scholarly journals Fully “Recombinant Enzyme-Linked Immunosorbent Assays” Using Genetically Engineered Single-Chain Antibody Fusion Proteins for Detection of Citrus tristeza virus

2000 ◽  
Vol 90 (12) ◽  
pp. 1337-1344 ◽  
Author(s):  
Estela Terrada ◽  
Randolf J. Kerschbaumer ◽  
Giuseppe Giunta ◽  
Patrizia Galeffi ◽  
Gottfried Himmler ◽  
...  
Keyword(s):  
Fusion Proteins ◽  
Citrus Tristeza Virus ◽  
Sandwich Elisa ◽  
Viral Disease ◽  
Genetically Engineered ◽  
Enzyme Linked Immunosorbent Assay ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Tristeza Virus ◽  
Citrus Tristeza

Recombinant single-chain variable fragment antibodies (scFv) that bind specifically to Citrus tristeza virus (CTV), which cause the most detrimental viral disease in the citrus industry worldwide, were obtained from the hybridoma cell lines 3DF1 and 3CA5. These scFv were genetically fused with dimerization domains as well as with alkaline phosphatase, respectively, and diagnostic reagents were produced by expressing these fusion proteins in bacterial cultures. The engineered antibodies were successfully used for CTV diagnosis in plants by tissue print enzyme-linked immunosorbent assay (ELISA) and double antibody sandwich-ELISA. The fully recombinant ELISAs were as specific and sensitive as conventional ELISAs performed with the parental monoclonal antibodies, showing the usefulness of recombinant antibodies for routine detection of a virus in woody plants for the first time.

Transgenic expression in citrus of single-chain antibody fragments specific to Citrus tristeza virus confers virus resistance

2010 ◽  
Vol 19 (6) ◽  
pp. 1001-1015 ◽  
Author(s):  
Magdalena Cervera ◽  
Olga Esteban ◽  
Maite Gil ◽  
M. Teresa Gorris ◽  
M. Carmen Martínez ◽  
...  
Keyword(s):  
Virus Resistance ◽  
Citrus Tristeza Virus ◽  
Antibody Fragments ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Transgenic Expression ◽  
Tristeza Virus ◽  
Citrus Tristeza

scholarly journals Breakdown of Cross-Protection of Grapefruit from Decline-Inducing Isolates of Citrus tristeza virus Following Introduction of the Brown Citrus Aphid

Plant Disease ◽  
2003 ◽  
Vol 87 (9) ◽  
pp. 1116-1118 ◽  
Author(s):  
C. A. Powell ◽  
R. R. Pelosi ◽  
P. A. Rundell ◽  
M. Cohen
Keyword(s):  
Immunosorbent Assay ◽  
Citrus Tristeza Virus ◽  
Cross Protection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Experimental Area ◽  
Toxoptera Citricida ◽  
The Cross ◽  
Tristeza Virus ◽  
Citrus Tristeza

A 21-year-old replicated field planting of 84 ‘Ruby Red’ grapefruit trees cross-protected with three mild isolates of Citrus tristeza virus (CTV) was assessed for decline-inducing and non-decline-inducing isolates of the virus 5 years after the brown citrus aphid (BrCA) (Toxoptera citricida Kirkaldy) first was established in the experimental area. Prior to the introduction of the BrCA, the cross-protecting mild isolates had significantly reduced detectable infection with decline-inducing isolates of CTV for 16 years (average infection of 13% in cross-protected trees compared with 67% in unprotected trees). After the introduction of the BrCA, infections with decline-inducing CTV (measured by enzyme-linked immunosorbent assay) were 57, 81, and 71% for trees protected with three mild isolates, respectively, compared with 95% in unprotected trees. These results suggest that the introduction of BrCA accelerated the breakdown of cross-protection against decline-inducing isolates of CTV in grapefruit.

scholarly journals In Situ Immunoassay (ISIA) of Field Grapefruit Trees Inoculated with Mild Isolates of Citrus tristeza virus Indicates Mixed Infections with Severe Isolates

Plant Disease ◽  
2002 ◽  
Vol 86 (5) ◽  
pp. 458-461 ◽  
Author(s):  
Youjian Lin ◽  
Phyllis A. Rundell ◽  
Charles A. Powell
Keyword(s):  
Monoclonal Antibodies ◽  
Immunosorbent Assay ◽  
Citrus Tristeza Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mixed Infections ◽  
The Cross ◽  
Tristeza Virus ◽  
Mild Isolate ◽  
Citrus Tristeza

