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  • Title: Rapid and real-time detection of Porcine Sapelovirus by reverse transcription loop-mediated isothermal amplification assay.
    Author: Wang C, Yu D, Cui L, Hua X, Yuan C, Sun H, Liu Y.
    Journal: J Virol Methods; 2014 Jul; 203():5-8. PubMed ID: 24667302.
    Abstract:
    The present study describes the development and validation of a one-step, single-tube, and real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) detecting Porcine Sapelovirus. RT-LAMP characterized by one strand displacement reaction with the specific stem-loop structure and Bst DNA polymerase could be finished in 60 min under isothermal condition at 63 °C. RT-LAMP assay showed higher sensitivity with 10(1) copies/μL than RT-PCR for the detection of Sapelovirus. The specificity of RT-LAMP assay was validated by the absence of any cross-reaction with other closely related virus in Picornaviridae group and other common virus causing porcine diarrhea. 7 positive Sapelovirus infection out of 63 fecal samples were identified using RT-LAMP, while 5 positive samples were determined by a conventional RT-PCR. A cost-effective method for Saplovirus detection with high sensitivity and specificity was developed and evaluated.
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