Enhanced Detection of Prune Dwarf Virus in Peach Leaves by Immunocapture‐Reverse Transcription‐Polymerase Chain Reaction with Nested Polymerase Chain Reaction (IC‐RT‐PCR Nested PCR)

Helguera, Pablo; Docampo, Delia M.; Nomé, S. F.; Ducasse, Daniel Adrián

doi:10.1046/j.1439-0434.2002.00696.x

Cited by 10 publications

(2 citation statements)

References 7 publications

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“…Such is the case for Prunus necrotic ringspot virus, Prune dwarf virus, Potato virus Y and Beet yellows virus; an increase in detection was achieved with IC-RT-PCR or adding nested PCR after the IC-RT-PCR (Stevens et al, 1997;Sanchez-Navarro et al, 1998;Moury et al, 2000;Varveri, 2000;Helguera et al, 2001Helguera et al, , 2002. Here we used this method at a crucial stage in the production of virus-free garlic plants, when the plants are still in the in vitro culture phase and the concentration of virus is generally very low.…”

Section: Discussionmentioning

confidence: 99%

Improved PCR detection of potyviruses in Allium species

Lunello¹,

Ducasse

Conci

2005

Eur J Plant Pathol

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Protocols for producing virus-free Allium plants require an indexing system that is more sensitive than DAS-ELISA and can detect low virus concentrations in infected plants. In the present work, degenerate primers were designed and a one-step IC-RT-PCR protocol was developed to differentiate between Leek yellow stripe virus (LYSV) and Onion yellow dwarf virus (OYDV) in single and mixed infections in several Allium spp. A 566-bp band was observed for LYSV, a 489-bp band for OYDV in single infections, and two bands of the same sizes in mixed infections in different species of Alliaceae. A 508-bp band of Shallot yellow stripe virus and a 594-bp band of Turnip mosaic virus were also amplified with the same primers. RTnested-PCR was also conducted directly in microtitre plate wells after negative or questionable reactions were produced in an ELISA experiment. The detection limit of the DAS-ELISA for LYSV was 16.5-27.3 ng ml )1 . The RT-nested-PCR done after DAS-ELISA was 10 2 times more sensitive than the DAS-ELISA alone. In parallel, an IC-RT-nested-PCR in microcentrifuge tubes was 10 4 times more sensitive than the DAS-ELISA. The DAS-ELISA-RT-nested-PCR enables the initial screening of samples by DAS-ELISA to eliminate a high percentage of virus-positive plants, considerably reducing the number of plants to analyze further by RT-PCR.

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Section: Discussionmentioning

confidence: 99%

Improved PCR detection of potyviruses in Allium species

Lunello¹,

Ducasse

Conci

2005

Eur J Plant Pathol

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“…Problems mentioned above can be solved by Nested PCR (Olmos et al, 1999). Several viruses, including PNRSV, PDV, PPV and CTV, were detected by this technique (Adkar- Purushothama et al, 2011;Helguera et al, 2001Helguera et al, , 2002Olmos et al, 1999). This nested PCR was combine with Immunocapture-RT PCR to increase sensitivity and to simplify preparation of sample (Helguera et al, 2001(Helguera et al, , 2002.…”

Section: Polymerase Chain Reaction (Pcr) Pcr and Reverse Transcriptimentioning

confidence: 99%

A Review of Detection Methods for the Plant Viruses

Jeong¹,

Ju²,

Noh³

2014

Research in Plant Disease

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The early and accurate detection of plant viruses is an essential component to control those. Because the globalization of trade by free trade agreement (FTA) and the rapid climate change promote the country-tocountry transfer of viruses and their hosts and vectors, diagnosis of viral diseases is getting more important. Because symptoms of viral diseases are not distinct with great variety and are confused with those of abiotic stresses, symptomatic diagnosis may not be appropriate. From the last three decades, enzyme-linked immunosorbent assays (ELISAs), developed based on serological principle, have been widely used. However, ELISAs to detect plant viruses decrease due to some limitations such as availability of antibody for target virus, cost to produce antibody, requirement of large volume of sample, and time to complete ELISAs. Many advanced techniques allow overcoming demerits of ELISAs. Since the polymerase chain reaction (PCR) developed as a technique to amplify target DNA, PCR evolved to many variants with greater sensitivity than ELISAs. Many systems of plant virus detection are reviewed here, which includes immunological-based detection system, PCR techniques, and hybridization-based methods such as microarray. Some of techniques have been used in practical, while some are still under developing to get the level of confidence for actual use.

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