scholarly journals Sensitive and Specific Identification of Neospora caninum Infection of Cattle Based on Detection of Serum Antibodies to Recombinant Ncp29

2002 ◽  
Vol 9 (3) ◽  
pp. 611-615 ◽  
Author(s):  
Daniel K. Howe ◽  
Keliang Tang ◽  
Patricia A. Conrad ◽  
Karen Sverlow ◽  
J. P. Dubey ◽  
...  
Keyword(s):  
Neospora Caninum ◽  
Surface Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sensitive Assay ◽  
Serum Antibodies ◽  
Caninum Infection ◽  
Major Surface Protein ◽  
Major Surface ◽  
Negative Controls ◽  
Important Disease

ABSTRACT Neosporosis is an economically important disease of dairy cattle caused by the protozoan Neospora caninum. Diagnostic tests for neosporosis are complicated by the potential for cross-reaction of antibodies to antigens that are similar between N. caninum and closely related parasites Toxoplasma gondii and Sarcocystis cruzi. To provide a sensitive and specific assay for detecting antibodies to N. caninum in the serum of infected animals, we have investigated a recombinant form of the antigen known as Ncp29 (rNcp29), which is a major surface protein of the parasite. Ncp29 is encoded by a gene that is homologous to the SAG1 gene previously characterized from T. gondii. An enzyme-linked immunosorbent assay (ELISA) was used to screen animals for the presence of serum antibodies specific to rNcp29. The rNcp29 ELISA readily distinguished between cattle known to be infected with N. caninum (optical density [OD] > 1.2 at 1:500 or greater dilution) and negative controls (OD < 0.5 at 1:500). Additionally, sera from animals that were infected with T. gondii or S. cruzi were negative. The rNcp29 ELISA developed here provides a specific and sensitive assay for detecting neosporosis in cattle.

scholarly journals Determination of antigenic domain in GST fused major surface protein (Nc-p43) of Neospora caninum

2001 ◽  
Vol 39 (3) ◽  
pp. 241 ◽  
Author(s):  
Eui-Sun Son ◽  
Hye-Jin Ahn ◽  
Jae-Hoon Kim ◽  
Dae-Yong Kim ◽  
Ho-Woo Nam
Keyword(s):  
Neospora Caninum ◽  
Surface Protein ◽  
Major Surface Protein ◽  
Major Surface ◽  
Antigenic Domain

scholarly journals Detection of Cattle Naturally Infected with Anaplasma marginale in a Region of Endemicity by Nested PCR and a Competitive Enzyme-Linked Immunosorbent Assay Using Recombinant Major Surface Protein 5

1998 ◽  
Vol 36 (3) ◽  
pp. 777-782 ◽  
Author(s):  
Susana Torioni de Echaide ◽  
Donald P. Knowles ◽  
Travis C. McGuire ◽  
Guy H. Palmer ◽  
Carlos E. Suarez ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Immunosorbent Assay ◽  
Nested Pcr ◽  
Surface Protein ◽  
Anaplasma Marginale ◽  
Epidemiological Studies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Major Surface Protein ◽  
Persistent Infections ◽  
Major Surface

A competitive enzyme-linked immunosorbent assay using recombinant major surface protein 5 (rMSP5-cELISA) of Anaplasma marginale was validated in a naturally infected cattle herd in an area of eastern Oregon where A. marginale is endemic. The true positive and negative A. marginale infection status of 235 randomly selected cattle was determined by using a nested PCR (nPCR) coupled with msp5 sequence analysis and hybridization. Judgment of the reliability of the nPCR and hybridization for detection of persistent infections was based on three observations. First, the nPCR was able to detect as few as 30 infected erythrocytes per ml. Second, the nPCR was able to consistently detect low levels of rickettsemia in seven carrier cattle experimentally infected with A. marginale. Third, msp5sequence analysis showed >95% identity among 30 nPCR amplicons from cattle naturally infected with field strains of A. marginale. The nPCR and hybridization identified 151 infected and 84 uninfected cattle among the 235 animals tested. With a cutoff point of 28%, the rMSP5-cELISA showed a sensitivity of 96% and a specificity of 95%. These results indicate that the rMSP5-cELISA can sensitively and specifically detect cattle with naturally acquired persistent A. marginale infections and suggest that it is an excellent assay for epidemiological studies, eradication programs, and regulation of international cattle movement.

scholarly journals Serologic Cross-Reactivity between Anaplasma marginale and Anaplasma phagocytophilum

