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 of Anaplasma marginale Major Surface Protein 2 Variants in Persistently Infected Ticks

2001 ◽  
Vol 69 (8) ◽  
pp. 5151-5156 ◽  
Author(s):  
José de la Fuente ◽  
Katherine M. Kocan
Keyword(s):  
Immune Response ◽  
Salivary Glands ◽  
Antigenic Variation ◽  
Surface Protein ◽  
Anaplasma Marginale ◽  
Dermacentor Variabilis ◽  
Persistently Infected ◽  
Major Surface Protein ◽  
Persistent Infections ◽  
Major Surface

ABSTRACT Anaplasma marginale, an intraerythrocytic ehrlichial pathogen of cattle, establishes persistent infections in both vertebrate (cattle) and invertebrate (tick) hosts. The ability ofA. marginale to persist in cattle has been shown to be due, in part, to major surface protein 2 (MSP2) variants which are hypothesized to emerge in response to the bovine immune response. MSP2 antigenic variation has not been studied in persistently infected ticks. In this study we analyzed MSP2 in A. marginalepopulations from the salivary glands of male Dermacentor variabilis persistently infected with A. marginaleafter feeding successively on one susceptible bovine and three sheep. New MSP2 variants appeared in each A. marginale population, and sequence alignment of the MSP2 variants revealed multiple amino acid substitutions, insertions, and deletions. These results suggest that selection pressure on MSP2 occurred in tick salivary glands independent of the bovine immune response.

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.

scholarly journals Anaplasma marginale Major Surface Protein 2 CD4+-T-Cell Epitopes Are Evenly Distributed in Conserved and Hypervariable Regions (HVR), Whereas Linear B-Cell Epitopes Are Predominantly Located in the HVR

2004 ◽  
Vol 72 (12) ◽  
pp. 7360-7366 ◽  
Author(s):  
Jeffrey R. Abbott ◽  
Guy H. Palmer ◽  
Chris J. Howard ◽  
Jayne C. Hope ◽  
Wendy C. Brown
Keyword(s):  
T Cell ◽  
B Cell ◽  
Surface Protein ◽  
Anaplasma Marginale ◽  
T Cell Epitopes ◽  
B Cell Epitopes ◽  
Major Surface Protein ◽  
Hypervariable Regions ◽  
Major Surface ◽  
Linear B

ABSTRACT Organisms in the genus Anaplasma express an immunodominant major surface protein 2 (MSP2), composed of a central hypervariable region (HVR) flanked by highly conserved regions. Throughout Anaplasma marginale infection, recombination results in the sequential appearance of novel MSP2 variants and subsequent control of rickettsemia by the immune response, leading to persistent infection. To determine whether immune evasion and selection for variant organisms is associated with a predominant response against HVR epitopes, T-cell and linear B-cell epitopes were localized by measuring peripheral blood gamma interferon-secreting cells, proliferation, and antibody binding to 27 overlapping peptides spanning MSP2 in 16 cattle. Similar numbers of MSP2-specific CD4+ T-cell epitopes eliciting responses of similar magnitude were found in conserved and hypervariable regions. T-cell epitope clusters recognized by the majority of animals were identified in the HVR (amino acids [aa] 171 to 229) and conserved regions (aa 101 to 170 and 272 to 361). In contrast, linear B-cell epitopes were concentrated in the HVR, residing within hydrophilic sequences. The pattern of recognition of epitope clusters by T cells and of HVR epitopes by B cells is consistent with the influence of protein structure on epitope recognition.

scholarly journals Comparison between indirect enzyme-linked immunosorbent assays for Anaplasma marginale antibodies with recombinant major surface protein 5 and initial body antigens

2006 ◽  
Vol 101 (5) ◽  
pp. 511-516 ◽  
Author(s):  
Virgínia MG Silva ◽  
Flábio R Araújo ◽  
Claudio R Madruga ◽  
Cleber O Soares ◽  
Raul H Kessler ◽  
...  
Keyword(s):  
Surface Protein ◽  
Anaplasma Marginale ◽  
Major Surface Protein ◽  
Immunosorbent Assays ◽  
Major Surface

Phylogeography of New World isolates of Anaplasma marginale based on major surface protein sequences

2002 ◽  
Vol 88 (3) ◽  
pp. 275-285 ◽  
Author(s):  
José de la Fuente ◽  
Ronald A Van Den Bussche ◽  
Jose C Garcia-Garcia ◽  
Sergio D Rodrı́guez ◽  
Miguel A Garcı́a ◽  
...  
Keyword(s):  
New World ◽  
Protein Sequences ◽  
Surface Protein ◽  
Anaplasma Marginale ◽  
Major Surface Protein ◽  
Major Surface
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