Reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay for the visual detection of European and North American porcine reproductive and respiratory syndrome viruses

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• 作者：Ji-Young Park^a ; ^f ; Soojin Park^a ; Yu-Ri Park^a ; Dae-Young Kang^a ; ^f ; Eun-Mi Kim^a ; ^b ; Hyo-Sung Jeon^c ; Ji-Jung Kim^c ; Won-Il Kim^d ; Kyung-Tai Lee^e ; Seong-Hee Kim^f ; Kyoung-Ki Lee^f ; Choi-Kyu Park^a ; ^{parkck@knu.ac.kr" class="auth_mail" title="E-mail the corresponding author}
• 关键词：RT-LAMP ; PRRSV ; Real-time RT-PCR
• 刊名：Journal of Virological Methods
• 出版年：2016
• 出版时间：November 2016
• 年：2016
• 卷：237
• 期：Complete
• 页码：10-13
• 全文大小：681 K

文摘

A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for visual detection of European (EU) and North American (NA) porcine reproductive and respiratory syndrome viruses (PRRSVs) were established and evaluated with reference PRRSV strains and clinical samples. The assay was performed in two reaction tubes containing each set of primers specific for EU or NA-PRRSV at 58 °C for 40 min, and the results could be visually detected by the naked eye, using hydroxynaphthol blue dye. The detection limit of the assay was 1 or 0.1 TCID₅₀/0.1 mL for EU or NA PRRSV, respectively, which was comparable to that of the previously described real-time RT-PCR (qRT-PCR). The detection rate of the assay on 130 field samples was 72.3%, relatively higher than that of qRT-PCR (70.8%), and there was high overall percentage agreement between the two assays. The high specificity, sensitivity, and reliability of the RT-LAMP assay described in this study renders it useful for the rapid and differential diagnosis of EU and NA PRRSVs, even in under-equipped laboratories.