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Sci Rep. 2017 Nov 23;7(1):16175. doi: 10.1038/s41598-017-16536-6.

Rapid Detection of Prunus Necrotic Ringspot Virus by Reverse Transcription-cross-priming Amplification Coupled with Nucleic Acid Test Strip Cassette.

Author information

1
Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
2
Chinese Academy of Inspection and Quarantine, Beijing, 100176, China.
3
Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, China. liweimin01@caas.cn.
4
Chinese Academy of Inspection and Quarantine, Beijing, 100176, China. zhangyjpvi@yeah.net.

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.

PMID:
29170535
PMCID:
PMC5700948
DOI:
10.1038/s41598-017-16536-6
[Indexed for MEDLINE]
Free PMC Article

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