Ten grapefruit trees that had been inoculated with a mild isolate of Citrus tristeza virus (CTV) and maintained in the field for 18 years were found in a previous study to be declining and infected with severe isolates of CTV, or symptomless and infected with mild isolates of CTV, using enzyme-linked immunosorbent assay (ELISA). They were assayed with an in situ immunoassay (ISIA) procedure using monoclonal antibodies 17G11 (reacts with most Florida isolates of CTV) and MCA13 (reacts with severe, but not Florida mild isolates of CTV). All the grapefruit trees were 17G11 positive by ELISA and ISIA. The five trees that showed moderate decline symptoms were MCA13 positive by ELISA and ISIA. The five symptomless trees were MCA13 negative by ELISA. However, four of the five symptomless trees were MCA13 positive by ISIA, which showed that ISIA with MCA13 had greater sensitivity in detecting severe CTV isolates than ELISA. These results suggested that the cross-protected grapefruit trees, regardless of symptoms, were infected with both mild and severe isolates of CTV.

scholarly journals Cross-Protection of Grapefruit from Decline-Inducing Isolates of Citrus Tristeza Virus

Plant Disease ◽  
1999 ◽  
Vol 83 (11) ◽  
pp. 989-991 ◽  
Author(s):  
C. A. Powell ◽  
R. R. Pelosi ◽  
P. A. Rundell ◽  
E. Stover ◽  
M. Cohen
Keyword(s):  
Citrus Tristeza Virus ◽  
Natural Infection ◽  
Tree Height ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sour Orange ◽  
River Region ◽  
Indian River ◽  
Tristeza Virus ◽  
Better Than ◽  
Citrus Tristeza

The ability of three mild isolates of citrus tristeza virus (CTV) to prevent natural infection of 84 Ruby Red grapefruit on sour orange rootstock by aphid-transmitted, decline-inducing isolates of CTV was assessed by symptoms and verified by enzyme-linked immunosorbent assay (ELISA) after 16 years. Of 21 trees in each of four treatments protected by the DD 102 bb, Guettler HS, and DPI 1-12-5-X-E mild CTV isolates, 14, 10, and 14% were infected by severe isolates (MCA13 monoclonal antibody reactive) compared with 67% for unprotected control trees. The health of trees protected by the DD 102 bb CTV isolate was significantly better than that of unprotected control trees as measured by decline, tree ratings, and tree height. These data suggest that infection by certain mild isolates of CTV can cross-protect grapefruit trees on sour orange rootstock from decline-inducing isolates of CTV that are prevalent in the Indian River region of Florida.

scholarly journals PRODUCTION OF POLYCLONAL ANTIBODY TO THE COAT PROTEIN OF CITRUS TRISTEZA VIRUS IN CHICKEN EGGS

2016 ◽  
Vol 4 (1) ◽  
pp. 18
Author(s):  
Nurhadi Nurhadi ◽  
Kamaruzaman Sijam ◽  
Inon Sulaiman
Keyword(s):  
Coat Protein ◽  
Polyclonal Antibody ◽  
Citrus Tristeza Virus ◽  
Sodium Dodecyl ◽  
Virus Titer ◽  
Healthy Plant ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sds Page ◽  
Tristeza Virus ◽  
Citrus Tristeza

Citrus tristeza virus (CTV) is one of the most destructive diseases in many citrus growing areas of Indonesia. Effective strategies for controlling CTV depend on diagnostic procedure namely enzyme-linked immunosorbent assay (ELISA). Study aimed to purify the CTV antigen and produced its polyclonal antibody. Virion of the severe CTV isolate designated UPM/ T-002 was concentrated by polyethylene glycol (PEG) precipitation combined with low speed centrifugation. Semipurified antigen was further purified by sodium dodecyl sulphatepolyacrylamide gel electrophoresis (SDS-PAGE). The specific coat protein (CP) band of CTV with molecular weight of 25 kD was excised and eluted using elution buffer containing 0.25 M Tris-HCl pH 6.8 + 0.1% SDS, then used as antigen for injection into 6-month-old female of White Leghorn chicken. Results, showed than the specific polyclonal antibody raised against the 25-kDa CP had a titer of approximately 104, gave low background reaction with healthy plant sap and reacted specifically with CTV isolates. The reaction was equally strong for a severe, a moderate, a mild, and a symptomless isolate, suggesting a broad reaction range of this antibody toward different CTV isolates. Optimal virus titer can be obtained since virus loss during purification could be minimized and the highly purified antigen as an immunogen could be obtained by cutting out the CP band from SDS-PAGE gels. Large amount of highly titer of CTV antibody can be produced in chicken egg. The simplicity of the procedure makes it economically acceptable and technically adoptable because the antibody can be produced in basic laboratory.

