Development of a loop‐mediated isothermal amplification assay for rapid and simple detection of Erysipelothrix rhusiopathiae. (11th December 2013)

• Record Type: Journal Article
• Authors: Yamazaki, Y.; Oba, E.; Kashiwagi, N.; Sugita, K.; Shiiba, K.; Baba, Y.; Shimoji, Y.; Yamazaki, W.
• Is Part Of: Letters in applied microbiology. Volume 58:Number 4(2014:Apr.)
• Page Start: 362
• Page End: 369
• Publication Date: 2013-12-11
• Subjects: Microbiology -- Periodicals; 660.62
• DOI: 10.1111/lam.12198
• Languages: English
• ISSNs: 0266-8254

Abstract

<abstract abstract-type="main" id="lam12198-abs-0001"> <title> <x xml:space="preserve">Abstract</x> </title> <sec id="lam12198-sec-0102" sec-type="section"> <title>Abstract</title> <p> <italic>Erysipelothrix rhusiopathiae</italic> is a causative agent of swine erysipelas. We developed a novel and highly specific loop‐mediated isothermal amplification (LAMP) assay for sensitive and rapid detection of <italic>E. rhusiopathiae</italic>. The LAMP assay correctly detected 39 <italic>E. rhusiopathiae</italic> strains. No LAMP products were detected from 14 non‐<italic>rhusiopathiae Erysipelothrix</italic> and 16 non‐<italic>Erysipelothrix</italic> strains, including <italic>E. tonsillarum</italic> serovar 10 strains, which are difficult to be discriminated from <italic>E. rhusiopathiae</italic> strains. These results were consistent with those obtained by a conventional <italic>E. rhusiopathiae</italic>‐specific PCR assay. Starting with DNA extraction from a single colony, the gel‐based PCR assay took 4 h to provide a result, but the LAMP assay was faster, requiring only 37–80 min. The conventional culture test required more than 3–4 days to isolate and identify <italic>E. rhusiopathiae</italic> in the enrichment cultures. In contrast, the LAMP assay required less than 22 h from the beginning of the enrichment culture to final determination. These results suggest that the LAMP assay is useful as an adjunct to facilitate early diagnosis of swine erysipelas.</p> </sec> <sec id="lam12198-sec-0112" sec-type="section"> <title>Significance and Impact of the Study</title> <p>This is the first report of a loop‐mediated isothermal amplification (LAMP) assay for simple and cost‐effective detection of <italic>E. rhusiopathiae</italic> from swine samples. The LAMP assay provided more rapid detection of the bacterium than conventional PCR and biochemical‐based assays, and it may potentially facilitate surveillance and early diagnosis of swine erysipelas in the field.</p> </sec> </abstract>