Detection of Cauliflower Mosaic Virus in Leaf Extracts, Protoplasts and Aphids by Enzyme-linked Immunosorbent Assay (Elisa)

1981 ◽  
Vol 100 (3) ◽  
pp. 270-278 ◽  
Author(s):  
D. H. Plessis ◽  
M. Barbara Wechmar
Keyword(s):  
Mosaic Virus ◽  
Immunosorbent Assay ◽  
Cauliflower Mosaic Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Leaf Extracts

scholarly journals Reservoir Weed Hosts for Turnip mosaic virus in Iran

Plant Disease ◽  
2005 ◽  
Vol 89 (3) ◽  
pp. 339-339 ◽  
Author(s):  
Sh. Farzadfar ◽  
K. Ohshima ◽  
R. Pourrahim ◽  
A. R. Golnaraghi ◽  
S. Sajedi ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Plant Viruses ◽  
Turnip Mosaic Virus ◽  
Cauliflower Mosaic Virus ◽  
Universal Primers ◽  
Enzyme Linked Immunosorbent Assay ◽  
Natural Occurrence ◽  
Leaf Extracts ◽  
Chenopodium Amaranticolor ◽  
Weed Hosts

During the summer of 2003, weed samples of Rapistrum rugosum and Sisymbrium loeselii showing severe mosaic, malformation, and stunting were collected from cauliflower fields in Tehran Province of Iran. Using double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) with specific polyclonal antibodies, the samples were tested for the presence of Beet western yellows virus, Cauliflower mosaic virus, Radish mosaic virus, Turnip crinkle virus, Turnip mosaic virus (TuMV) (DSMZ, Braunschweig, Germany), Cucumber mosaic virus, and Tobacco mosaic virus (Sanofi Diagnostics Pasteur, Marnes-La-Coquette, France). Leaf extracts were used for mechanical inoculation and they produced chlorotic local lesions on Chenopodium amaranticolor, necrotic lesions on leaves and shoot apex necrosis on Nicotiana glutinosa, leaf deformation, mosaic, and stunting on Petunia hybrida, and severe mosaic, distortion, and stunting on Brassica rapa. These symptoms were similar to those that were described previously for TuMV (4). ELISA results showed that the original leaf samples and inoculated indicator plants reacted positively with TuMV antibodies, but not with antibodies for any of the other viruses listed above. Also, reverse transcription-polymerase chain reaction of total RNA extracted from the collected leaf samples using the universal primers for potyviruses (3) resulted in the amplification of two fragments of the expected sizes, approximately 700 and 1,700 bp. TuMV, a member of the genus Potyvirus in the family Potyviridae, is transmitted by aphids in a nonpersistent manner (4). This virus is geographically widespread with a wide host range that can infect 318 species in 156 genera of 43 plant families including, Brassicaceae, Chenopodiaceae, Asteraceae, Cucurbitaceae, and Solanaceae (2,4). R. rugosum and S. loeselii, two annual or biennial plants in the Brassicaceae family, were common and widely distributed in the fields surveyed. The presence of TuMV-infected weed hosts in cauliflower fields may impact disease management strategies. TuMV was first observed on stock plants (Matthiola sp.) in Iran (1). To our knowledge, this is the first report of natural occurrence of TuMV on weed hosts in Iran. References: (1) M. Bahar et al. Iran. J. Plant Pathol. 21:11, 1985. (2) J. R. Edwardson and R. G. Christie. The potyvirus group. Fla. Agric. Exp. Stn. Monogr. Ser. No. 16, 1991. (3) A. Gibbs and A. Mackenzie. J. Virol. Methods 63:9, 1997. (4) J. A. Tomlinson. Turnip mosaic virus. No. 8 in: Descriptions of Plant Viruses. CMI/AAB, Surrey, England, 1970.

scholarly journals 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

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Saengsoon Charoenvilaisiri ◽  
Channarong Seepiban ◽  
Mallika Kumpoosiri ◽  
Sombat Rukpratanporn ◽  
Nuchnard Warin ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Immunosorbent Assay ◽  
Mosaic Disease ◽  
Cassava Mosaic Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sri Lankan ◽  
Stem Cuttings ◽  
Field Samples ◽  
Cassava Stem ◽  
Cassava Mosaic Virus

Abstract 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.

scholarly journals Avaliação da reação de genótipos de alface (Lactuca sativa L.) ao Lettuce mosaic virus (LMV)

