Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Sep 8;10(9):564.
doi: 10.3390/vetsci10090564.

Internal Validation of the ASFV MONODOSE dtec-qPCR Kit for African Swine Fever Virus Detection under the UNE-EN ISO/IEC 17025:2005 Criteria

Gema Bru  1 , Marta Martínez-Candela  1 , Paloma Romero  1 , Aaron Navarro  1 , Antonio Martínez-Murcia  1   2
Affiliations

Internal Validation of the ASFV MONODOSE dtec-qPCR Kit for African Swine Fever Virus Detection under the UNE-EN ISO/IEC 17025:2005 Criteria

Gema Bru et al. Vet Sci. .

Abstract

African swine fever virus is considered an emerging virus that causes African swine fever, a disease characterised by high mortality and elevated transmission rates and that, as it is for most other viral diseases, cannot be treated with specific drugs. Effective and reliable detection of the virus is relevant to prevent uncontrolled contagion among boar populations and to reduce economic losses. Moreover, animal health laboratories are demanding standardisation, optimisation and quality assurance of the available diagnostic assays. In the present study, the ASFV MONODOSE dtec-qPCR kit was validated following the UNE-EN ISO/IEC 17025:2005 guidelines. Analytical validation terms include in silico and in vitro specificity, sensitivity, efficiency and reliability (repeatability/reproducibility). Diagnostic validation of the method was assessed through the analysis of a total of 181 porcine samples originating from six different matrix types doped with African swine fever virus DNA received from the European reference laboratory for African Swine Fever (INIA-CISA, Madrid, Spain): whole blood, blood serum, kidney, heart, liver and tonsil. Results agreed with those obtained from a reference detection method also based on real-time PCR, endorsed by WOAH, but the ASFV MONODOSE dtec-qPCR kit incorporates some technical innovations and improvements which may benefit end-users. This kit, available worldwide with full analytical and diagnostic validation, can recognise all known ASFV genotypes and brings additional benefits to the current qPCR technology.

Keywords: ASFV; ISO/IEC 17025:2005; diagnosis; qPCR; validation.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Quality control of the ASFV MONODOSE dtec-qPCR kit with data of six ranges of decimal dilution from 106 copies to 10 copies of the Standard Template provided in the kit, and negative control. (A) Amplification plot and (B) a representative calibration curve with stats.
Figure 2
Figure 2
Quality control of the ASFV MONODOSE dtec-qPCR kit with data of six ranges of decimal dilution from.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Alonso C., Borca M., Dixon L., Revilla Y., Rodriguez F., Escribano J.M., ICTV Report Consortium ICTV Virus Taxonomy Profile: Asfarviridae. J. Gen. Virol. 2018;99:613–614. doi: 10.1099/jgv.0.001049. - DOI - PubMed
    1. Borca M.V., Ramirez-Medina E., Silva E., Vuono E., Rai A., Pruitt S., Espinoza N., Velazquez-Salinas L., Gay C.G., Gladue D.P. ASFV-G-∆I177L as an Effective Oral Nasal Vaccine against the Eurasia Strain of Africa Swine Fever. Viruses. 2021;13:765. doi: 10.3390/v13050765. - DOI - PMC - PubMed
    1. Sánchez E.G., Pérez-Núñez D., Revilla Y. Development of vaccines against African swine fever virus. Virus Res. 2019;265:150–155. doi: 10.1016/j.virusres.2019.03.022. - DOI - PubMed
    1. Gallardo C., Casado N., Soler A., Djadjovski I., Krivko L., Madueño E., Nieto R., Perez C., Simon A., Ivanova E., et al. A multi gene-approach genotyping method identifies 24 genetic clusters within the genotype II-European African swine fever viruses circulating from 2007 to 2022. Front. Vet. Sci. 2023;10:1112850. doi: 10.3389/fvets.2023.1112850. - DOI - PMC - PubMed
    1. Quembo C., Jori F., Vosloo W., Heath L. Genetic characterization of African swine fever virus isolates from soft ticks at the wildlife/domestic interface in Mozambique and identification of a novel genotype. Transbound. Emerg. Dis. 2018;65:420–431. doi: 10.1111/tbed.12700. - DOI - PMC - PubMed