Development of Routine Rt-PCR Elosa Tests for Fruit Tree Certification

Kummert, J.; Vendrame, Marina; Lepoivre, Philippe; Steyer, Stéphane

doi:10.17660/actahortic.2001.550.3

Cited by 11 publications

(3 citation statements)

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“…Specific primers for PNRSV were identified by computer analysis using the PILEUP, FASTA, and PRIME programs (Wisconsin Package version 10.1; Genetic Computer Group, Madison, WI). These programs were applied to sequence data corresponding to the virus coat protein (CP)-encoding sequence of PNRSV-RNA 4, from 24 isolates available in the GenBank database of National Center for Biotechnology Information and at the European Molecular Biology Laboratory databases (11). Among the potential primer pairs identified, the primers PNRSV 10F (TTC TTG AAG GAC CAA CCG AGA GG) and PNRSV 10R (GCT AAC GCA GGT AAG ATT TCC AAG C), amplifying a fragment of 348 bp, were selected.…”

Section: Methodsmentioning

confidence: 99%

Development of Real-Time RT-PCR Assay for Detection of Prunus necrotic ringspot virus in Fruit Trees

et al. 2003

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A real-time fluorescent reverse-transcriptase polymerase chain reaction (RT-PCR) assay using a short fluorogenic 3′ minor groove binder (MGB) DNA hydrolysis probe was developed for the detection of Prunus necrotic ringspot virus (PNRSV) in stone fruit trees. The covalent attachment of the minor groove binder moiety at the 3′ end of the probe increased the probe target duplex stability and raised the melting temperature to a range suitable for real-time analysis. The real-time RT-PCR assay correlated well with conventional RT-PCR results for the detection of PNRSV. This assay reliably detects PNRSV in bark tissues of dormant cherry and plum trees. Furthermore, it is well adapted for the routine detection of PNRSV because it eliminates one risk of contamination by performing the whole test in a single closed tube. This system may replace the commonly used diagnostic techniques (e.g., woody indicators and immunological tests) to detect this virus.

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Section: Methodsmentioning

confidence: 99%

Development of Real-Time RT-PCR Assay for Detection of Prunus necrotic ringspot virus in Fruit Trees

et al. 2003

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“…This is probably because the development of a multiplex RT-PCR system can be laborious and time consuming, but is a rapid, reliable, and cost-effective method (Wei et al, 2008). RT-PCR has been successfully used for simultaneous detection of several stone fruit viruses, even with low virus titre and in the presence of inhibitors (Bariana et al, 1994;Kummert et al, 2001). Therefore, focus in the present study was placed on the results obtained with RT-PCR.…”

Section: Discussionmentioning

confidence: 99%

Occurrence of cherry viruses in South Tyrol (Italy) by comparing growth periods in two consecutive years

Deltedesco¹,

NIEDRIST²,

Oettl³

2022

Phytopathol. Mediterr.

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Sweet cherries (Prunus avium L.) are important as fruit crops, and can be affected by numerous viruses. An investigation on the occurrence of the three most common viruses of sweet cherry was carried out in commercially managed orchards in South Tyrol (Italy). The incidence of apple chlorotic leaf spot virus (ACLSV), Prunus necrotic ringspot virus (PNRSV) and prune dwarf virus (PDV) was investigated using enzyme-linked immunosorbent assays (ELISA) and the reverse transcriptase-polymerase chain reaction technique (RT-PCR) in spring 2018 and 2020, and during the summer and autumn of 2020. All three viruses were detected in the surveyed orchards. Comparative analyses showed that detection was more effective with RT-PCR than with ELISA, especially for detecting PNRSV and PDV. Mixed infections were detected in all the surveyed orchards. The results also showed clear differences between and during host growth periods, likely due to a variable virus concentration in the host trees.

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“…Therefore, molecular hybridization based techniques represent additional alternative for biological and serological assays. Polymerase chain reaction (PCR) has contributed to increasing the detection sensitivity for low virus titre or for the presence of inhibitors (Bariana et al, 1995;Rowhani et al, 1995;Kummert et al, 2001). Reverse transcription-PCR (RT-PCR) has the potential to be an extremely sensitive method in some woody plants even during seasons of low titre of virus (Kinard et al, 1996;Spiegel et al, 1996;Sanchez-Navarro et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

Simultaneous detection of Apple mosaic virus in cultivated hazelnuts by one-tube RT-PCR

Akbaş¹,

Deği̇rmenci̇²

2010

Afr. J. Biotechnol.

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The most economically damaging ilarvirus affecting hazelnut on a worldwide scale is the related apple mosaic virus (ApMV). Attempts were made to isolate the virus RNA from hazelnut tissues using different extraction methods. The most suitable extraction method that could detect the virus occurring naturally in hazelnut by reverse-transcription polymerase chain reaction (RT-PCR) methodology was selected. RT-PCR was applied successfully using flower, husk and leaf tissues. The most suitable extraction method and hazelnut tissues determined were sensitive, simple, rapid and reliable for simultaneous detection of ApMV in hazelnut tissues. To our knowledge, this is the first report of the simultaneous detection of the virus by RT-PCR, an alternative detection of ApMV in hazelnut hosts.

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Development of Routine Rt-PCR Elosa Tests for Fruit Tree Certification

Cited by 11 publications

References 0 publications

Development of Real-Time RT-PCR Assay for Detection of Prunus necrotic ringspot virus in Fruit Trees

Development of Real-Time RT-PCR Assay for Detection of Prunus necrotic ringspot virus in Fruit Trees

Occurrence of cherry viruses in South Tyrol (Italy) by comparing growth periods in two consecutive years

Simultaneous detection of Apple mosaic virus in cultivated hazelnuts by one-tube RT-PCR

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