2005 ◽  
Vol 12 (10) ◽  
pp. 1177-1183 ◽  
Author(s):  
U. M. Dreher ◽  
J. de la Fuente ◽  
R. Hofmann-Lehmann ◽  
M. L. Meli ◽  
N. Pusterla ◽  
...  
Keyword(s):  
Animal Species ◽  
Surface Protein ◽  
Anaplasma Marginale ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antibody Assay ◽  
Serological Tests ◽  
Major Surface Protein ◽  
New Findings ◽  
Major Surface

ABSTRACT In the context of a serosurvey conducted on the Anaplasma marginale prevalence in Swiss cattle, we suspected that a serological cross-reactivity between A. marginale and A. phagocytophilum might exist. In the present study we demonstrate that cattle, sheep and horses experimentally infected with A. phagocytophilum not only develop antibodies to A. phagocytophilum (detected by immunofluorescent-antibody assay) but also to A. marginale (detected by a competitive enzyme-linked immunosorbent assay). Conversely, calves experimentally infected with A. marginale also developed antibodies to A. phagocytophilum using the same serological tests. The identity of 63% determined in silico within a 209-amino-acid sequence of major surface protein 5 of an isolate of A. marginale and one of A. phagocytophilum supported the observed immunological cross-reactivity. These observations have important consequences for the serotesting of both, A. marginale and A. phagocytophilum infection of several animal species. In view of these new findings, tests that have been considered specific for either infection must be interpreted carefully.

scholarly journals Expression and Immunogenicity of Recombinant Immunoreactive Surface Protein 2 of Anaplasma phagocytophilum

2012 ◽  
Vol 19 (6) ◽  
pp. 919-923 ◽  
Author(s):  
Qiang Yu ◽  
Chuang-fu Chen ◽  
Qiang Chen ◽  
Li-juan Zhang
Keyword(s):  
Recombinant Protein ◽  
Anaplasma Phagocytophilum ◽  
Surface Protein ◽  
Immunofluorescence Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Content Type ◽  
Major Surface Protein ◽  
Granulocytic Anaplasmosis ◽  
Major Surface ◽  
Diagnostic Reagent

ABSTRACTHuman granulocytic anaplasmosis (HGA), caused byAnaplasma phagocytophilum, is an emerging tick-borne zoonotic disease throughout the world. The first HGA cases in China were documented in 2008, and the greatest challenge posed by the disease is rapid and accurate diagnosis during the acute phage of illness. In this study, we successfully cloned and expressed anA. phagocytophilumimmunoreactive surface protein (major surface protein 2 [MSP2]) and demonstrated that this recombinant protein has natural immunogenicity by Western blotting and enzyme-linked immunosorbent assay (ELISA) using human HGA-positive sera and reference rabbit HGA-positive sera. The rabbit antisera against the recombinant protein also reacted actively with the natural antigen ofA. phagocytophilumby immunofluorescence assay (IFA). No cross-reaction was observed between the recombinant protein and rabbit antisera against 10 common members of the orderRickettsialesby ELISA when the sera were diluted more than 1:100. We concluded that the recombinant MSP2 protein exhibited excellent antigenicity and specificity, results that should lay the foundation for the development of a simple and rapid diagnostic reagent and a vaccination for anaplasmosis.

scholarly journals Evaluation of an Enzyme-Linked Immunosorbent Assay Using Recombinant Major Surface Protein 5 for Serological Diagnosis of Bovine Anaplasmosis in Venezuela

1998 ◽  
Vol 5 (2) ◽  
pp. 259-262 ◽  
Author(s):  
Armando Reyna-Bello ◽  
Axel Cloeckaert ◽  
Nieves Vizcaíno ◽  
Mary I. Gonzatti ◽  
Pedro M. Aso ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Serum Antibody ◽  
Surface Protein ◽  
Serological Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Major Surface Protein ◽  
Bovine Anaplasmosis ◽  
Major Surface ◽  
Positive Results

ABSTRACT An indirect enzyme-linked immunosorbent assay (ELISA) was developed for the serological diagnosis of bovine anaplasmosis with purified recombinant major surface protein 5 (MSP5) of Anaplasma marginale produced in Escherichia coli. Serum antibody responses against MSP5 were detected in calves experimentally infected with A. marginale as early as 21 days postinfection and reached maximum titers at 28 days postinfection. The MSP5 ELISA performed with serum samples taken from field cattle from different regions of Venezuela showed a seroprevalence of 47%, which seems to be in accordance with the reported epidemiological status of bovine anaplasmosis in Venezuela. Positive results obtained in the MSP5 ELISA were further confirmed by immunoblotting, with the recombinant MSP5 as the antigen. Thus, these results confirmed the importance of MSP5 as a suitable antigen for the serological diagnosis of bovine anaplasmosis.

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