Detection of geminiviruses from tropical countries by a double monoclonal antibody ELISA using antibodies to African cassava mosaic virus

Givord, Louise; Fargette, Denis; Kounounguissa, B.; Thouvenel, J.C.; Walter, Bernard; Regenmortel, Mhv Van

doi:10.1051/agro:19940506

Cited by 12 publications

(6 citation statements)

References 26 publications

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“…The efficiency of plant inoculation using the particle inflow gun was affected by several factors including plant age, gas pressure, the distance between the particle inflow gun and the plant as well as the source of the nucleic acid used for inoculation [ 6 , 49 , 50 , 51 ]. In this study the direct bombardment of plants with RCA products has shown to be a convenient and quicker tool for biolistic inoculation, in agreement with previous studies that successfully used RCA products for biolistic inoculation of geminiviruses [ 27 , 28 ].…”

Section: Resultssupporting

confidence: 89%

“…All samples produced an RCA product, and 73 showed an RFLP digestion pattern similar to the positive control and in silico digestion of WmCSV [ 27 ] ( Figure 2 ), while the rest of the samples (79) gave a different pattern similar to SLCV digestion pattern. ELISA test using virus-specific antibodies [ 28 , 29 , 30 ] and polymerase chain reaction (PCR) using either specific or degenerated primers [ 31 , 32 , 33 ] were the ordinary methods used for detection of geminiviral infection in plant samples. However, RCA combined with RFLP improved the diagnosis of geminiviral infection and was shown to be a suitable and more sensitive tool (10 ng of DNA sample would be enough using RCA) for geminiviral infection screening [ 27 , 34 , 35 , 36 , 37 ] due to the several advantages over other methods.…”

Section: Resultsmentioning

confidence: 99%

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Molecular Characterization of Watermelon Chlorotic Stunt Virus (WmCSV) from Palestine

Ali‐Shtayeh¹,

Jamous²,

Mallah³

et al. 2014

Viruses

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The incidence of watermelon chlorotic stunt disease and molecular characterization of the Palestinian isolate of Watermelon chlorotic stunt virus (WmCSV-[PAL]) are described in this study. Symptomatic leaf samples obtained from watermelon Citrullus lanatus (Thunb.), and cucumber (Cucumis sativus L.) plants were tested for WmCSV-[PAL] infection by polymerase chain reaction (PCR) and Rolling Circle Amplification (RCA). Disease incidence ranged between 25%–98% in watermelon fields in the studied area, 77% of leaf samples collected from Jenin were found to be mixed infected with WmCSV-[PAL] and SLCV. The full-length DNA-A and DNA-B genomes of WmCSV-[PAL] were amplified and sequenced, and the sequences were deposited in the GenBank. Sequence analysis of virus genomes showed that DNA-A and DNA-B had 97.6%–99.42% and 93.16%–98.26% nucleotide identity with other virus isolates in the region, respectively. Sequence analysis also revealed that the Palestinian isolate of WmCSV shared the highest nucleotide identity with an isolate from Israel suggesting that the virus was introduced to Palestine from Israel.

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Section: Resultssupporting

confidence: 89%

Section: Resultsmentioning

confidence: 99%

Molecular Characterization of Watermelon Chlorotic Stunt Virus (WmCSV) from Palestine

Ali‐Shtayeh¹,

Jamous²,

Mallah³

et al. 2014

Viruses

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“…For example the reaction of EACMV-UG was similar to ACMV (Zhou et al, 1997), as were SACMV and EACMV in ELISA (Berrie et al, 1998). Givord et al (1994) also produced two MAbs (7 and 11) against ACMV isolates from Nigeria, although these could not differentiate between ACMV and EACMV. However, the two MAbs were able to detect Begomoviruses of other crops within Africa such as Tomato yellow leaf curl virus (TomYLV) and Okra leaf curl virus (OLCV).…”

Section: Serological Characteristicsmentioning

confidence: 99%

Status of Cassava Mosaic Virus Diseases and CassavaBegomovirusesin Sub-Saharan Africa

Atiri

Ogbe

Dixon

et al. 2004

Journal of Sustainable Agriculture

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The current status of cassava Begomoviruses, the most serious constraint to the production of cassava, a major staple food crop in G. I. Atiri is Professor and Head, Please note that this electronic prepublication galley may contain typographical errors and may be missing artwork, such as charts, photographs, etc. Pagination in this version will differ from the published version.sub-Saharan Africa, is reviewed in relation to their distribution, effects, etiology, and epidemiology. It is concluded that control of the diseases would continue to depend on integrated management involving cultural practices and use of resistant cultivars. Current trends in diagnosis and control, including the production of transgenic plants, selection for resistance in cassava via molecular markers, and the determination of resistant profiles of cassava genotypes to a range of virus variants, are also discussed.

