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Multiplex polymerase chain reaction for the detection and differentiation of 4 porcine circovirus 2 genotypes (PCV-2a, -2b, -2d, and -2e) in clinical samples.

ABSTRACT: The objective of this study was to develop multiplex polymerase chain reaction (PCR) for the simultaneous detection of porcine circovirus 2 (PCV-2) and differentiation among 4 PCV-2 genotypes (2a, 2b, 2d, and 2e) in collected clinical lymph node samples. The multiplex PCR detected each of 4 PCV-2 genotypes (2a, 2b, 2d, and 2e) to a dilution of 2 × 10¹ copies/μL. PCV-2a, PCV-2b, PCV-2d, and PCV-2e were propagated in tissues prior to DNA extraction for use in multiplex PCR for the simultaneous detection and differentiation of 4 PCV-2 genotypes. The designed multiplex PCR effectively detected and differentiated various combinations of multiple infection, such as PCV-2a+2b, PCV-2a+2d, PCV-2b+2d, PCV-2a+2e, and PCV-2a+2b+2d, in clinical lymph node samples. The results of this study demonstrated that multiplex PCR testing of clinical samples developed herein was able to simultaneously detect and differentiate among the 4 PCV-2 genotypes (PCV-2a, 2b, 2d, and 2e).

SUBMITTER: Park KH

PROVIDER: S-EPMC8978285 | biostudies-literature | 2022 Apr

REPOSITORIES: biostudies-literature

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Multiplex polymerase chain reaction for the detection and differentiation of 4 porcine circovirus 2 genotypes (PCV-2a, -2b, -2d, and -2e) in clinical samples.

Park Kee Hwan KH Cho Hyejean H Oh Taehwan T Yang Siyeon S Chae Chanhee C

Canadian journal of veterinary research = Revue canadienne de recherche veterinaire 20220401 2

The objective of this study was to develop multiplex polymerase chain reaction (PCR) for the simultaneous detection of porcine circovirus 2 (PCV-2) and differentiation among 4 PCV-2 genotypes (2a, 2b, 2d, and 2e) in collected clinical lymph node samples. The multiplex PCR detected each of 4 PCV-2 genotypes (2a, 2b, 2d, and 2e) to a dilution of 2 × 10<sup>1</sup> copies/μL. PCV-2a, PCV-2b, PCV-2d, and PCV-2e were propagated in tissues prior to DNA extraction for use in multiplex PCR for the simul ...[more]

PMID: 35388234

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Project description:IntroductionPorcine circovirus 2 (PCV-2) is a key pathogen for the swine industry at a global level. Nine genotypes, differing in epidemiology and potentially virulence, emerged over time, with PCV-2a, -2b, and -2d being the most widespread and clinically relevant. Conversely, the distribution of minor genotypes appears geographically and temporally restricted, suggesting lower virulence and different epidemiological drivers. In 2022, PCV-2e, the most genetically and phenotypically divergent genotype, was identified in multiple rural farms in North-eastern Italy. Since rural pigs often have access to outdoor environment, the introduction from wild boars was investigated.MethodsThrough a molecular and spatial approach, this study investigated the epidemiology and genetic diversity of PCV-2 in 122 wild boars across different provinces of North-eastern Italy.ResultsMolecular analysis revealed a high PCV-2 frequency (81.1%, 99/122), and classified the majority of strains as PCV-2d (96.3%, 78/81), with sporadic occurrences of PCV-2a (1.2%, 1/81) and PCV-2b (2.5%, 2/81) genotypes. A viral flow directed primarily from domestic pigs to wild boars was estimated by phylogenetic and phylodynamic analyses.DiscussionThese findings attested that the genotype replacement so far described only in the Italian domestic swine sector occurred also in wild boars. and suggested that the current heterogeneity of PCV-2d strains in Italian wild boars likely depends more on different introduction events from the domestic population rather than the presence of independent evolutionary pressures. While this might suggest PCV-2 circulation in wild boars having a marginal impact in the industrial sector, the sharing of PCV-2d strains across distinct wild populations, in absence of a consistent geographical pattern, suggests a complex interplay between domestic and wild pig populations, emphasizing the importance of improved biosecurity measures to mitigate the risk of pathogen transmission.

Dataset Information

Multiplex polymerase chain reaction for the detection and differentiation of 4 porcine circovirus 2 genotypes (PCV-2a, -2b, -2d, and -2e) in clinical samples.

Publications

Multiplex polymerase chain reaction for the detection and differentiation of 4 porcine circovirus 2 genotypes (PCV-2a, -2b, -2d, and -2e) in clinical samples.

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