Rapid and sensitive detection of porcine torovirus by a reverse transcription loop-mediated isothermal amplification assay (RT-LAMP)

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• 作者：Xiaowan Liu^a ; ^{abtclxw@126.com" class="auth_mail" title="E-mail the corresponding author} ; Yuancheng Zhou^a ; ^{god098@163.com" class="auth_mail" title="E-mail the corresponding author} ; Fan Yang^a ; ^{abtcyf@126.com" class="auth_mail" title="E-mail the corresponding author} ; Pengjuan Liu^a ; ^{1210695671@qq.com" class="auth_mail" title="E-mail the corresponding author} ; Yuhan Cai^a ; ^{429197524@qq.com" class="auth_mail" title="E-mail the corresponding author} ; Jianbo Huang^a ; ^{514570512@qq.com" class="auth_mail" title="E-mail the corresponding author} ; Ling Zhu^a ; ^b ; ^{abtczl72@126.com" class="auth_mail" title="E-mail the corresponding author} ; Zhiwen Xu^a ; ^b ; ^{abtcxzw@126.com" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Rapid detection ; Porcine torovirus ; Reverse transcription loop-mediated isothermal amplification assay
• 刊名：Journal of Virological Methods
• 出版年：2016
• 出版时间：February 2016
• 年：2016
• 卷：228
• 期：Complete
• 页码：103-107
• 全文大小：775 K

文摘

Porcine torovirus (PToV) is associated with swine gastroenteritis, but its pathogenesis is uncertain because there is limited information regarding PToV due to its difficulty to adapt in vitro. This study has developed a rapid one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for the detection of PToV. A set of four primers specific to six regions within the PToV's highly conserved fragment of the M gene was designed for use with the RT-LAMP assay. The RT-LAMP assay was sensitive with a detection limit of 1 × 10¹ copies/μL, which was 100-fold higher than reverse-transcription PCR. No cross-reaction was observed with other similar viruses. A total of 175 clinical specimens were collected from the Sichuan province, and PToV was detected by the established RT-LAMP assay with a positive rate of 39.2% (69/175). This study developed the first rapid, sensitive, simple, cost-effective and accurate method for the detection of PToV. The results show that the RT-LAMP assay is highly feasible in clinical settings.