Abstract
BackgroundPorcine circovirus type 3 (PCV3) is an emerging circovirus species, that has been reported in major pig-raising countries including the United States, China, South Korea, Brazil, Spain, and Poland.ResultsA real-time loop-mediated isothermal amplification (LAMP) assay was developed for rapid detection of porcine circovirus 3 (PCV3). The method had a detection limit of 1 × 101 copies/μL with no cross-reactions with classical swine fever virus (CSFV) C strain, foot-and-mouth disease virus (FMDV), porcine circovirus 2 (PCV2) LG vaccine strain, porcine epidemic diarrhoea virus (PEDV), porcine respiratory and reproductive syndrome virus (PRRSV), or pseudorabies virus (PRV). The PCV3 positive detection rate of 203 clinical samples for the real-time LAMP assay was 89.66% (182/203).ConclusionsThe real-time LAMP assay is highly sensitive, and specific for use in epidemiological investigations of PCV3.
Highlights
Porcine circovirus type 3 (PCV3) is an emerging circovirus species, that has been reported in major pig-raising countries including the United States, China, South Korea, Brazil, Spain, and Poland
Sensitivity and specificity of the real-time loop-mediated isothermal amplification (LAMP) assay for rapid detection of porcine circovirus 3 (PCV3) Ten-fold serial dilutions of plasmid DNA were used as templates in the assay
The detection limit of the assay was 1 × 101 copies/μL per reaction (Fig. 1). These results indicated the real-time LAMP assay could be used as a sensitive diagnostic test for PCV3
Summary
Porcine circovirus type 3 (PCV3) is an emerging circovirus species, that has been reported in major pig-raising countries including the United States, China, South Korea, Brazil, Spain, and Poland. In June 2015, an outbreak of porcine dermatitis and nephropathy syndrome (PDNS) was reported in a commercial pig farm in North Carolina, United States. Given that the virus may be highly prevalent globally, an accurate laboratory diagnosic for rapid confirmation of PCV3 infection is needed. Conventional loop-mediated isothermal amplification (LAMP) assays and PCR-based diagnostic assays for PCV3 have been developed [8,9,10]. Real-time LAMP is a constant temperature amplification method carried out at 60–65 °C, for which only a simple water bath is required. Real-time LAMP eliminates reverse transcription steps as well as PCR instrument cooling time, which shortens the amplification time.
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