Broad-spectrum detection and quantitation methods of Soil-borne cereal mosaic virus isolates

Vaïanopoulos, Céline; Legrève, Anne; Moreau, Virginie; Bragard, Claude

doi:10.1016/j.jviromet.2009.03.026

Cited by 8 publications

(4 citation statements)

References 33 publications

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“…A number of commercial tests using enzyme-linked immunosorbent assay (ELISA) and/or immunocapture polymerase chain reaction (IC-PCR) are available for the detection of viruses such as BSMV and RGMV (Ãy et al, 2008). Virus-specific reverse transcription polymerase chain reaction (RT-PCR) assays are also available for some members of these genera but not all (Chen et al, 1996;Salm et al, 1996;Webster et al, 1996;Diao et al, 1999;Clover et al, 2001;Koenig et al, 2002;Budge et al, 2008;Vaïanopoulos et al, 2009). RT-PCR assays utilising broad-range primers have been demonstrated to provide a faster tool for the detection of multiple viruses, and broad detection is also more likely to detect unreported species (Ha et al, 2008;Zheng et al, 2010).…”

Section: Q5mentioning

confidence: 99%

Novel genus-specific broad range primers for the detection of furoviruses, hordeiviruses and rymoviruses and their application in field surveys in South-east Australia

Zheng

Tang

Clover

et al. 2015

Journal of Virological Methods

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A number of viruses from the genera Furovirus, Hordeivirus and Rymovirus are known to infect and damage the four major temperate cereal crops, wheat, barley, sorghum and oats. Currently, there is no active testing in Australia for any of these viruses, which pose a significant biosecurity threat to the phytosanitary status of Australia's grains industry. To address this, broad spectrum PCR assays were developed to target virus species within the genera Furovirus, Hordeivirus and Rymovirus. Five sets of novel genus-specific primers were designed and tested in reverse-transcription polymerase chain reaction assays against a range of virus isolates in plant virus diagnostic laboratories in both Australia and New Zealand. Three of these assays were then chosen to screen samples in a three-year survey of cereal crops in western Victoria, Australia. Of the 8900 cereal plants screened in the survey, all were tested free of furoviruses, hordeiviruses and rymoviruses. To date, there were no published genus-specific primers available for the detection of furoviruses, hordeiviruses and rymoviruses. This study shows for the first time a broad-spectrum molecular test being used in a survey for exotic grain viruses in Australia. Results from this survey provide important evidence of the use of this method to demonstrate the absence of these viruses in Victoria, Australia. The primer pairs reported here are expected to detect a wide range of virus species within the three genera.

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Section: Q5mentioning

confidence: 99%

Novel genus-specific broad range primers for the detection of furoviruses, hordeiviruses and rymoviruses and their application in field surveys in South-east Australia

Zheng

Tang

Clover

et al. 2015

Journal of Virological Methods

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“…Further, it was effective in detecting the isolates of SBCMV from Belgium, France, Germany, Italy and United Kingdom. The real-time RT-PCR protocol showed promise for reliable comparison of soil inoculum potential as well as for determining levels of cultivar resistance to SBCMV (Vaïanopoulos et al 2009). …”

Section: ) (Appendix 3)mentioning

confidence: 99%

Detection of Viral and Viroid Plant Pathogens in the Environment

Narayanasamy¹

2010

Microbial Plant Pathogens-Detection and Disease Diagnosis:

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Plant viruses and viroids with obligate parasitism as the mode of existence, have to be in association with living cells of plant hosts always. However, some highly stable viruses have been detected in a free state in soil, water or air. Other viruses depend on the vectors such as insects, mites, nematodes or fungi present in the environment for their spread from infected plants to healthy plants which may be available in the same field or far away. The role of alternative/additional host plant species functioning as sources of virus infection has been demonstrated by applying various diagnostic techniques. The presence of viruses has been detected by employing biological, immunological and nucleic acid-based techniques. The effectiveness and reliability of detection varies significantly. However, molecular techniques have been shown to be more sensitive, specific and fast in providing the results, although they are unable to indicate the level of pathogenic potential of the viruses detected in the environmental samples. Viroids lacking natural vectors and stability outside living cells have been detected in several additional/alternative plant species which may function as reservoir of viroid infection.

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“…This high copy number makes the 3 -UTR a logical target for highly efficient broad-spectrum detection. This strategy had already been used for detecting another virus also transmitted by Polymyxa, Soil-borne cereal mosaic virus (Vaianopoulos et al, 2009). RT-PCR using J8-J9 primer pairs allowed all the currently known IPCV and PCV isolates to be detected.…”

Section: Broad-spectrum Rt-pcr For Detection and Quantitationmentioning

confidence: 99%

Targeting highly conserved 3′-untranslated region of pecluviruses for sensitive broad-spectrum detection and quantitation by RT-PCR and assessment of phylogenetic relationships

Dieryck

Delfosse

Reddy

et al. 2010

Journal of Virological Methods

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The 3'-end region of many virus isolates has been shown to possess conserved sequences in addition to the presence of numerous genomic and subgenomic RNAs. Utilizing these sequences, a broad-spectrum reverse transcription-polymerase chain reaction protocol has been developed to detect all the known Indian peanut clump virus and Peanut clump virus isolates, that cause peanut clump diseases in West Africa and India. The primers were targeted at the highly conserved 3'-untranslated regions of the PCV RNA-1 and RNA-2. The conservation was confirmed by sequencing these untranslated regions of RNA-1 for six isolates and RNA-2 for one isolate. The conserved structure of the RNA-1 and RNA-2 was observed and the importance of this region for the virus survival was confirmed. The primers were also designed for virus quantitation using a Taqman(®)-based real-time RT-PCR. The use of RT-PCR and real-time quantitative RT-PCR improved the sensitivity of PCV detection compared to ELISA. RT-PCR also led to the detection of IPCV and PCV on two new natural hosts: Oldenlandia aspera and Vigna subterranea. Real-time RT-PCR is considered to be an ideal tool for identifying resistant sources to both IPCV and PCV.

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Broad-spectrum detection and quantitation methods of Soil-borne cereal mosaic virus isolates

Cited by 8 publications

References 33 publications

Novel genus-specific broad range primers for the detection of furoviruses, hordeiviruses and rymoviruses and their application in field surveys in South-east Australia

Novel genus-specific broad range primers for the detection of furoviruses, hordeiviruses and rymoviruses and their application in field surveys in South-east Australia

Detection of Viral and Viroid Plant Pathogens in the Environment

Targeting highly conserved 3′-untranslated region of pecluviruses for sensitive broad-spectrum detection and quantitation by RT-PCR and assessment of phylogenetic relationships

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