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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| Format: | Article |
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MDPI AG
2022-01-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/14/2/220 |
| _version_ | 1797476056657035264 |
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| author | Jutta Pikalo Tessa Carrau Paul Deutschmann Melina Fischer Kore Schlottau Martin Beer Sandra Blome |
| author_facet | Jutta Pikalo Tessa Carrau Paul Deutschmann Melina Fischer Kore Schlottau Martin Beer Sandra Blome |
| author_sort | Jutta Pikalo |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-09T20:52:33Z |
| format | Article |
| id | doaj.art-5f318da0fa244500b032fd4c3aa17816 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T20:52:33Z |
| publishDate | 2022-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-5f318da0fa244500b032fd4c3aa178162023-11-23T22:29:26ZengMDPI AGViruses1999-49152022-01-0114222010.3390/v14020220Performance Characteristics of Real-Time PCRs for African Swine Fever Virus Genome Detection—Comparison of Twelve Kits to an OIE-Recommended MethodJutta Pikalo0Tessa Carrau1Paul Deutschmann2Melina Fischer3Kore Schlottau4Martin Beer5Sandra Blome6Friedrich-Loeffler-Institut, Suedufer 10, 17493 Greifswald-Insel Riems, GermanyFriedrich-Loeffler-Institut, Suedufer 10, 17493 Greifswald-Insel Riems, GermanyFriedrich-Loeffler-Institut, Suedufer 10, 17493 Greifswald-Insel Riems, GermanyFriedrich-Loeffler-Institut, Suedufer 10, 17493 Greifswald-Insel Riems, GermanyFriedrich-Loeffler-Institut, Suedufer 10, 17493 Greifswald-Insel Riems, GermanyFriedrich-Loeffler-Institut, Suedufer 10, 17493 Greifswald-Insel Riems, GermanyFriedrich-Loeffler-Institut, Suedufer 10, 17493 Greifswald-Insel Riems, GermanyAfrican 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.https://www.mdpi.com/1999-4915/14/2/220African swine fever viruslaboratory diagnosiscommercial real-time PCRperformancesensitivityspecificity |
| spellingShingle | Jutta Pikalo Tessa Carrau Paul Deutschmann Melina Fischer Kore Schlottau Martin Beer Sandra Blome Performance Characteristics of Real-Time PCRs for African Swine Fever Virus Genome Detection—Comparison of Twelve Kits to an OIE-Recommended Method Viruses African swine fever virus laboratory diagnosis commercial real-time PCR performance sensitivity specificity |
| 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 | African swine fever virus laboratory diagnosis commercial real-time PCR performance sensitivity specificity |
| url | https://www.mdpi.com/1999-4915/14/2/220 |
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