Rapid Detection of Strawberry Mild Yellow Edge Virus with a Lateral Flow Strip Reverse Transcription Recombinase Polymerase Amplification Assay

Zou, Xiaohua; Dong, Chao; Ni, Yiduo; Gao, Qing‐Hua

doi:10.1007/s00284-022-03045-7

Cited by 1 publication

(2 citation statements)

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“…Furthermore, the capability to extract DNA without complex instrumentation is a key advantage of using RPA-LFD for swift, on-site detection, and can be efficiently executed under consistent temperature conditions. For instance, Zou et al [40] established a reverse transcription recombinase polymerase amplification combined with a lateral flow strip (SMYEV-RT-RPA-LF) to diagnose strawberry mild yellow edge virus (SMYEV) in strawberries. They employed the NF-2 kit (Shenzhen Braveds Biotech Co., Ltd., Shenzhen, China) to extract RNA from strawberry leaves, using a lysis supernatant-after a five-fold serial dilution-as a direct template for RT-PCR or SMYEV-RT-RPA-LF.…”

Section: Discussionmentioning

confidence: 99%

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Real-Time Monitoring on the Chinese Giant Salamander Using RPA-LFD

Ling,

Liang,

Wang

et al. 2024

IJMS

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The Chinese giant salamander (Andrias davidianus), listed as an endangered species under “secondary protection” in China, faces significant threats due to ecological deterioration and the expansion of human activity. Extensive field investigations are crucial to ascertain the current status in the wild and to implement effective habitat protection measures to safeguard this species and support its population development. Traditional survey methods often fall short due to the elusive nature of the A. davidianus, presenting challenges that are time-consuming and generally ineffective. To overcome these obstacles, this study developed a real-time monitoring method that uses environmental DNA (eDNA) coupled with recombinase polymerase amplification and lateral flow strip (RPA-LFD). We designed five sets of species-specific primers and probes based on mitochondrial genome sequence alignments of A. davidianus and its close relatives. Our results indicated that four of these primer/probe sets accurately identified A. davidianus, distinguishing it from other tested caudata species using both extracted DNA samples and water samples from a tank housing an individual. This method enables the specific detection of A. davidianus genomic DNA at concentrations as low as 0.1 ng/mL within 50 min, without requiring extensive laboratory equipment. Applied in a field survey across four sites in Huangshan City, Anhui Province, where A. davidianus is known to be distributed, the method successfully detected the species at three of the four sites. The development of these primer/probe sets offers a practical tool for field surveying and monitoring, facilitating efforts in population recovery and resource conservation for A. davidianus.

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Section: Discussionmentioning

confidence: 99%

“…Modifications included a FAM group at the 5 ′ end, a dSpacer (oxolane, THF) at the recognition site, and a C3 spacer at the 3 ′ end. [21,40,42]. The primers were synthesized by GENEWIZ Biotechnology Co., Ltd. (Suzhou, China) and the probe by Sangong bioengineering.…”

Section: Design Of Primers and Probesmentioning

confidence: 99%