scholarly journals Detection of Anaplasma antibodies in wildlife and domestic species in wildlife-livestock interface areas of Kenya by major surface protein 5 competitive inhibition enzyme-linked immunosorbent assay

2008 ◽  
Vol 75 (3) ◽  
Author(s):  
J.J.N. Ngeranwa ◽  
S.P. Shompole ◽  
E.H. Venter ◽  
A. Wambugu ◽  
J.E. Crafford ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Competitive Inhibition ◽  
Surface Membrane ◽  
Clinical Signs ◽  
Surface Protein ◽  
Domestic Animal ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Domestic Species ◽  
Major Surface

The seroprevalence of Anaplasma antibodies in wildlife (eland, blue wildebeest, kongoni, impala, Thomson's gazelle, Grant's gazelle, giraffe and plains zebra) and domestic animal (cattle, sheep and goat) populations was studied in wildlife / livestock interface areas of Kenya. Serum samples were analyzed by competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA), using a recombinant antigen (MSP-5) from Anaplasma marginale surface membrane. A monoclonal antibody, FC-16, was used as the primary antibody, while anti-mouse conjugated to horseradish peroxidase was used as the secondary antibody. The results indicate a high seroprevalence in both wildlife and livestock populations, in contrast to earlier reports from Kenya, which indicated a low seroprevalence. The differences are attributed to the accurate analytical method used (CI-ELISA), as compared with agglutination techniques, clinical signs and microscopy employed by the earlier workers.

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 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 Comparative examination of the serological response to bluetongue virus in diseased ruminants by competitive and double recognition enzime-linked immunosorbent assays

2018 ◽  
Vol 69 (3) ◽  
pp. 1088
Author(s):  
A. GAVRILOVIĆ ◽  
P. GAVRILOVIĆ ◽  
S. RADOJIČIĆ ◽  
D. KRNJAIĆ
Keyword(s):  
Blood Serum ◽  
Immunosorbent Assay ◽  
Clinical Signs ◽  
High Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Response ◽  
Rapid Diagnostics ◽  
Serum Samples ◽  
Perfect Agreement ◽  
First Time

Bluetongue (BT) is a viral non-contagious disease of ruminants which is transmitted by insects of the genus Culicoides. In recent years, BT has been a serious threat to livestock and to the economies of European countries. In Serbia the disease appeared for the first time in 2001, and after a 12 year period of freedom, it broke out again in 2014. Considering the actuality of this infectious disease, especially the need for prompt and rapid diagnostics, the aim of this paper was to determine the possibility of detecting the serological response in sheep and cattle with manifested clinical signs of the disease using two different methods: double recognition enzyme-linked immunosorbent assay (sELISA) and competitive enzyme-linked immunosorbent assay (cELISA). A total of 105 blood serum samples of cattle and sheep, which had exhibited clinical signs of BT during 2014, were taken for examination from a serum bank. Out of 74 blood serum samples of sheep and 31 blood serum samples of cattle, 52 samples of sheep and 18 samples of cattle tested positive using sELISA, while 50 samples of sheep and 18 samples of cattle gave positive reactions with cELISA. The results confirm the high sensitivity of sELISA which detected 4% more seropositive sheep in comparison with cELISA. Using Cohen’s kappa statistical analysis, almost perfect agreement was determined between the results (k>0,81) obtained by cELISA and sELISA.

scholarly journals  Toxoplasma gondii and Neospora caninum antibodies in goats in the Czech Republic

2012 ◽  
Vol 57 (No. 3) ◽  
pp. 111-114 ◽  
Author(s):  
E. Bartova ◽  
K. Sedlak
Keyword(s):  
Czech Republic ◽  
Toxoplasma Gondii ◽  
Immunosorbent Assay ◽  
Competitive Inhibition ◽  
Neospora Caninum ◽  
Fluorescent Antibody ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Antibodies ◽  
Serum Samples ◽  
The Czech Republic

Toxoplasma gondii is zoonotic protozoan parasite that causes infections in many vertebrate species. The present study determined the seroprevalence of T. gondii and N. caninum in goats from the Czech Republic. Serum samples were collected from 251 healthy adult goats in the Czech Republic during the years 2006 to 2009. Sera samples were tested for serum antibodies to Toxoplasma gondii by an enzyme-linked immunosorbent assay with cut off equal to or higher than 50% S/P. The same samples were tested for serum antibodies to Neospora caninum by a competitive-inhibition enzyme-linked immunosorbent assay with cut off equal to or higher than 30% inhibition; positive sera were confirmed by an indirect fluorescent antibody test with cut-off titre equal to or higher than 40. Sera positive in both tests were marked as positive. In total, 166 (66%) and 15 (6%) goat sera reacted positively for T. gondii and N. caninum antibodies, respectively. All sera positive for N. caninum antibodies were simultaneously positive for T. gondii antibodies. This is the first detection of N. caninum antibodies in goats in the Czech Republic. Our findings indicate that goats in the Czech Republic are frequently exposed to T. gondii, but less frequently to N. caninum.  