Bragantia ◽  
2007 ◽  
Vol 66 (1) ◽  
pp. 61-68 ◽  
Author(s):  
Rosa Maria Chung ◽  
Joaquim Adelino de Azevedo Filho ◽  
Addolorata Colariccio
Keyword(s):  
Mosaic Virus ◽  
Immunosorbent Assay ◽  
Lactuca Sativa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Lettuce Mosaic Virus ◽  
Lactuca Sativa L

O trabalho teve como meta avaliar a reação de 18 linhagens superiores do programa de melhoramento de alface (Lactuca sativa L.) do IAC e de seis cultivares comerciais, ao Lettuce mosaic virus (LMV). Em condições de campo, na região de Atibaia (SP), foram observados sintomas de mosaico, nanismo e necrose em plantas das cultivares Rider, 'Karla H25' e Hortência. O vírus presente nos isolados foi identificado por meio de inoculação mecânica em plantas indicadoras e diferenciadoras e de testes sorológicos de Plate Trapped Antigen-Enzyme linked-immunosorbent assay (PTA-ELISA). Nas amostras avaliadas, identificou-se a espécie LMV pelo PTA-ELISA e do patotipo IV pela reação nas hospedeiras diferenciais. Para a avaliação do comportamento dos genótipos de alface, foi empregado o LMV isolado 'Karla H25'. Foram submetidos à inoculação 24 genótipos de alface empregando-se, como controle positivo, a alface 'White Boston' por sua suscetibilidade ao LMV. O delineamento experimental foi inteiramente ao acaso e analisado pelo teste do qui-quadrado. Detectaram-se genótipos com comportamento de suscetibilidade e de tolerância. Nos genótipos 3 e 4, foram observadas plantas com comportamento de tolerância ao LMV isolado 'Karla H25', enquanto nos demais genótipos, constataram-se plantas com comportamento suscetível. O plantio de cultivares tolerantes pode ser uma alternativa aos prejuízos causados pela infecção pelo LMV com conseqüente diminuição do uso de produtos químicos para o controle dos afídeos vetores.

Storage of Wheat FoliagePrior to Enzyme-Linked Immunosorbent Assay for Detection of Wheat Soil-Borne Mosaic Virus

1989 ◽  
Vol 127 (2) ◽  
pp. 116-122 ◽  
Author(s):  
C. R. Armitage ◽  
R. M. Hunger ◽  
J. L. Sherwood ◽  
D. L. Weeks
Keyword(s):  
Mosaic Virus ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay

An enzyme-linked immunosorbent assay for the detection of apple mosaic virus in yellow birch

1980 ◽  
Vol 10 (3) ◽  
pp. 278-283 ◽  
Author(s):  
T. Hardcastle ◽  
A. R. Gotlieb
Keyword(s):  
Mosaic Virus ◽  
Immunosorbent Assay ◽  
Leaf Tissue ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Method ◽  
Yellow Birch ◽  
Field Samples

An enzyme-linked immunosorbent assay (ELISA) method was developed to detect apple mosaic virus (ApMV) in Betulaalleghaniensis Britton. Tests for virus using bark and bud tissues of dormant trees were successful. ApMV was also detectable in old leaf tissue in August and September, as well as in newly emerging leaf tissue forced in a greenhouse in March. Whole crude antiserum used to coat ELISA plates in tests with bud tissues was a reliable substitute for purified immunoglobulin without loss of sensitivity or specificity. An attempt was made to use ELISA for quantifying virus concentrations in field samples of ApMV-infected birch leaves.

Effect of chemical seed treatments on symptoms caused by seed-borne barley stripe mosaic virus in Vantage barley

1986 ◽  
Vol 32 (2) ◽  
pp. 189-192 ◽  
Author(s):  
R. Vincent Miller ◽  
Thomas W. Carroll ◽  
David C. Sands
Keyword(s):  
Mosaic Virus ◽  
Immunosorbent Assay ◽  
Virus Antigen ◽  
Enzyme Linked Immunosorbent Assay ◽  
Barley Stripe Mosaic Virus ◽  
Seed Treatments ◽  
Chemical Treatments

A protocol was developed for testing chemical treatments for activity against seed-borne barley stripe mosaic virus. Vantage barley seeds infected with the MI-3 strain of the virus were allowed to imbibe dimethylsulfoxide solutions of test compounds. Of 49 compounds tested, alone or in combination with butylated hydroxyanisol, 18 reduced the number of plants expressing viral symptoms by up to 50%. Using an enzyme-linked immunosorbent assay to detect barley stripe mosaic virus, less than 2% of the asymptomatic plants arising from five chemical treatments were found to contain detectable virus antigen. In some treatments, reductions in the number of emerged plants with virus symptoms were correlated with reduced emergence.

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