The Use of Enzyme-Linked Immunosorbent Assay for Detection of Citrus Tristeza Virus

1979 ◽  
Vol 69 (2) ◽  
pp. 190 ◽  
Author(s):  
M. Bar -Joseph
Keyword(s):  
Immunosorbent Assay ◽  
Citrus Tristeza Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
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.

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