scholarly journals PRODUCTION OF POLYCLONAL ANTIBODY TO THE COAT PROTEIN OF CITRUS TRISTEZA VIRUS IN CHICKEN EGGS

2016 ◽  
Vol 4 (1) ◽  
pp. 18
Author(s):  
Nurhadi Nurhadi ◽  
Kamaruzaman Sijam ◽  
Inon Sulaiman
Keyword(s):  
Coat Protein ◽  
Polyclonal Antibody ◽  
Citrus Tristeza Virus ◽  
Sodium Dodecyl ◽  
Virus Titer ◽  
Healthy Plant ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sds Page ◽  
Tristeza Virus ◽  
Citrus Tristeza

Citrus tristeza virus (CTV) is one of the most destructive diseases in many citrus growing areas of Indonesia. Effective strategies for controlling CTV depend on diagnostic procedure namely enzyme-linked immunosorbent assay (ELISA). Study aimed to purify the CTV antigen and produced its polyclonal antibody. Virion of the severe CTV isolate designated UPM/ T-002 was concentrated by polyethylene glycol (PEG) precipitation combined with low speed centrifugation. Semipurified antigen was further purified by sodium dodecyl sulphatepolyacrylamide gel electrophoresis (SDS-PAGE). The specific coat protein (CP) band of CTV with molecular weight of 25 kD was excised and eluted using elution buffer containing 0.25 M Tris-HCl pH 6.8 + 0.1% SDS, then used as antigen for injection into 6-month-old female of White Leghorn chicken. Results, showed than the specific polyclonal antibody raised against the 25-kDa CP had a titer of approximately 104, gave low background reaction with healthy plant sap and reacted specifically with CTV isolates. The reaction was equally strong for a severe, a moderate, a mild, and a symptomless isolate, suggesting a broad reaction range of this antibody toward different CTV isolates. Optimal virus titer can be obtained since virus loss during purification could be minimized and the highly purified antigen as an immunogen could be obtained by cutting out the CP band from SDS-PAGE gels. Large amount of highly titer of CTV antibody can be produced in chicken egg. The simplicity of the procedure makes it economically acceptable and technically adoptable because the antibody can be produced in basic laboratory.

scholarly journals Prevalence of Citrus Tristeza Virus in Florida Citrus Nurseries and Scion Groves

HortScience ◽  
2003 ◽  
Vol 38 (2) ◽  
pp. 244-245 ◽  
Author(s):  
Charles A. Powell ◽  
Robert R. Pelosi ◽  
Phyllis A. Rundell
Keyword(s):  
Citrus Tristeza Virus ◽  
Sweet Orange ◽  
Enzyme Linked Immunosorbent Assay ◽  
Nursery Trees ◽  
Florida Citrus ◽  
High Level ◽  
Individual Trees ◽  
Tristeza Virus ◽  
Composite Samples ◽  
Citrus Tristeza

None of 4190 sweet orange [Citrus sinensis (L.) Osb.] nursery trees of `Hamlin', `Midsweet', `Navel', and `Valencia' sampled from five Florida citrus nurseries were infected with a decline-inducing isolate of citrus tristeza virus (CTV) as judged by enzyme-linked immunosorbent assay (ELISA) using isolate-specific monoclonal antibodies. Two of the nurseries had a relatively high level of infection (37% to 100% of composite samples containing tissue from 10 trees) with nondecline-inducing (mild) isolates of CTV, depending on the cultivar. Three of the nurseries had a lower incidence of mild CTV (0% to 22% of 10 tree composite samples). No nursery was CTV-free. ELISA of individual trees used as budwood sources by the nurseries revealed that one tree out of 260 tested contained decline-inducing CTV, and 83 contained mild CTV. These results suggest that the budwood certification program adopted in 1997 has virtually eliminated decline-inducing CTV from commercial budwood supplies.

scholarly journals First Report of Citrus tristeza virus in Greece

Plant Disease ◽  
2002 ◽  
Vol 86 (3) ◽  
pp. 329-329 ◽  
Author(s):  
D. Dimou ◽  
J. Drossopoulou ◽  
E. Moschos ◽  
C. Varveri ◽  
F. Bem
Keyword(s):  
Citrus Tristeza Virus ◽  
Sweet Orange ◽  
Enzyme Linked Immunosorbent Assay ◽  
First Report ◽  
Vein Clearing ◽  
Carrizo Citrange ◽  
Sweet Lime ◽  
Tristeza Virus ◽  
Propagation Material ◽  
Citrus Tristeza

Large-scale surveys of Citrus spp. for the presence of Citrus tristeza virus (CTV) by the Ministry of Agriculture in Greece began in 1995. Over 26,000 trees have been tested by enzyme-linked immunosorbent assay and immunoprinting (2). In summer 2000, the first CTV-infected sweet orange cv. Lane Late tree grafted on CTV-tolerant Carrizo citrange was found in Argolis County, Peloponnese. This tree belonged to a batch of CAC propagation material (20 trees) illegally introduced from Spain in 1994, which was subsequently traced and found to be infected (45%). A follow-up search of trees grafted with the above material was undertaken in the two concerned regions (Argolis and Chania-Crete), and more than 3,500 trees have been removed. Extensive surveys continue to identify and destroy new infections. Few cases (15 of 16,800) of natural transmission to cultivars other than cv. Lane Late have been found. All of these have been close to the initially infected trees in the Argolis area. Surveys in spring 2001 were extended to certified propagation material of Clemenpons mandarin on Carrizo citrange imported from Spain, and 7 of 1,038 plants were infected (0.64%). The virus was successfully graft-transmitted to sweet orange cv. Madame Vinous and sweet lime seedlings, where it was identified by immunoprinting and reverse transcription-polymerase chain reaction (1). Mild vein clearing symptoms appeared on both indicators. Vein clearing on sweet lime was also accompanied by leaf cupping. To our knowledge, this is the first report of CTV in Greece. References: (1) A. Sambade et al. J. Virol. Methods 87:25, 2000. (2) C. Vela et al. J. Gen. Virol. 67:91, 1986.

Citrus Diseases Exotic to Florida: Citrus Tristeza Virus–Stem Pitting (CTV-SP)

EDIS ◽  
2022 ◽  
Vol 2022 (1) ◽  
Author(s):  
Amit Levy ◽  
Ozgur Batuman ◽  
Peggy Sieburth ◽  
Ajia Paolillo ◽  
Kuang-Ren Chung ◽  
...  
Keyword(s):  
Citrus Tristeza Virus ◽  
Viral Disease ◽  
Original Version ◽  
Educational Tool ◽  
Citrus Industry ◽  
Citrus Diseases ◽  
Florida Citrus ◽  
Tristeza Virus ◽  
Stem Pitting ◽  
Citrus Tristeza

This document is one in a series designed to provide important information on the causal agent, symptoms, and transmission of exotic citrus diseases. This information can be used as an educational tool to raise awareness about these diseases and for scouting and identification efforts. Disseminating information about the diseases to the citrus industry may prevent their introduction and spread in Florida. This document will focus on the exotic viral disease caused by isolates of citrus tristeza virus–stem pitting (CTV-SP). Original version: Chung, Kuang-Ren, and Ronald Brlansky. 2006. “Citrus Diseases Exotic to Florida: Citrus Tristeza Virus– Stem Pitting (CTV-SP)”. EDIS 2006 (7). https://doi.org/10.32473/edis-pp149-2006.

scholarly journals Increase and Patterns of Spread of Citrus Tristeza Virus Infections in Costa Rica and the Dominican Republic in the Presence of the Brown Citrus Aphid, Toxoptera citricida

1998 ◽  
Vol 88 (7) ◽  
pp. 621-636 ◽  
Author(s):  
T. R. Gottwald ◽  
S. M. Garnsey ◽  
J. Borbón
Keyword(s):  
Costa Rica ◽  
Dominican Republic ◽  
Spatial Autocorrelation ◽  
Citrus Tristeza Virus ◽  
Local Area ◽  
Enzyme Linked Immunosorbent Assay ◽  
Toxoptera Citricida ◽  
Area Of Influence ◽  
Tristeza Virus ◽  
Citrus Tristeza

Citrus tristeza virus (CTV) was monitored for 4 years by monoclonal antibody probes via enzyme-linked immunosorbent assay in four citrus orchards in northern Costa Rica and four orchards in the Dominican Republic following the introduction of the brown citrus aphid, Toxoptera citricida. The Gompertz nonlinear model was selected as the most appropriate in most cases to describe temporal increase of CTV. Ordinary runs analysis for association of CTV-positive trees failed to show a spatial relationship of virus status among immediately adjacent trees within or across rows. The beta-binomial index of dispersion for various quadrat sizes suggested aggregations of CTV-positive trees for all plots within the quadrat sizes tested. Spatial autocorrelation analysis of proximity patterns suggested that aggregation often existed among quadrats of various sizes up to four lag distances; however, significant lag positions discontinuous from the main proximity pattern were rare. Some asymmetry was also detected for some spatial autocorrelation proximity patterns. These results were interpreted to mean that, although CTV-positive trees did not often influence immediately adjacent trees, virus transmission was common within a local area of influence that extended two to eight trees in all directions. Where asymmetry was indicated, this area of influence was somewhat elliptical. The spatial and temporal analyses gave some insight into possible underlying processes of CTV spread in the presence of T. citricida and suggested CTV spread was predominantly to trees within a local area. Patterns of longer-distance spread were not detected within the confines of the plot sizes tested. Longer-distance spread probably exists, but may well be of a complexity beyond the detection ability of the spatial analysis methods employed, or perhaps is on a scale larger than the dimensions of the plots studied.

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