Key result
A newly developed duplex fluorescent quantitative PCR assay successfully distinguished African swine fever virus genotypes I and II with a minimum detection limit of 10 copies/Rxn and high specificity.
Why the study?
Both ASFV genotypes I and II have invaded farm-raised pigs in China, creating a need for a duplex fluorescent quantitative PCR assay to distinguish between them.
A newly developed duplex fluorescent quantitative PCR assay can specifically and sensitively distinguish between ASFV genotypes I and II, providing a useful tool for epidemiological investigation and control of African swine fever.
Answers are grounded in this paper and cite their sources.
Facilitates genotype-specific ASFV detection in Chinese pigs; leaves open prospective validation before routine diagnostic use.
African swine fever (ASF) is a highly contagious hemorrhagic disease that affects domestic and wild pigs. A recent study reported that both ASF virus (ASFV) genotypes I and II have invaded farm-raised pigs in China, causing chronic infection and morbidity. To develop a duplex fluorescent quantitative PCR method to distinguish the ASFV genotypes I and II in Chinese epidemic strains, the probes and primers were designed based on the B646L sequences of genotypes I and II listed in the GenBank database. After optimizing the system, a duplex fluorescent quantitative PCR method for simultaneous detection of ASFV genotypes I and II B646L genes was successfully established. This method had no cross-reaction with Porcine circovirus type 2 (PCV2), Pseudorabies virus (PRV), or Porcine Parvovirus (PPV), indicating that it has strong specificity. The sensitivity results indicated that the minimum detection limit of ASFV genotypes I and II B646L was 10 copies/Rxn. The inter- and intra-group coefficients of variation were both <3%, indicating that the method was highly reproducible. Therefore, the established duplex fluorescent quantitative PCR assay is important for the differential detection and epidemiological investigation of ASFV.
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Cao et al. (2022) studied African swine fever virus (ASFV). Duplex fluorescent quantitative PCR assay was evaluated on Minimum detection limit and specificity. A newly developed duplex fluorescent quantitative PCR assay successfully distinguished African swine fever virus genotypes I and II with a minimum detection limit of 10 copies/Rxn and high specificity.
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