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Sci Rep ; 7(1): 16175, 2017 11 23.
Article in English | MEDLINE | ID: mdl-29170535

ABSTRACT

Prunus necrotic ringspot virus (PNRSV) is one of the most devastating viruses to Prunus spp. In this study, we developed a diagnostic system RT-CPA-NATSC, wherein reverse transcription-cross-priming amplification (RT-CPA) is coupled with nucleic acid test strip cassette (NATSC), a vertical flow (VF) visualization, for PNRSV detection. The RT-CPA-NATSC assay targets the encoding gene of the PNRSV coat protein with a limit of detection of 72 copies per reaction and no cross-reaction with the known Prunus pathogenic viruses and viroids, demonstrating high sensitivity and specificity. The reaction is performed on 60 °C and can be completed less than 90 min with the prepared template RNA. Field sample test confirmed the reliability of RT-CPA-NATSC, indicating the potential application of this simple and rapid detection method in routine test of PNRSV.


Subject(s)
Cross-Priming/genetics , Cucumis sativus/virology , Ilarvirus/genetics , Nucleic Acid Amplification Techniques/methods , Reverse Transcription/genetics , Ilarvirus/isolation & purification , Nucleic Acids/genetics , Plant Leaves/virology
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