Next Article in Journal
Phelan McDermid Syndrome: Multiple Sclerosis as a Rare but Treatable Cause for Regression—A Case Report
Previous Article in Journal
Targeting Epigenetic Regulators for Endometrial Cancer Therapy: Its Molecular Biology and Potential Clinical Applications
Previous Article in Special Issue
Optimized qRT-PCR Approach for the Detection of Intra- and Extra-Cellular SARS-CoV-2 RNAs
Article

Swift and Reliable “Easy Lab” Methods for the Sensitive Molecular Detection of African Swine Fever Virus

Friedrich-Loeffler-Institut, Institute of Diagnostic Virology, 17493 Greifswald-Insel Riems, Germany
*
Author to whom correspondence should be addressed.
Academic Editor: Ilka Engelmann
Int. J. Mol. Sci. 2021, 22(5), 2307; https://doi.org/10.3390/ijms22052307
Received: 27 January 2021 / Revised: 16 February 2021 / Accepted: 21 February 2021 / Published: 25 February 2021
(This article belongs to the Special Issue Molecular Detection and Typing of Viruses)
African swine fever (ASF) is a contagious viral hemorrhagic disease of domestic pigs and wild boars. The disease is notifiable to the World Organisation for Animal Health (OIE) and is responsible for high mortality and serious economic losses. PCR and real-time PCR (qPCR) are the OIE-recommended standard methods for the direct detection of African swine fever virus (ASFV) DNA. The aim of our work was the simplification and standardization of the molecular diagnostic workflow in the lab. For validation of this “easy lab” workflow, different sample materials from animal trials were collected and analyzed (EDTA blood, serum, oral swabs, chewing ropes, and tissue samples) to identify the optimal sample material for diagnostics in live animals. Based on our data, the EDTA blood samples or bloody tissue samples represent the best specimens for ASFV detection in the early and late phases of infection. The application of prefilled ready-to-use reagents for nucleic acid extraction or the use of a Tissue Lysis Reagent (TLR) delivers simple and reliable alternatives for the release of the ASFV nucleic acids. For the qPCR detection of ASFV, different published and commercial kits were compared. Here, a lyophilized commercial kit shows the best results mainly based on the increased template input. The good results of the “easy lab” strategy could be confirmed by the ASFV detection in field samples from wild boars collected from the 2020 ASFV outbreak in Germany. Appropriate internal control systems for extraction and PCR are key features of the “easy lab” concept and reduce the risk of false-negative and false-positive results. In addition, the use of easy-to-handle machines and software reduces training efforts and the misinterpretation of results. The PCR diagnostics based on the “easy lab” strategy can realize a high sensitivity and specificity comparable to the standard PCR methods and should be especially usable for labs with limited experiences and resources. View Full-Text
Keywords: African swine fever virus; DNA extraction; real-time PCR; easy lab African swine fever virus; DNA extraction; real-time PCR; easy lab
Show Figures

Figure 1

MDPI and ACS Style

Elnagar, A.; Pikalo, J.; Beer, M.; Blome, S.; Hoffmann, B. Swift and Reliable “Easy Lab” Methods for the Sensitive Molecular Detection of African Swine Fever Virus. Int. J. Mol. Sci. 2021, 22, 2307. https://doi.org/10.3390/ijms22052307

AMA Style

Elnagar A, Pikalo J, Beer M, Blome S, Hoffmann B. Swift and Reliable “Easy Lab” Methods for the Sensitive Molecular Detection of African Swine Fever Virus. International Journal of Molecular Sciences. 2021; 22(5):2307. https://doi.org/10.3390/ijms22052307

Chicago/Turabian Style

Elnagar, Ahmed, Jutta Pikalo, Martin Beer, Sandra Blome, and Bernd Hoffmann. 2021. "Swift and Reliable “Easy Lab” Methods for the Sensitive Molecular Detection of African Swine Fever Virus" International Journal of Molecular Sciences 22, no. 5: 2307. https://doi.org/10.3390/ijms22052307

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop