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.

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.

scholarly journals First Report of Soilborne Wheat Mosaic Virus in the United Kingdom

Plant Disease ◽  
1999 ◽  
Vol 83 (9) ◽  
pp. 880-880 ◽  
Author(s):  
G. R. G. Clover ◽  
D. M. Wright ◽  
C. M. Henry
Keyword(s):  
United Kingdom ◽  
Mosaic Virus ◽  
Barley Yellow Dwarf Virus ◽  
Protein A ◽  
The United States ◽  
Enzyme Linked Immunosorbent Assay ◽  
Barley Yellow Dwarf ◽  
The United Kingdom ◽  
Field Samples ◽  
Yellow Dwarf Virus

In April 1999, severe soilborne wheat mosaic virus (SBWMV) symptoms were observed in five fields of winter wheat (Triticum aestivum, cvs. Consort, Equinox, and Savannah) on one farm in Wiltshire, UK. Affected plants were markedly stunted and had a pale mosaic on their leaf sheaths that developed into bright yellow, parallel streaks on the leaves as they unfolded. Symptomatic plants were found in discrete, elliptical patches ranging in size from a few square meters to nearly a hectare. During May and June, symptoms became less marked as temperatures increased and were restricted to lower leaves. SBWMV was positively identified in all five fields (60 to 170 plants per field) by double (W. Huth, BBA-Braunschweig, Germany; Sanofi Phyto-Diagnostics, Paris) and triple (T. Wilson, SCRI, Dundee, UK) antibody sandwich enzyme-linked immunosorbent assay and by reversetranscription polymerase chain reaction (2). Identification was confirmed by immunoelectron microscopy, including protein-A gold labeling, which revealed bipartite, rod-shaped particles typical of SBWMV. Neither wheat spindle streak mosaic virus nor barley yellow dwarf virus was detected in the field samples, nor was SBWMV detected in any other field subsequently sampled, despite a survey of the surrounding area. Wheat is the most important economic crop in the United Kingdom (≈1.9 million hectares are grown annually, yielding ≈16 million tonnes), but its position is threatened by the economic impact of SBWMV, which has decreased yields by up to 50% in the United States (1). References: (1) T. A. Kucharek and J. H. Walker. Plant Dis. Rep. 58:763, 1974. (2) R. E. Pennington et al. Plant Dis. 77:1202, 1993.

scholarly journals Development of an antigen-capture enzyme-linked immunosorbent assay for Clostridium perfringens beta2-toxin in porcine feces and the neonatal piglet intestine

2012 ◽  
Vol 24 (5) ◽  
pp. 895-902 ◽  
Author(s):  
Jasmina Kircanski ◽  
Douglas Hodgins ◽  
Glenn Soltes ◽  
Yanlong Pei ◽  
Valeria R. Parreira ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Protease Activity ◽  
Limit Of Detection ◽  
Intestinal Content ◽  
Enzyme Linked Immunosorbent Assay ◽  
Neonatal Piglet ◽  
Antigen Capture ◽  
Field Samples ◽  
Intestinal Contents ◽  
Beta2 Toxin

An enzyme-linked immunosorbent assay (ELISA) was developed for detection and quantitation of beta2-toxin in neonatal piglet intestinal contents. Polystyrene plates were coated with polyclonal capture antibodies prepared against consensus recombinant beta2-toxin. The ELISA was developed using consensus recombinant beta2-toxin, atypical recombinant beta2-toxin, purified consensus native beta2-toxin, and field samples of neonatal porcine intestinal contents. Captured antigen was detected using a horseradish peroxidase–labeled monoclonal antibody against consensus recombinant beta2-toxin. The limit of detection of the ELISA for consensus beta2-toxin was between 2.0 and 3.5 ng/ml. The ELISA detected atypical recombinant beta2-toxin only weakly. Optical density was protein concentration dependent. The test confirmed differences between consensus and atypical recombinant beta2-toxin, but similar results obtained when testing pure consensus recombinant beta2-toxin and native beta2-toxin. Results obtained from intestinal content samples, particularly from the small intestine, were highly inconsistent and suggested variable protease activity. Addition of protease inhibitors partially prevented degradation of the toxin; however, sample processing at low temperature, at a lower pH (citrate buffer with 5% of bovine serum albumin, pH 6.1), and “cold incubation” of applied antigens abolished protease activity. The recombinant toxin was preserved in spiked intestinal samples by freezing at −70°C, suggesting that necropsy samples can be stored frozen for periodic testing. With appropriate sample preparation, antigen-capture ELISA can detect beta2-toxin in the intestinal content and feces of neonatal piglets.

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