 | 17.26 | 17.08 | 21.66 | 20.64 | |
3 | 5 | 18.35 | 17.80 | 16.60 | 15.98 | 22.24 | 20.86 | |
4 | 3 | 28.28 | 27.74 | 25.74 | 25.43 | 21.40 | 20.44 | |
5 | 5 | 17.55 | 16.68 | 15.76 | 15.17 | 22.72 | 21.62 | |
6 | 2 | ND | 37.21 | 37.19 | 35.08 | 22.23 | 19.18 | |
7 | 4 | 20.05 | 19.39 | 18.25 | 17.42 | 22.13 | 21.47 | |
8 | 1 | ND | ND | ND | ND | 22.09 | 19.30 | |
9 | 4 | 18.83 | 18.36 | 16.39 | 16.29 | 22.10 | 20.72 | |
10 | 1 | ND | 39.88 | ND | 39.28 | 21.66 | 20.00 | |
11 | 3 | 27.92 | 26.40 | 25.52 | 24.32 | 21.11 | 20.04 | |
12 | 2 | ND | ND | ND | ND | 22.04 | 19.79 | |
13 | 1 | ND | ND | ND | ND | 21.07 | 20.25 | |
14 | 4 | 20.16 | 19.24 | 18.17 | 17.25 | 22.34 | 21.53 | |
15 | 3 | 27.43 | 27.50 | 25.04 | 24.88 | 22.97 | 19.95 | |
ASFV Georgia 2007/1 | 16 | 18 | 36.46 | 32.95 | 35.36 | 31.24 | 19.70 | 17.57 |
17 | 18 | 22.97 | 22.57 | 21.53 | 21.00 | 20.28 | 17.33 | |
18 | 17 | 16.66 | 16.58 | 15.85 | 16.01 | 22.05 | 21.67 | |
19 | 16 | 18.33 | 17.73 | 17.12 | 17.16 | 22.71 | 21.17 | |
20 | 6 | 17.64 | 20.48 | 17.06 | 19.20 | 23.19 | 22.85 | |
21 | 11 | 18.76 | 20.92 | 17.90 | 19.32 | 23.16 | 22.79 | |
22 | 18 | 19.32 | 20.36 | 18.33 | 19.08 | 23.32 | 22.53 | |
23 | 10 | 19.07 | 20.01 | 17.88 | 18.69 | 24.06 | 23.28 | |
24 | 18 | 17.83 | 18.89 | 16.76 | 17.45 | 23.26 | 21.69 | |
25 | 18 | 25.09 | 24.96 | 24.46 | 24.85 | 22.26 | 19.20 | |
26 | 22 | 17.37 | 19.14 | 16.63 | 17.73 | 23.87 | 23.34 | |
27 | 25 | ND | 35.21 | ND | 34.76 | 22.35 | 21.29 | |
28 | 25 | ND | 35.64 | ND | 33.74 | 21.68 | 21.12 | |
29 | 22 | 17.81 | 18.87 | 17.30 | 18.05 | 25.13 | 24.77 | |
30 | 25 | ND | 37.13 | ND | 36.25 | 21.22 | 20.53 | |
31 | 25 | ND | 37.54 | ND | 36.38 | 22.01 | 20.15 | |
32 | 25 | 18.44 | 19.81 | 17.27 | 18.66 | 25.57 | 22.72 | |
ASFVE stonia/2014 | 33 | 5 | ND | ND | ND | 38.13 | 22.62 | 19.01 |
34 | 11 | ND | ND | ND | ND | 22.55 | 20.00 | |
35 | 22 | ND | ND | 39.47 | ND | 22.06 | 20.93 | |
36 | 37 | ND | ND | ND | ND | 22.18 | 21.48 | |
37 | 6 | 34.18 | 34.13 | 32.05 | 31.71 | 22.17 | 20.16 | |
38 | 20 | 31.89 | 30.40 | 29.99 | 27.76 | 22.23 | 19.94 | |
39 | 26 | 37.80 | 36.66 | 35.39 | 33.22 | 22.42 | 20.40 | |
40 | 33 | 32.96 | 33.08 | 30.66 | 30.47 | 21.82 | 19.69 | |
41 | 1 | 19.27 | 20.48 | 17.25 | 17.63 | 21.79 | 20.25 | |
42 | 14 | 19.69 | 19.04 | 17.30 | 16.95 | 22.59 | 21.69 | |
43 | 30 | 18.68 | 17.14 | 16.65 | 15.23 | 23.90 | 22.93 | |
