Evaluation of seven commercial African swine fever virus detection kits and three Taq polymerases on 300 well-characterized field samples

Abstract

African swine fever virus (ASFV) is a complex double stranded DNA virus, responsible for a highly infectious and fatal disease in pigs and boars and for important deterioration of animal welfare. Over the last decade, the disease spread to several European and Asian countries causing unprecedented dramatic economic losses in pig industry. In the absence of a vaccine, affected countries rely on trustful diagnostic tests and adapted testing policies to set up control programs to fight against the disease. In this study, we evaluated the sensitivity and specificity of seven commercially available ASFV real-time PCR detection kits and three Taq polymerases on 300 well-characterized wild boar samples collected in Belgium during the 2018-2019 outbreak. This study confirms that all commercial kits and two Taq polymerases are suitable for ASFV detection in diagnostic laboratories. Furthermore, the use of endogenous controls is emphasized when testing field samples harvested on carcasses in an advanced stage of decomposition, in order to avoid false negative results.

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

“…In some cases, the reduced input volume (4 µl vs 5 µl for the Kylt ASF PCR) or the lower number of PCR cycles (40 vs 45) might have led to the differences (ID Gene ASF assays). Overall, our results are in line with findings by another NRL that used seven commercially available qPCR kit for their comparison [24].…”

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

“…In some cases, the reduced input volume (4 µL vs. 5 µL for the Kylt ASF PCR) or the lower number of PCR cycles (40 vs. 45) might have led to the differences (ID Gene ASF assays). Overall, our results are in line with findings by another NRL that used seven commercially available qPCR kits for their comparison [23].…”

Towards Efficient Early Warning: Pathobiology of African Swine Fever Virus “Belgium 2018/1” in Domestic Pigs of Different Age Classes

“…For qPCR, viral nucleic acids were extracted from blood and serum using the IndiMag Pathogen Kit (Indical Bioscience, Leipzig, Germany) on the Indimag48 ® extraction platform (Indical Bioscience, Leipzig, Germany) and for tissue, the High Pure Viral Nucleic Acid Kit (Roche Applied Science, Penzberg, Germany) was used. All qPCRs were performed using the primers and probes for ASFV and endogenous gene beta-actin published by Tignon et al [ 13 ] in AgPath-ID™ One-Step RT-PCR Master mix (Applied Biosystems, Foster City, USA) as described in Schoder et al [ 14 ]. All PCRs were performed using a LC480 ® cycler (Roche, Basel, Switzerland).…”