A semi-automated and highly sensitive streptavidin magnetic capture-hybridization RT-PCR assay: Application to genus-wide or species-specific detection of several viruses of ornamental bulb crops

Miglino, R.; Jodlowska, A.; Pappu, H. R.; Schadewijk, Ton R. van

doi:10.1016/j.jviromet.2007.06.014

Cited by 7 publications

(5 citation statements)

References 28 publications

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“…PCR tests have been used to detect TRV in vector nematodes, also using RNA2specific tests when defined TRV isolates are under study . Increased sensitivity has been achieved using capture hybridization prior to reverse transcriptase-polymerase chain reaction (RT-PCR) (Miglino et al, 2007) or modifications to sample preparation (Martin et al, 2009).…”

Section: Trv As a Potato Pathogenmentioning

confidence: 99%

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Tobraviruses—plant pathogens and tools for biotechnology

MacFarlane¹

2010

Molecular Plant Pathology

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The tobraviruses, Tobacco rattle virus (TRV), Pea early-browning virus (PEBV) and Pepper ringspot virus (PepRSV), are positive-strand RNA viruses with rod-shaped virus particles that are transmitted between plants by trichodorid nematodes. As a group, these viruses infect many plant species, with TRV having the widest host range. Recent studies have begun to dissect the interaction of TRV with potato, currently the most commercially important crop disease caused by any of the tobraviruses. As well as being successful plant pathogens, these viruses have become widely used as vectors for expression in plants of nonviral proteins or, more frequently, as initiators of virus-induced gene silencing (VIGS). Precisely why tobraviruses should be so effective as VIGS vectors is not known; however, molecular studies of the mode of action of the tobravirus silencing suppressor protein are shedding some light on this process.

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Section: Trv As a Potato Pathogenmentioning

confidence: 99%

“…, 2006). Increased sensitivity has been achieved using capture hybridization prior to reverse transcriptase‐polymerase chain reaction (RT‐PCR) (Miglino et al. , 2007) or modifications to sample preparation (Martin et al.…”

Section: Trv As a Potato Pathogenmentioning

confidence: 99%

Tobraviruses—plant pathogens and tools for biotechnology

MacFarlane¹

2010

Molecular Plant Pathology

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“…MCH is sensitive and specific and relies on a nucleic acid probe to bind to and concentrate target sequences. MCH has been adapted for detection of bacteria, fungi, and viruses from a wide variety of research fields (1,16,24). IMS relies on antibodies for selective targeting of the pathogen for use in real-time PCR.…”

Section: Discussionmentioning

confidence: 99%

Development of a Magnetic Capture Hybridization Real-Time PCR Assay for Detection of TumorigenicAgrobacterium vitisin Grapevines

Johnson¹,

Zheng²,

Kaewnum³

et al. 2013

Phytopathology®

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Agrobacterium vitis, the causal agent of grape crown gall, can have severe economic effects on grape production. The bacterium survives systemically in vines and, therefore, is disseminated in propagation material. We developed an assay for use in indexing programs that is efficient and sensitive for detecting A. vitis in grape tissue. Initially, real-time polymerase chain reaction (PCR) primers specific for diverse tumorigenic strains of A. vitis were developed using the virD2 gene sequence. To overcome the effects of PCR inhibitors present in plant tissue, DNA extraction methods that included magnetic capture hybridization (MCH), immunomagnetic separation (IMS), and extraction with the Mo Bio Powerfood kit were compared. The assays incorporating MCH or IMS followed by real-time PCR were 10,000-fold more sensitive than direct real-time PCR when tested using boiled bacterial cell suspensions, with detection thresholds of 10(1) CFU/ml compared with 10(5) CFU/ml. DNA extraction with the Powerfood DNA extraction kit was 10-fold more sensitive than direct real-time PCR, with a detection threshold of 10(4) CFU/ml. All three assays were able to detect A. vitis in healthy-appearing grapevine cuttings taken from infected vines.

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“…This method is by far the most popular RNA extraction technique and has been used successfully to extract nucleic acid of viruses in a wide range of bulb species (Gibbs et al 2002;Sato et al 2002;Sharma et al 2005;Meenakshi et al 2006;Miglino et al 2007). Nucleic acid of better quality and quantity was generally obtained from the basal sections of the bulbs.…”

Section: Discussionmentioning

confidence: 99%

Detection of five viruses infecting dormant bulbs by TaqMan-based real-time RT-PCR

Wei

Lebas

Shiller

et al. 2011

Australasian Plant Pathol.

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Dormant bulbs may be infected by a number of viruses which are of potential economic importance for the horticultural industry. Five important viruses in bulbs were successfully detected by TaqMan-based real-time one-step RT-PCR assays: Arabis mosaic virus (ArMV), Cucumber mosaic virus (CMV), Lily symptomless virus (LSV), Tobacco rattle virus (TRV) and Tulip virus X (TVX). Nucleic acid of appropriate quality and quantity for RT-PCR was extracted from nine bulb species using the Qiagen RNeasy plant mini kit. Depending on the assay, the sensitivity of the real-time RT-PCRs varied from being the same sensitivity to 1,000 times more sensitive than the respective conventional RT-PCRs. The reliability of the TVX real-time RT-PCR was further assessed against conventional RT-PCR using leaves and bulbs of 24 Tulipa × hybrida plants. TVX was detected in twice the number of leaf samples than bulb samples for both conventional and real-time RT-PCRs.

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A semi-automated and highly sensitive streptavidin magnetic capture-hybridization RT-PCR assay: Application to genus-wide or species-specific detection of several viruses of ornamental bulb crops

Cited by 7 publications

References 28 publications

Tobraviruses—plant pathogens and tools for biotechnology

Tobraviruses—plant pathogens and tools for biotechnology

Development of a Magnetic Capture Hybridization Real-Time PCR Assay for Detection of TumorigenicAgrobacterium vitisin Grapevines

Detection of five viruses infecting dormant bulbs by TaqMan-based real-time RT-PCR

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