Journal article
Porcine reproductive and respiratory syndrome virus: Interlaboratory ring trial to evaluate real-time reverse transcription polymerase chain reaction detection methods : Interlaboratory ring trial to evaluate real-time reverse transcription polymerase chain reaction detection methods
Friedrich-Loeffler-Institute1
Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia2
Animal Health and Veterinary Laboratories Agency3
National Veterinary Institute4
Austrian Agency for Health and Food Safety GmbH5
National Veterinary Institute, Technical University of Denmark6
Division of Veterinary Diagnostics and Research, National Veterinary Institute, Technical University of Denmark7
Section for Veterinary Diagnostics, Division of Veterinary Diagnostics and Research, National Veterinary Institute, Technical University of Denmark8
National Veterinary Research Institute9
To compare the real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) assays used for the diagnosis of Porcine reproductive and respiratory syndrome virus (PRRSV), a Europe-wide interlaboratory ring trial was conducted. A variety of PRRSV strains including North American (NA) and European (EU) genotype isolates were analyzed by the participants.
Great differences regarding qualitative diagnostics as well as analytical sensitivity were observed between the individual RT-qPCR systems, especially when investigating strains from the EU genotype. None of the assays or commercial kits used in the ring trial could identify all different PRRSV strains with an optimal analytical and diagnostic sensitivity.
The genetic variability of the PRRSV strains, which is supposed to hinder the diagnostic of the RT-PCR because of mutations at the primer binding sites, was also confirmed by sequencing and subsequent phylogenetic analysis. In summary, a major problem in PRRSV diagnostics by RT-qPCR is false-negative results.
To achieve maximum safety in the molecular diagnosis of PRRSV, the combined usage of different assays or kits is highly recommended.
Language: | English |
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Publisher: | SAGE Publications |
Year: | 2012 |
Pages: | 855-866 |
ISSN: | 19434936 and 10406387 |
Types: | Journal article |
DOI: | 10.1177/1040638712452724 |
ORCIDs: | Hjulsager, Charlotte Kristiane |
Animals Europe Full Scientific Reports Genome, Viral Genotype Laboratories Lung Molecular Sequence Data Phylogeny Porcine Reproductive and Respiratory Syndrome Porcine respiratory and reproductive syndrome virus Real-Time Polymerase Chain Reaction Reverse Transcriptase Polymerase Chain Reaction Swine