44 | 39 | 18.99 | 19.25 | 16.91 | 16.86 | 23.18 | 21.47 | |
45 | 2 | 19.82 | 18.85 | 17.63 | 16.62 | 22.41 | 21.03 | |
46 | 15 | 19.73 | 18.55 | 17.69 | 16.25 | 23.10 | 21.46 | |
47 | 27 | 19.39 | 18.61 | 17.50 | 16.95 | 22.87 | 21.03 | |
48 | 34 | 19.28 | 18.42 | 17.73 | 16.95 | 24.22 | 23.38 | |
49 | 3 | 17.81 | 17.57 | 16.19 | 15.84 | 23.48 | 22.94 | |
50 | 13 | 18.96 | 18.02 | 17.13 | 16.31 | 23.52 | 23.23 | |
51 | 29 | 17.29 | 17.85 | 15.70 | 15.66 | 24.07 | 22.98 | |
52 | 36 | 17.31 | 17.26 | 15.44 | 15.34 | 24.22 | 23.56 | |
Percent (%) Detection | 75.0 | 86.5 | 76.9 | 88.5 | 100.0 | 100.0 |
Strain | Sample | Tignon-ASFV | Zsak-ASFV | Moniwa-B-Actin | |||
---|---|---|---|---|---|---|---|
Homogenate | Swab | Homogenate | Swab | Homogenate | Swab | ||
ASFV Georgia 2007/1 | 16 | 38.27 | 33.63 | 35.06 | 30.88 | 19.36 | 18.46 |
17 | 22.41 | 22.62 | 21.34 | 20.66 | 19.91 | 18.22 | |
18 | 16.38 | 16.55 | 16.03 | 15.84 | 21.36 | 21.44 | |
26 | 17.97 | 18.88 | 17.02 | 17.73 | 23.07 | 22.83 | |
27 | ND | 34.07 | ND | 34.04 | 21.13 | 21.96 | |
28 | ND | 36.51 | 38.39 | 32.77 | 20.45 | 20.46 | |
29 | 19.02 | 18.88 | 18.00 | 17.92 | 24.24 | 24.07 | |
30 | 39.59 | 38.03 | ND | 34.56 | 19.98 | 20.46 | |
31 | 39.53 | 36.42 | ND | 37.16 | 20.52 | 21.04 | |
32 | 16.02 | 20.26 | 15.635 | 19.04 | 24.75 | 23.22 | |
Percent (%) Detection | 80.0 | 100.0 | 70.0 | 100.0 | 100.0 | 100.0 |
Sample | Tignon-ASF | Titer-Log (TCID50/mL) | ||
---|---|---|---|---|
Homogenate | Swab | Homogenate | Swab | |
11 | 27.92 | 26.40 | 4.10 | 4.87 |
18 | 16.66 | 16.58 | 7.53 | 8.65 |
38 | 31.89 | 30.40 | 3.97 | 4.80 |
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Cafariello, C.; Goonewardene, K.; Chung, C.J.; Ambagala, A. Spleen Swabs for Sensitive and High-Throughput Detection of African Swine Fever Virus by Real-Time PCR. Viruses 2024, 16, 1316. https://doi.org/10.3390/v16081316
Cafariello C, Goonewardene K, Chung CJ, Ambagala A. Spleen Swabs for Sensitive and High-Throughput Detection of African Swine Fever Virus by Real-Time PCR. Viruses. 2024; 16(8):1316. https://doi.org/10.3390/v16081316
Chicago/Turabian StyleCafariello, Christopher, Kalhari Goonewardene, Chungwon J. Chung, and Aruna Ambagala. 2024. "Spleen Swabs for Sensitive and High-Throughput Detection of African Swine Fever Virus by Real-Time PCR" Viruses 16, no. 8: 1316. https://doi.org/10.3390/v16081316