Development and validation of a fast quantitative real-time PCR assay for the detection of African swine fever virus.
African swine fever virus
diagnostics
fast
quantitative
real-time PCR
Journal
Frontiers in veterinary science
ISSN: 2297-1769
Titre abrégé: Front Vet Sci
Pays: Switzerland
ID NLM: 101666658
Informations de publication
Date de publication:
2022
2022
Historique:
received:
06
09
2022
accepted:
04
10
2022
entrez:
23
1
2023
pubmed:
24
1
2023
medline:
24
1
2023
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 36686190
doi: 10.3389/fvets.2022.1037728
pmc: PMC9845278
doi:
Types de publication
Journal Article
Langues
eng
Pagination
1037728Informations de copyright
Copyright © 2023 Hwang, Choi, Song, Frant and Kim.
Déclaration de conflit d'intérêts
Authors HJH, YSC, and KS were employed by Ahram Biosystems, Inc. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
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