Reverse transcription loop-mediated isothermal amplification for the detection of Porcine reproductive and respiratory syndrome virus

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Abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) is an important pathogen of swine. The objective of the current study is to investigate the feasibility of using reverse transcription loop-mediated isothermal amplification (RT-LAMP) for the detection of PRRSV. The RT-LAMP is a recently described DNA amplification technique reported to be simple, inexpensive, fast, and accurate. The RT-LAMP reaction was set up using 2 sets of primers that were designed to detect North American and European strains of PRRSV and performed successfully in a simple heat block. The specificity of the amplified product was demonstrated by restriction analysis. The RT-LAMP was able to detect 5 different PRRSV isolates.However, the limit of detection ranged between 102 and 104 50% tissue culture infective dose/ml. The RT-LAMP was further evaluated using serum samples from animals of known infection status. The ability of RT-LAMP to detect PRRSV in serum from acutely infected animals was evaluated with 114 serum samples from 18 experimentally inoculated boars. Fortynine of these samples tested positive by RT-LAMP, while 94 were positive by reverse transcription polymerase chain reaction (RT-PCR). The diagnostic specificity, evaluated with 100 known negative serum samples, was estimated as 99%. The feasibility of RT-LAMP to detect PRRSV was demonstrated in the current study. The RTLAMP reaction could be performed in just 1 hr with a simple and inexpensive heat block.However, the sensitivity of this technique was significantly lower than that of RT-PCR.

Original languageEnglish (US)
Pages (from-to)350-354
Number of pages5
JournalJournal of Veterinary Diagnostic Investigation
Volume21
Issue number3
DOIs
StatePublished - May 1 2009

Keywords

  • Diagnostic test
  • Loop-mediated isothermal amplification
  • Polymerase chain reaction
  • Porcine reproductive and respiratory syndrome virus

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