Evidence for the type of nucleic acid in African swine fever virus

1966 ◽  
Vol 19 (3) ◽  
pp. 289-304 ◽  
Author(s):  
W. Plowright ◽  
F. Brown ◽  
J. Parker
2021 ◽  
Vol 22 (23) ◽  
pp. 12915
Author(s):  
Ahmed Elnagar ◽  
Timm C. Harder ◽  
Sandra Blome ◽  
Martin Beer ◽  
Bernd Hoffmann

FTA cards and related products simplify the collection, transport, and transient storage of biological sample fluids. Here, we have compared the yield and quality of DNA and RNA released from seven different FTA cards using seven releasing/extraction methods with eleven experimental eluates. For the validation, dilution series of African swine fever virus (ASFV) positive EDTA blood and Influenza A virus (IAV) positive allantoic fluid were used. Based on our data, we conclude that direct PCR amplification without the need for additional nucleic acid extraction and purification could be suitable and more convenient for ASFV DNA release from FTA cards. In contrast, IAV RNA loads can be amplified from FTA card punches if a standard extraction procedure including a lysis step is applied. These differences between the amplifiable viral DNA and RNA after releasing and extraction are not influenced by the type of commercial FTA card or the eleven different nucleic acid releasing procedures used for the comparative analyses. In general, different commercial FTA cards were successfully used for the storage and recovery of the ASFV and IAV genetic material suitable for PCR. Nevertheless, the usage of optimized nucleic acid releasing protocols could improve the recovery of the viral genome of both viruses. Here, the application of Chelex® Resin 100 buffer mixed with 1 × Tris EDTA buffer (TE, pH 8.0) or with TED 10 (TE buffer and Dimethylsulfoxid) delivered the best results and can be used as a universal method for releasing viral DNA and RNA from FTA cards.


2019 ◽  
Author(s):  
Qian He ◽  
Dongmei Yu ◽  
Mengdi Bao ◽  
Grant Korensky ◽  
Juhong Chen ◽  
...  

AbstractHere we report the development of a high throughput, all-solution phase, and isothermal detection system to detect African Swine Fever Virus (ASFV). CRISPR-Cas12a programmed with a CRISPR RNA (crRNA) is used to detect ASFV target DNA. Upon ASFV DNA binding, the Cas12a/crRNA/ASFV DNA complex becomes activated and degrades a fluorescent single stranded DNA (ssDNA) reporter present in the assay. We combine this powerful CRISPR-Cas assay with fluorescence-based point-of-care (POC) system we developed for rapid and accurate virus detection. Without nucleic acid amplification, a detection limit of 1 pM is achieved within 2 hrs. In addition, the ternary Cas12a/crRNA/ASFV DNA complex is highly stable at physiological temperature and continues to cleave the ssDNA reporter even after 24 hrs of incubation, resulting in an improvement of the detection limit to 100 fM. We show that this system is very specific and can differentiate nucleic acid targets with closely matched sequences. The high sensitivity and selectivity of our system enables the detection of ASFV in femtomolar range. Importantly, this system features a disposable cartridge and a sensitive custom designed fluorometer, enabling compact, multiplexing, and simple ASFV detection, intended for low resource settings.


2018 ◽  
Vol 261 ◽  
pp. 14-16 ◽  
Author(s):  
Ann Sofie Olesen ◽  
Louise Lohse ◽  
Marlene Danner Dalgaard ◽  
Grzegorz Woźniakowski ◽  
Graham J. Belsham ◽  
...  

2020 ◽  
Vol 23 (04) ◽  
pp. 21-26
Author(s):  
A.K. Sibgatullova ◽  
◽  
M.E. Vlasov ◽  
I.A. Titov ◽  
◽  
...  

2020 ◽  
Vol 23 (2) ◽  
pp. 14-19
Author(s):  
A.K. Sibgatullova ◽  
◽  
M.V. Nefedeva ◽  
D.A. Kudryashov ◽  
M.E. Vlasov ◽  
...  

1992 ◽  
Vol 66 (6) ◽  
pp. 3860-3868 ◽  
Author(s):  
A Alcamí ◽  
A Angulo ◽  
C López-Otín ◽  
M Muñoz ◽  
J M Freije ◽  
...  

1990 ◽  
Vol 64 (5) ◽  
pp. 2064-2072 ◽  
Author(s):  
J M Almendral ◽  
F Almazán ◽  
R Blasco ◽  
E Viñuela

2021 ◽  
pp. 105081
Author(s):  
Zhao Huang ◽  
Lang Gong ◽  
Zezhong Zheng ◽  
Qi Gao ◽  
Xiongnan Chen ◽  
...  

Author(s):  
Reet Herm ◽  
Heli Kirik ◽  
Annika Vilem ◽  
Laura Zani ◽  
Jan Hendrik Forth ◽  
...  

Sign in / Sign up

Export Citation Format

Share Document