Quantitative TaqMan RT-PCR for the detection and differentiation of European and North American strains of porcine reproductive and respiratory syndrome virus.
Since two different types of porcine reproductive and respiratory syndrome virus (PRRSV), the European (EU) and the North American (US) strain, occur or coexist in European swine herds, their rapid and reliable detection and differentiation is essential for disease surveillance. A quantitative TaqMan reverse transcription-polymerase chain reaction (RT-PCR) is described for PRRSV detection and strain differentiation. Sensitivity and specificity were compared with a conventional PRRSV RT-PCR and to the detection of both PRRSV types in cell cultures and both were found to be equal or superior to the reference methods. Reproducibility was tested and proved that the assay was very reliable. Standard dilutions included in each test allowed absolute quantitation of the amount of viral RNA. The TaqMan assay described below is time-saving, easy to handle, exhibits a decreased risk of cross-contamination and is highly sensitive and specific. It is, therefore, considered to be a powerful tool for the rapid detection and differentiation of PRRSV.
Egli
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Institute of Virology and Immunoprophylaxis, Sensemattstrasse 293, CH-3147, Mittelhäusern, Switzerland.
Thür
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Liu
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L
Hofmann
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MA
D000818:Animals; D005060:Europe; D005456:Fluorescent Dyes; D009656:North America; D016133:Polymerase Chain Reaction / Q000379:methods; D019318:Porcine Reproductive and Respiratory Syndrome / Q000175:diagnosis; D019316:Porcine respiratory and reproductive syndrome virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*; D012680:Sensitivity and Specificity; D013552:Swine / Q000821:virology*; D019914:Taq Polymerase
D005456:Fluorescent Dyes; D012367:RNA, Viral; D019914:Taq Polymerase