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Development and validation of a fast quantitative real-time PCR assay for the detection of African swine fever virus
African swine fever virus (ASFV) is a double-stranded DNA virus that causes African swine fever (ASF), a lethal hemorrhagic fever that is highly contagious among domestic pigs and wild boars. Due to the high mortality rates and highly contagious nature of the ASF, it is important to develop a fast d...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2023
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9845278/ https://www.ncbi.nlm.nih.gov/pubmed/36686190 http://dx.doi.org/10.3389/fvets.2022.1037728 |
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author | Hwang, Hyun Jin Choi, Yun Seong Song, Kyungyoung Frant, Maciej Kim, Jeong Hee |
author_facet | Hwang, Hyun Jin Choi, Yun Seong Song, Kyungyoung Frant, Maciej Kim, Jeong Hee |
author_sort | Hwang, Hyun Jin |
collection | PubMed |
description | African swine fever virus (ASFV) is a double-stranded DNA virus that causes African swine fever (ASF), a lethal hemorrhagic fever that is highly contagious among domestic pigs and wild boars. Due to the high mortality rates and highly contagious nature of the ASF, it is important to develop a fast detection method for ASFV with high sensitivity and specificity to take an immediate action to stop wide spread of the virulent disease. Therefore, a fast and quantitative molecular detection method of ASFV is presented in this study. A total of 24 genotypes of ASFV have been identified based on nucleic acid sequences of the major capsid protein p72. The primers and probe of the present assay was designed to detect all of the p72-based genotypes of ASFV. The turnaround time for PCR detection was within 50 min which is at least about two-times faster compared to other PCR assays. Limit of detection (LoD) was 6.91 genomic copies/reaction for the most virulent genotype II. LoD values for other genotypes were within 10–20 copies/reaction. Cross-reactivity of the assay was validated using a panel of pathogens related to swine disease, and no cross-reactivity was observed. Positive and negative clinical samples (50 samples each) obtained from sick and healthy animals, were used to validate the assay. The results showed that 100% agreement for both positive and negative samples. In summary, the assay described in this study offers the advantage of rapid detection of all genotypes of ASFV with high sensitivity and specificity. The assay is a valuable tool both in clinical and laboratory uses for sensitive and fast detection of ASFV. |
format | Online Article Text |
id | pubmed-9845278 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-98452782023-01-19 Development and validation of a fast quantitative real-time PCR assay for the detection of African swine fever virus Hwang, Hyun Jin Choi, Yun Seong Song, Kyungyoung Frant, Maciej Kim, Jeong Hee Front Vet Sci Veterinary Science African swine fever virus (ASFV) is a double-stranded DNA virus that causes African swine fever (ASF), a lethal hemorrhagic fever that is highly contagious among domestic pigs and wild boars. Due to the high mortality rates and highly contagious nature of the ASF, it is important to develop a fast detection method for ASFV with high sensitivity and specificity to take an immediate action to stop wide spread of the virulent disease. Therefore, a fast and quantitative molecular detection method of ASFV is presented in this study. A total of 24 genotypes of ASFV have been identified based on nucleic acid sequences of the major capsid protein p72. The primers and probe of the present assay was designed to detect all of the p72-based genotypes of ASFV. The turnaround time for PCR detection was within 50 min which is at least about two-times faster compared to other PCR assays. Limit of detection (LoD) was 6.91 genomic copies/reaction for the most virulent genotype II. LoD values for other genotypes were within 10–20 copies/reaction. Cross-reactivity of the assay was validated using a panel of pathogens related to swine disease, and no cross-reactivity was observed. Positive and negative clinical samples (50 samples each) obtained from sick and healthy animals, were used to validate the assay. The results showed that 100% agreement for both positive and negative samples. In summary, the assay described in this study offers the advantage of rapid detection of all genotypes of ASFV with high sensitivity and specificity. The assay is a valuable tool both in clinical and laboratory uses for sensitive and fast detection of ASFV. Frontiers Media S.A. 2023-01-04 /pmc/articles/PMC9845278/ /pubmed/36686190 http://dx.doi.org/10.3389/fvets.2022.1037728 Text en Copyright © 2023 Hwang, Choi, Song, Frant and Kim. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Veterinary Science Hwang, Hyun Jin Choi, Yun Seong Song, Kyungyoung Frant, Maciej Kim, Jeong Hee Development and validation of a fast quantitative real-time PCR assay for the detection of African swine fever virus |
title | Development and validation of a fast quantitative real-time PCR assay for the detection of African swine fever virus |
title_full | Development and validation of a fast quantitative real-time PCR assay for the detection of African swine fever virus |
title_fullStr | Development and validation of a fast quantitative real-time PCR assay for the detection of African swine fever virus |
title_full_unstemmed | Development and validation of a fast quantitative real-time PCR assay for the detection of African swine fever virus |
title_short | Development and validation of a fast quantitative real-time PCR assay for the detection of African swine fever virus |
title_sort | development and validation of a fast quantitative real-time pcr assay for the detection of african swine fever virus |
topic | Veterinary Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9845278/ https://www.ncbi.nlm.nih.gov/pubmed/36686190 http://dx.doi.org/10.3389/fvets.2022.1037728 |
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