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Evaluation of various filter paper and reagent systems for the preservation of Newcastle disease virus RNA samples

Bajes Amjed Al Qaisieh

Bajes Amjed Al Qaisieh

Department of Basic Medical Veterinary Sciences, Faculty of Veterinary Medicine, Jordan University of Science and TechnologyIrbid, Jordan
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Qaisieh, Bajes Amjed Al
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Mustafa Mohammed-Khair Ababneh

Mustafa Mohammed-Khair Ababneh

Department of Basic Medical Veterinary Sciences, Faculty of Veterinary Medicine, Jordan University of Science and TechnologyIrbid, Jordan
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Ababneh, Mustafa Mohammed-Khair
, 
Mohammad Borhan F. Al-Zghoul

Mohammad Borhan F. Al-Zghoul

Department of Basic Medical Veterinary Sciences, Faculty of Veterinary Medicine, Jordan University of Science and TechnologyIrbid, Jordan
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Al-Zghoul, Mohammad Borhan F.
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Daoud Abed Alnaser Alghizzawi

Daoud Abed Alnaser Alghizzawi

Department of Basic Medical Veterinary Sciences, Faculty of Veterinary Medicine, Jordan University of Science and TechnologyIrbid, Jordan
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Alghizzawi, Daoud Abed Alnaser
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Hebah Alaeddin Aboomer

Hebah Alaeddin Aboomer

Department of Clinical Medical Veterinary Sciences, Faculty of Veterinary Medicine, Jordan University of Science and TechnologyIrbid, Jordan
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Aboomer, Hebah Alaeddin
  
09 jun 2025
Evaluation of various filter paper and reagent systems for the preservation of Newcastle disease virus RNA samples's Cover Image
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Publicado en línea: 09 jun 2025
Páginas: 183 - 190
Recibido: 21 dic 2024
Aceptado: 20 may 2025
DOI: https://doi.org/10.2478/jvetres-2025-0030
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© 2025 Bajes Amjed Al Qaisieh et al., published by Sciendo
This work is licensed under the Creative Commons Attribution 4.0 International License.

Fig. 1.

Study methodology. Newcastle disease virus (NDV) RNA preservation using various methods (Flinders Technology Associates (FTA) cards, RNASound cards and RNAstable tubes) across temperatures (–80°C, –20°C, 4°C, 25°C and 56°C) and incubation periods (1, 7, 14, 28 and 35 d). The quality of RNA was assessed using real-time reverse-transcription (RT)-PCR, ratiometric fluorometry and Sanger sequencing
Study methodology. Newcastle disease virus (NDV) RNA preservation using various methods (Flinders Technology Associates (FTA) cards, RNASound cards and RNAstable tubes) across temperatures (–80°C, –20°C, 4°C, 25°C and 56°C) and incubation periods (1, 7, 14, 28 and 35 d). The quality of RNA was assessed using real-time reverse-transcription (RT)-PCR, ratiometric fluorometry and Sanger sequencing

Fig. 2.

Values of Δ threshold cycle (ΔCt) for Newcastle virus RNA samples preserved with Flinders Technology Associates (FTA) cards, RNASound cards and RNAstable tubes or not preserved as free virus and free RNA controls and incubated at –20°C, 4°C, 25°C and 56°C; ΔCts are compared to those of RNA samples stored at –80°C. A – day 1; B – day 7; C – day 14; D – day 28; E – day 35; * – excluded datum or undetectable amplification
Values of Δ threshold cycle (ΔCt) for Newcastle virus RNA samples preserved with Flinders Technology Associates (FTA) cards, RNASound cards and RNAstable tubes or not preserved as free virus and free RNA controls and incubated at –20°C, 4°C, 25°C and 56°C; ΔCts are compared to those of RNA samples stored at –80°C. A – day 1; B – day 7; C – day 14; D – day 28; E – day 35; * – excluded datum or undetectable amplification

Reference sequence to partial F gene sequence match results after Newcastle virus RNA preservation as free virus and free RNA and by Flinders Technology Associates (FTA) card, RNASound and RNAstable

Free virus FTA cards RNASound cards Free RNA RNAstable tubes
Sample number Treatment (°C)/days Sample number Treatment (°C)/days Sample number Treatment (°C)/days Sample number Treatment (°C)/days Sample number Treatment (°C)/days
1 –80/1 10 –80/1 19 –80/1 28 –80/1 37 –80/1
7 –80/35 11 25/1 20 25/1 29 25/1 38 25/1
8 25/35 12 56/1 21 56/1 30 56/1 39 56/1
- - 14 25/28 22 –80/28 31 –80/28 40 –80/28
- - 15 56/28 24 56/28 32 25/28 41 25/28
- - 16 –80/35 26 25/35 34 –80/35 42 56/28
- - 17 25/35 27 56/35 35 25/35 43 –80/35
- - - - - - - - 44 25/35
- - - - - - - - 45 56/35
Matching samples/total samples - 3/9 - 7/9 - 7/9 - 7/9 - 9/9

Newcastle virus RNA integrity data for various preservation methods and controls at three temperatures across three time points_ The integrity of RNA was scored in a range from 1 to 10, where 10 was the highest integrity

Temperature (°C) FTA cards RNASound cards RNAstable tubes Free virus Free RNA
–80 10 N.D. 9.1 9.4 8.7
Day 1 25 10 N.D. N.D. 9.0 9.5
56 10 N.D. N.D. 8.5 7.9
–80 10 N.D. 9.1 9.3 8.6
Day 14 25 10 N.D. N.D. 8.9 9.5
56 10 N.D. N.D. 8.3 7.4
–80 10 N.D. 9.2 8.6 8.6
Day 35 25 10 N.D. N.D. 8.9 9.4
56 10 N.D. N.D. 8.2 7.2
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Ciencias de la vida, Biología molecular, Microbiología y virología, Ciencias de la vida, otros, Medicina, Medicina veterinaria
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