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“…Several molecular and serology-based methods have been employed for the detection of CMBs such as PCR [ 20 ], multiplex PCR [ 21 ], real-time PCR [ 22 ], immunosorbent electromicroscopy [ 23 ], double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) [ 24 ] and triple antibody sandwich-ELISA (TAS-ELISA) [ 25 ]. For SLCMV, methods such as PCR [ 26 – 28 ], multiplex PCR [ 29 , 30 ], rolling circle amplification (RCA) [ 12 ] and plate-trapped antigen-ELISA (PTA-ELISA) [ 15 ] have been described.…”

Section: Introductionmentioning

confidence: 99%

Development of a triple antibody sandwich enzyme-linked immunosorbent assay for cassava mosaic disease detection using a monoclonal antibody to Sri Lankan cassava mosaic virus

Charoenvilaisiri

Seepiban

Kumpoosiri

et al. 2021

Virol J

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Background Cassava mosaic disease (CMD) is one of the most devastating viral diseases for cassava production in Africa and Asia. Accurate yet affordable diagnostics are one of the fundamental tools supporting successful CMD management, especially in developing countries. This study aimed to develop an antibody-based immunoassay for the detection of Sri Lankan cassava mosaic virus (SLCMV), the only cassava mosaic begomovirus currently causing CMD outbreaks in Southeast Asia (SEA). Methods Monoclonal antibodies (MAbs) against the recombinant coat protein of SLCMV were generated using hybridoma technology. MAbs were characterized and used to develop a triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) for SLCMV detection in cassava leaves and stems. Assay specificity, sensitivity and efficiency for SLCMV detection was investigated and compared to those of a commercial ELISA test kit and PCR, the gold standard. Results A TAS-ELISA for SLCMV detection was successfully developed using the newly established MAb 29B3 and an in-house polyclonal antibody (PAb) against begomoviruses, PAb PK. The assay was able to detect SLCMV in leaves, green bark from cassava stem tips, and young leaf sprouts from stem cuttings of SLCMV-infected cassava plants without cross-reactivity to those derived from healthy cassava controls. Sensitivity comparison using serial dilutions of SLCMV-infected cassava sap extracts revealed that the assay was 256-fold more sensitive than a commercial TAS-ELISA kit and 64-fold less sensitive than PCR using previously published SLCMV-specific primers. In terms of DNA content, our assay demonstrated a limit of detection of 2.21 to 4.08 × 106 virus copies as determined by quantitative real-time PCR (qPCR). When applied to field samples (n = 490), the TAS-ELISA showed high accuracy (99.6%), specificity (100%), and sensitivity (98.2%) relative to the results obtained by the reference PCR. SLCMV infecting chaya (Cnidoscolus aconitifolius) and coral plant (Jatropha multifida) was also reported for the first time in SEA. Conclusions Our findings suggest that the TAS-ELISA for SLCMV detection developed in this study can serve as an attractive tool for efficient, inexpensive and high-throughput detection of SLCMV and can be applied to CMD screening of cassava stem cuttings, large-scale surveillance, and screening for resistance.

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Detection of geminiviruses from tropical countries by a double monoclonal antibody ELISA using antibodies to African cassava mosaic virus

Cited by 12 publications

References 26 publications

Molecular Characterization of Watermelon Chlorotic Stunt Virus (WmCSV) from Palestine

Molecular Characterization of Watermelon Chlorotic Stunt Virus (WmCSV) from Palestine

Status of Cassava Mosaic Virus Diseases and CassavaBegomovirusesin Sub-Saharan Africa

Development of a triple antibody sandwich enzyme-linked immunosorbent assay for cassava mosaic disease detection using a monoclonal antibody to Sri Lankan cassava mosaic virus

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