ArticleApplied and environmental microbiology2014
Primer modification improves rapid and sensitive in vitro and field-deployable assays for detection of high plains virus variants.
Article in Applied and environmental microbiology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 37 citations in OpenAlex.
- Advances in Isothermal Amplification for the Diagnosis of Tuberculosis.Journal of clinical laboratory analysis · 2025Review
- Isothermal Amplification Techniques: An Emerging Tool for On-Site Detection of Phytopathogens in Field Conditions.Methods in molecular biology (Clifton, N.J.) · 2025Review
- Development of a highly sensitive TaqMan method based on multi-probe strategy: its application in ASFV detection.Biology methods & protocols · 2024Article
- In-silico prediction of RT-qPCR-high resolution melting for broad detection of emaraviruses.PloS one · 2023Article
- Pest categorisation of High Plains wheat mosaic virus.EFSA journal. European Food Safety Authority · 2022Article
- Development and Validation of a New TaqMan Real-Time PCR for the Detection ofMicroorganisms · 2022Article
- Loop-Mediated Isothermal Amplification for Detection of Plant Pathogens in Wheat (Frontiers in plant science · 2022Review
- The Potential Use of Isothermal Amplification Assays for In-Field Diagnostics of Plant Pathogens.Plants (Basel, Switzerland) · 2021Review
- High Plains wheat mosaic virus: An enigmatic disease of wheat and corn causing the High Plains disease.Molecular plant pathology · 2021Article
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- Genome-informed diagnostics for specific and rapid detection of Pectobacterium species using recombinase polymerase amplification coupled with a lateral flow device.Scientific reports · 2018Article
- Helicase-dependent isothermal amplification: a novel tool in the development of molecular-based analytical systems for rapid pathogen detection.Analytical and bioanalytical chemistry · 2018Review
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Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A high consequence pathogen, High plains virus (HPV) causes considerable damage to wheat if the crop is infected during early stages of development. Methods for the early, accurate, and sensitive detection of HPV in plant tissues are needed for the management of disease outbreaks and reservoir hosts. In this study, the effectiveness of five methods-real-time SYBR green and TaqMan reverse transcription-quantitative PCR (RT-qPCR), endpoint RT-PCR, RT-helicase dependent amplification (RT-HDA) and the Razor Ex BioDetection System (Razor Ex)-for the broad-range detection of HPV variants was evaluated. Specific PCR primer sets and probes were designed to target the HPV nucleoprotein gene. Primer set HPV6F and HPV4R, which amplifies a product of 96 bp, was validated in silico against published sequences and in vitro against an inclusivity panel of infected plant samples and an exclusivity panel of near-neighbor viruses. The primers were modified by adding a customized 22 nucleotide long tail at the 5' terminus, raising the primers' melting temperature (Tm; ca. 10°C) to make them compatible with RT-HDA (required optimal Tm = 68°C), in which the use of primers lacking such tails gave no amplification. All of the methods allowed the detection of as little as 1 fg of either plasmid DNA carrying the target gene sequence or of infected plant samples. The described in vitro and in-field assays are accurate, rapid, sensitive, and useful for pathogen detection and disease diagnosis, microbial quantification, and certification and breeding programs, as well as for biosecurity and microbial forensics applications.
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