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Ring tests to evaluate the performance of Porcine circovirus-2 (PCV-2) polymerase chain reaction (PCR) assays used in North American diagnostic laboratories.

ABSTRACT: Two laboratory studies involving 11 laboratories were undertaken to assess the performance of North American Porcine circovirus-2 (PCV-2) polymerase chain reaction (PCR) assays. Laboratories received identical submissions containing randomly coded positive and negative control samples, and serially diluted PCV-2-spiked samples. In study 1 and 2, respectively, spiked samples contained measured amounts of PCV-2 virus or DNA. All but 1 assay detected DNA in the most concentrated spiked sample. There were no statistical differences in the proportion of positive or negative samples reported by quantitative (n = 7) versus non-quantitative (n = 6) assays. Across both studies, the false positive rate was 17% (4 out of 23), and 17% (2 out of 12) of assays cross-reacted with PCV-1. The most sensitive assay detected PCV-2 DNA levels about 100 000 times lower the least sensitive assay. This study demonstrated that the PCR assays available in North American diagnostic labs vary considerably in their detection limits and quantification.

SUBMITTER: Harding JC

PROVIDER: S-EPMC2613600 | biostudies-literature | 2009 Jan

REPOSITORIES: biostudies-literature

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Ring tests to evaluate the performance of Porcine circovirus-2 (PCV-2) polymerase chain reaction (PCR) assays used in North American diagnostic laboratories.

Harding John C S JC Baker Crissie C Rhodes Carrie C McIntosh Kathleen A KA Bonneau Martin M

Canadian journal of veterinary research = Revue canadienne de recherche veterinaire 20090101 1

Two laboratory studies involving 11 laboratories were undertaken to assess the performance of North American Porcine circovirus-2 (PCV-2) polymerase chain reaction (PCR) assays. Laboratories received identical submissions containing randomly coded positive and negative control samples, and serially diluted PCV-2-spiked samples. In study 1 and 2, respectively, spiked samples contained measured amounts of PCV-2 virus or DNA. All but 1 assay detected DNA in the most concentrated spiked sample. Ther ...[more]

PMID: 19337389

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Project description:Porcine circovirus 2 (PCV-2) is one of the most widespread viral infections of swine, causing a remarkable economic impact because of direct losses and indirect costs for its control. As other ssDNA viruses, PCV-2 is characterized by a high evolutionary rate, leading to the emergence of a plethora of variants with different biological and epidemiological features. Over time, several attempts have been made to organize PCV-2 genetic heterogeneity in recognized genotypes. This categorization has clearly simplified the epidemiological investigations, allowing to identify different spatial and temporal patterns among genotypes. Additionally, variable virulence and vaccine effectiveness have also been hypothesized. However, the rapid increase in sequencing activity, coupled with the per se high viral variability, has challenged the previously established nomenclature, leading to the definition of several study-specific genotypes and hindering the capability of performing comparable epidemiological studies. Based on these premises, an updated classification scheme is herein reported. Recognizing the impossibility of defining a clear inter-cluster p-distance cut-off, the present study proposes a phylogeny-grounded genotype definition based on three criteria: maximum intra-genotype p-distance of 13% (calculated on the ORF2 gene), bootstrap support at the corresponding internal node higher than 70% and at least 15 available sequences. This scheme allowed defining 8 genotypes (PCV-2a to PCV-2h), which six of those had been previously proposed. To minimize the inconvenience of implementing a new classification, the most common names already adopted have been maintained when possible. The analysis of sequence-associated metadata highlighted a highly unbalanced sequencing activity in terms of geographical, host and temporal distribution. The PCV-2 molecular epidemiology scenario appears therefore characterized by a severe bias that could lead to spurious associations between genetic and epidemiological/biological viral features. While the suggested classification can establish a "common language" for future studies, further efforts should be paid to achieve a more homogeneous and informative representation of the PCV-2 global scenario.

Dataset Information

Ring tests to evaluate the performance of Porcine circovirus-2 (PCV-2) polymerase chain reaction (PCR) assays used in North American diagnostic laboratories.

Publications

Ring tests to evaluate the performance of Porcine circovirus-2 (PCV-2) polymerase chain reaction (PCR) assays used in North American diagnostic laboratories.

Similar Datasets

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