Development of a Real-Time Recombinase Polymerase Amplification Assay for the Rapid Detection of African Swine Fever Virus Genotype I and II

African swine fever (ASF) is a contagious viral disease in pigs and wild boars which poses a major threat to the pig industry. Rapid and accurate diagnosis is necessary to control ASF. Hence, we developed a rapid diagnostic method using a recombinase polymerase amplification (RPA) assay targeting th...

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Bibliographic Details
Published in:Pathogens (Basel) 2022-04, Vol.11 (4), p.439
Main Authors: Ilya, Titov, Monoldorova, Sezim, Kang, Shin-Seok, Yun, Seungri, Byeon, Hyeon-Seop, Mariia, Nefedeva, Jeon, Bo-Young
Format: Article
Language:English
Subjects:
African swine fever
African swine fever virus (ASFV)
Amplification
Asfarviridae
Assaying
Confidence intervals
CP204 gene
Disease prevention
Epidemics
Fever
Genomes
Genotype & phenotype
genotype I
genotype II
Genotypes
Hogs
Infections
Methods
Mutation
Polymerase chain reaction
Real time
Recombinase
recombinase polymerase amplification (RPA)
Swine
Thermal cycling
Viral diseases
Viruses
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Summary:African swine fever (ASF) is a contagious viral disease in pigs and wild boars which poses a major threat to the pig industry. Rapid and accurate diagnosis is necessary to control ASF. Hence, we developed a rapid diagnostic method using a recombinase polymerase amplification (RPA) assay targeting the conserved sequences of CP204L (p30) thatcan rapidly detect ASF virus (ASFV) genotype strains I and II. The lower detection limit of the real-time RPA assay was 5 × 101 copies per reaction. The real-time RPA assay effectively detected ASFV isolates and clinical specimens belonging to ASFV genotypes I and II. The sensitivity and specificity of the assay were 96.8% (95% confidence interval (CI): 83.3−99.9) and 100% (95% CI: 88.4−100.0), respectively. The agreement between the real-time RPA assay and a reference commercial real-time quantitative polymerase chain reaction (qPCR) was 100%. The real-time RPA assay had a detection time of 6.0 min (95% CI: 5.7−6.2), which was significantly shorter than that of qPCR (49 min; 95% CI: 47.4−50.6; p < 0.001). Thus, the developed real-time RPA assay is a rapid and accurate diagnostic tool for detecting ASFV genotypes I and II.
ISSN:2076-0817
2076-0817
DOI:10.3390/pathogens11040439