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Performance Characteristics of Real-Time PCRs for African Swine Fever Virus Genome Detection—Comparison of Twelve Kits to an OIE-Recommended Method

African swine fever (ASF) is a major threat to pig production, and real-time PCR (qPCR) protocols are an integral part of ASF laboratory diagnosis. With the pandemic spread of ASF, commercial kits have risen on the market. In Germany, the kits have to go through an approval process and thus, general...

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Autores principales: Pikalo, Jutta, Carrau, Tessa, Deutschmann, Paul, Fischer, Melina, Schlottau, Kore, Beer, Martin, Blome, Sandra
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8875665/
https://www.ncbi.nlm.nih.gov/pubmed/35215814
http://dx.doi.org/10.3390/v14020220
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author Pikalo, Jutta
Carrau, Tessa
Deutschmann, Paul
Fischer, Melina
Schlottau, Kore
Beer, Martin
Blome, Sandra
author_facet Pikalo, Jutta
Carrau, Tessa
Deutschmann, Paul
Fischer, Melina
Schlottau, Kore
Beer, Martin
Blome, Sandra
author_sort Pikalo, Jutta
collection PubMed
description African swine fever (ASF) is a major threat to pig production, and real-time PCR (qPCR) protocols are an integral part of ASF laboratory diagnosis. With the pandemic spread of ASF, commercial kits have risen on the market. In Germany, the kits have to go through an approval process and thus, general validation can be assumed. However, they have never been compared to each other. In this study, 12 commercial PCR kits were compared to an OIE-recommended method. Samples representing different matrices, genome loads, and genotypes were included in a panel that was tested under diagnostic conditions. The comparison included user-friendliness, internal controls, and the time required. All qPCRs were able to detect ASFV genome in different matrices across all genotypes and disease courses. With one exception, there were no significant differences when comparing the overall mean. The overall specificity was 100% (95% CI 87.66–100), and the sensitivity was between 95% and 100% (95% CI 91.11–100). As can be expected, variability concerned samples with low genome load. To conclude, all tests were fit for purpose. The test system can therefore be chosen based on compatibility and prioritization of the internal control system.
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spelling pubmed-88756652022-02-26 Performance Characteristics of Real-Time PCRs for African Swine Fever Virus Genome Detection—Comparison of Twelve Kits to an OIE-Recommended Method Pikalo, Jutta Carrau, Tessa Deutschmann, Paul Fischer, Melina Schlottau, Kore Beer, Martin Blome, Sandra Viruses Article African swine fever (ASF) is a major threat to pig production, and real-time PCR (qPCR) protocols are an integral part of ASF laboratory diagnosis. With the pandemic spread of ASF, commercial kits have risen on the market. In Germany, the kits have to go through an approval process and thus, general validation can be assumed. However, they have never been compared to each other. In this study, 12 commercial PCR kits were compared to an OIE-recommended method. Samples representing different matrices, genome loads, and genotypes were included in a panel that was tested under diagnostic conditions. The comparison included user-friendliness, internal controls, and the time required. All qPCRs were able to detect ASFV genome in different matrices across all genotypes and disease courses. With one exception, there were no significant differences when comparing the overall mean. The overall specificity was 100% (95% CI 87.66–100), and the sensitivity was between 95% and 100% (95% CI 91.11–100). As can be expected, variability concerned samples with low genome load. To conclude, all tests were fit for purpose. The test system can therefore be chosen based on compatibility and prioritization of the internal control system. MDPI 2022-01-24 /pmc/articles/PMC8875665/ /pubmed/35215814 http://dx.doi.org/10.3390/v14020220 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Pikalo, Jutta
Carrau, Tessa
Deutschmann, Paul
Fischer, Melina
Schlottau, Kore
Beer, Martin
Blome, Sandra
Performance Characteristics of Real-Time PCRs for African Swine Fever Virus Genome Detection—Comparison of Twelve Kits to an OIE-Recommended Method
title Performance Characteristics of Real-Time PCRs for African Swine Fever Virus Genome Detection—Comparison of Twelve Kits to an OIE-Recommended Method
title_full Performance Characteristics of Real-Time PCRs for African Swine Fever Virus Genome Detection—Comparison of Twelve Kits to an OIE-Recommended Method
title_fullStr Performance Characteristics of Real-Time PCRs for African Swine Fever Virus Genome Detection—Comparison of Twelve Kits to an OIE-Recommended Method
title_full_unstemmed Performance Characteristics of Real-Time PCRs for African Swine Fever Virus Genome Detection—Comparison of Twelve Kits to an OIE-Recommended Method
title_short Performance Characteristics of Real-Time PCRs for African Swine Fever Virus Genome Detection—Comparison of Twelve Kits to an OIE-Recommended Method
title_sort performance characteristics of real-time pcrs for african swine fever virus genome detection—comparison of twelve kits to an oie-recommended method
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8875665/
https://www.ncbi.nlm.nih.gov/pubmed/35215814
http://dx.doi.org/10.3390/v14020220
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