Detection of Potato Leafroll and Strawberry Mild Yellow-Edge Luteoviruses by Reverse Transcription-Polymerase Chain Reaction Amplification

Hadidi, A.; Montasser, Magdy S.; Lévy, L.; Goth, R. W.; Converse, R. H.; Madkour, M. A.; Skrzeckowski, L. J.

doi:10.1094/pd-77-0595

Cited by 42 publications

(17 citation statements)

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“…Conventional polymerase chain reaction (PCR) or reverse transcription-PCR (RT-PCR) which have high sensitivity and specificity over ELISA-based methods and have been used to detect DNA viruses or RNA viruses, respectively (Bostan and Peker, 2009;Stark et al, 2008;Zaghloul, 2011). RT-PCR and immunocapture reverse transcription polymerase chain reaction (IC-RT-PCR) have been employed for detection of PLRV in plant or in aphid vectors (Ahouee et al, 2010;Hadidi et al, 1993;Peiman and Xie, 2006;Singh et al, 1995;1997). Notomi et al (2000) developed loop-mediated isothermal amplification (LAMP) to amplify DNA with high specificity.…”

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confidence: 99%

“…RT-PCR (Hadidi et al, 1993;Peiman and Xie, 2006;Singh et al, 1995;1997) and IC-RT-PCR (Ahouee et al, 2010) have been developed for the rapid and accurate detection of PLRV in plants or/and aphids. Although these conventional PCR-based methods increased rapidness and accuracy for PLRV detection, these methods require expensive and sophisticated instruments for the PLRV amplification and detection of the PCR products.…”

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confidence: 99%

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Simple and Rapid Detection of Potato leafroll virus by Reverse Transcription Loop-mediated Isothermal Amplification

Ju¹

2011

The Plant Pathology Journal

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A new reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for the Potato leafroll virus (PLRV) was developed and compared with conventional reverse transcription polymerase chain reaction (RT-PCR) to address its advantages over RT-PCR. RT-LAMP primers were designed from the open reading frame 3 (ORF3) sequence of PLRV. The RT-LAMP reactions were conducted without or with a set of loop primers. By real-time monitoring using Turbimeter, the RT-LAMP (with loop primers) detects PLRV in less than 30 min, compared to 120 min of RT-PCR. By adding fluorescent reagent during the reaction, final products of the RT-LAMP were fluorescently visualized under UV light or could be differentiated by naked-eye inspection under normal light. The RT-LAMP was extremely sensitive, about 2000-fold more sensitive than RT-PCR. This study presents great potential of the RT-LAMP for diagnosis and PLRV epidemiology because RT-LAMP method is speedy, sensitive, inexpensive, and convenient.

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confidence: 99%

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Simple and Rapid Detection of Potato leafroll virus by Reverse Transcription Loop-mediated Isothermal Amplification

Ju¹

2011

The Plant Pathology Journal

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“…The oligo-nucleotide primers were synthesized in Thermo Hybaid GmbH, Germany. Viral cDNA was synthesized and amplified as procedure according to Hadidi et al (1993). One µg of RNA, 3 ml of the primers, 6 µl of 5X first strand cDNA buffer (250 mMTris-HCl, pH 8.3; 500 mMKCl; 15 mM MgCl 2 ), 3 µl of 0.1 Mdithiothreitol (DTT), were added to a final volume of 30 µl by deionized water.…”

Section: Molecular Characters Reverse Transcription-polymerase Chain mentioning

confidence: 99%

Physiological and Infectious Characters of Potato virus Y-Egyptian isolate

Abdel-Shafi

Ghaly

El-Dougdoug

et al. 2017

Egypt. J. Microbiol.

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POTATO VIRUS Y (PVY) is one of the most destructive aphid transmitted pathogen to potato plants worldwide. In Egypt, PVY infection causing about 80% reduction the global yield of potato. The objective of this study was to characterize Potato virus Y (PVY-EG) infecting potato plants, based on biological, serological and molecular properties. Naturally infected potato plants by PVY gave positive reaction with PVY-polyclonal antibodies using DAS-ELISA for virus identification. PVY has a higher stability at 45 h (Longevity), 66°C at 10 min (TIP) at 10 -6 (DEP), revealing presence of cytoplasmic and crystalline inclusion bodies of the epidermal strips from infected Datura metel (diagnostic host) leaves at 12 days post PVY inoculation. The PVY has UV-spectra at λ 235 and at λ 257 nm. PVY yield was 1.25/100 g leaf tissues and 260/280 more than 1 (it was 1.6). The viral particles were rod flexible (helical symmetry) of 11 x 570 nm, with obvious immunogenicity that represented by 1:1024 titer of antibodies. The cDNA fragment of CP gene was 610 bp. Sequence analysis revealed that PVY isolate showed 93-99% similarity with other worldwide PVY isolates.

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“…For example, various equipment exists to automate the different coating and washing stages, and reading and recording of test results. It is currently the method of choice for testing microplants and growing crop inspection leaf samples for the presence of potato virus (Browning, 1995;Hadidi et al, 1993;Rose & Hubbard, 1986;Schroeder & Weidemann, 1990).…”

Section: Serological Diagnosticsmentioning

confidence: 99%

Plant pathogen diagnostics: present status and future developments

O'Donnell

1999

Potato Res

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Detection of Potato Leafroll and Strawberry Mild Yellow-Edge Luteoviruses by Reverse Transcription-Polymerase Chain Reaction Amplification

Cited by 42 publications

References 0 publications

Simple and Rapid Detection of Potato leafroll virus by Reverse Transcription Loop-mediated Isothermal Amplification

Simple and Rapid Detection of Potato leafroll virus by Reverse Transcription Loop-mediated Isothermal Amplification

Physiological and Infectious Characters of Potato virus Y-Egyptian isolate

Plant pathogen diagnostics: present status and future developments

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