Detection of potato leafroll virus in single aphids by the reverse transcription polymerase chain reaction and its potential epidemiological application

Singh, R. P.; Kurz, Jolanta; Boiteau, Gilles; Bernard, G.

doi:10.1016/0166-0934(95)00056-z

Cited by 64 publications

(42 citation statements)

References 10 publications

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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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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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“…More sensitive techniques thus represent a possible alternative. In this respect, RT-PCR tests combining a cDNA synthesis and a PCR amplification have been described for the detection of potato viruses like PVY O , PVY NTN (Weidemann and Maiss, 1996), PLRV (Schoen et al, 1996;Singh et al, 1995) or PVA (Cerovska et al, 1998;.…”

Section: Introductionmentioning

confidence: 99%

RT-PCR-ELOSA tests on pooled sample units for the detection of virus Y in potato tubers

Chandelier

Dubois

Baelen

et al. 2001

Journal of Virological Methods

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A highly sensitive RT-PCR protocol able to detect potato virus Y (PVY) in pooled sample units (tubers) was developed. PVY-specific primers selected in the coat protein gene were found to amplify a 359 bp fragment from diluted crude extract of infected tubers. For the detection of the amplification products, a colorimetric detection procedure in microtiter plates was established. The amplicons are hybridized between a covalently linked capture probe and a specific biotinylated detection probe ELOSA tests. This detection method detects at least 50 pg of virus per reaction for the four cultivars tested. The RT-PCR-ELOSA assay was adapted to pooled units in order to increase the sample size while reducing the number of tests.

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“…A total of 9 pairs of specific primers including ones for Potato leafroll virus (PLRV) (Keese et al 1990) and Tomato spotted wilt virus (TSWV) (Mumford et al 1996) were used for detection of eight different viruses, respectively. RT-PCR was carried out with a PTC-200 thermocycler (MJ Research, Inc. USA) with reaction conditions described by Singh et al (1995). Five microlitres of amplified products were analysed by gel electrophoresis on a 1.5% agarose gel which was pre-stained with ethidium bromide and photographed under UV light at 254 nm (Gel Doc EQ, Bio-Rad).…”

mentioning

confidence: 99%

“…RNA extraction was performed mainly according to the procedure described by Singh et al (1995). Primers for the detection of several viruses were synthesised by SSBETS Co., Ltd, China.…”

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Fritillaria thumbergii—a new host ofPotato leafroll virusin China

Qin

Chen

2006

Austral. Plant Disease Notes

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Detection of potato leafroll virus in single aphids by the reverse transcription polymerase chain reaction and its potential epidemiological application

Cited by 64 publications

References 10 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

RT-PCR-ELOSA tests on pooled sample units for the detection of virus Y in potato tubers

Fritillaria thumbergii—a new host ofPotato leafroll virusin China

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