Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Performance Characteristics of Real-time PCRs for African Swine Fever Virus Genome Detection – comparison of Twelve Kits to an OIE Recommended Method

Jutta Pikalo
ORCID logo ,
Tessa Carrau
,
Paul Deutschmann
ORCID logo ,
Melina Fischer
,
Kore Schlottau
,
Martin Beer
,
Sandra Blome
* ORCID logo
Version 1 : Received: 20 December 2021 / Approved: 21 December 2021 / Online: 21 December 2021 (09:24:26 CET)

A peer-reviewed article of this Preprint also exists.

Pikalo, J.; Carrau, T.; Deutschmann, P.; Fischer, M.; Schlottau, K.; Beer, M.; Blome, S. Performance Characteristics of Real-Time PCRs for African Swine Fever Virus Genome Detection—Comparison of Twelve Kits to an OIE-Recommended Method. Viruses 2022, 14, 220. Pikalo, J.; Carrau, T.; Deutschmann, P.; Fischer, M.; Schlottau, K.; Beer, M.; Blome, S. Performance Characteristics of Real-Time PCRs for African Swine Fever Virus Genome Detection—Comparison of Twelve Kits to an OIE-Recommended Method. Viruses 2022, 14, 220.

Abstract

African swine fever (ASF) is one of the major threats to pig production, and real-time PCR (qPCR) protocols are 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 were never 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. Concluding, all tests were fit for purpose. The test system can therefore be chosen based on compatibility and prioritization of the internal control system.

Keywords

African swine fever virus; laboratory diagnosis; commercial real-time PCR; performance; sensitivity; specificity

Subject

Biology and Life Sciences, Virology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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