scholarly journals Development of a Highly Sensitive and Specific Enzyme-Linked Immunosorbent Assay Based on Recombinant Matrix Protein for Detection of Avian Pneumovirus Antibodies

2000 ◽  
Vol 38 (11) ◽  
pp. 4010-4014 ◽  
Author(s):  
Baldev R. Gulati ◽  
Kjerstin T. Cameron ◽  
Bruce S. Seal ◽  
Sagar M. Goyal ◽  
David A. Halvorson ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Matrix Protein ◽  
Clinical Signs ◽  
M Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Specific Test ◽  
Avian Pneumovirus ◽  
Highly Sensitive ◽  
The Matrix

The matrix (M) protein of avian pneumovirus (APV) was evaluated for its antigenicity and reliability in an enzyme-linked immunosorbent assay (ELISA) for diagnosis of APV infection, a newly emergent disease of turkeys in United States. Sera from APV-infected turkeys consistently contained antibodies to a 30-kDa protein (M protein). An ELISA based on recombinant M protein generated in Escherichia coli was compared with the routine APV ELISA that utilizes inactivated virus as antigen. Of 34 experimentally infected turkeys, 33 (97.1%) were positive by M protein ELISA whereas only 18 (52.9%) were positive by routine APV ELISA 28 days after infection. None of the serum samples from 41 uninfected experimental turkeys were positive by M protein ELISA. Of 184 field sera from turkey flocks suspected of having APV infection, 133 (72.3%) were positive by M protein ELISA whereas only 99 (53.8%) were positive by routine APV ELISA. Twelve serum samples, which were negative by M protein ELISA but positive by routine APV ELISA, were not reactive with either recombinant M protein or denatured purified APV proteins by Western analysis. This indicates that the samples had given false-positive results by routine APV ELISA. The M protein ELISA was over six times more sensitive than virus isolation (11.5%) in detecting infections from samples obtained from birds showing clinical signs of APV infection. Taken together, these results show that ELISA based on recombinant M protein is a highly sensitive and specific test for detecting antibodies to APV.

scholarly journals Detection of Equine Antibodies to Babesia caballi by Recombinant B. caballi Rhoptry-Associated Protein 1 in a Competitive-Inhibition Enzyme-Linked Immunosorbent Assay

1999 ◽  
Vol 37 (7) ◽  
pp. 2285-2290 ◽  
Author(s):  
Lowell S. Kappmeyer ◽  
Lance E. Perryman ◽  
Stephen A. Hines ◽  
Timothy V. Baszler ◽  
Jonathan B. Katz ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Competitive Inhibition ◽  
Immunofluorescence Assay ◽  
Cloning And Expression ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Gene Encoding ◽  
Peptide Epitope ◽  
Babesia Caballi ◽  
Serum Dilution

A competitive-inhibition enzyme-linked immunosorbent assay (cELISA) was developed for detection of equine antibodies specific forBabesia caballi. The assay used recombinant B. caballi rhoptry-associated protein 1 (RAP-1) and monoclonal antibody (MAb) 79/17.18.5, which is reactive with a peptide epitope of a native 60-kDa B. caballi antigen. The gene encoding the recombinant antigen was sequenced, and database analysis revealed that the gene product is a rhoptry-associated protein. Cloning and expression of a truncated copy of the gene demonstrated that MAb 79/17.18.5 reacts with the C-terminal repeat region of the protein. The cELISA was used to evaluate 302 equine serum samples previously tested for antibodies to B. caballi by a standardized complement fixation test (CFT). The results of cELISA and CFT were 73% concordant. Seventy-two of the 77 serum samples with discordant results were CFT negative and cELISA positive. Further evaluation of the serum samples with discordant results by indirect immunofluorescence assay (IFA) demonstrated that at a serum dilution of 1:200, 48 of the CFT-negative and cELISA-positive serum samples contained antibodies reactive with B. caballi RAP-1. Four of five CFT-positive and cELISA-negative serum samples contained antibodies reactive withB. caballi when they were tested by IFA. These data indicate that following infection with B. caballi, horses consistently produce antibody to the RAP-1 epitope defined by MAb 79/17.18.5, and when used in the cELISA format, recombinant RAP-1 is a useful antigen for the serologic detection of anti-B. caballi antibodies.

scholarly journals Competitive-Inhibition Enzyme-Linked Immunosorbent Assay for Detection of Serum Antibodies to Caprine Arthritis-Encephalitis Virus: Diagnostic Tool for Successful Eradication

2003 ◽  
Vol 10 (2) ◽  
pp. 267-271 ◽  
Author(s):  
Lynn M. Herrmann ◽  
William P. Cheevers ◽  
Travis C. McGuire ◽  
D. Scott Adams ◽  
Melinda M. Hutton ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Competitive Inhibition ◽  
False Negative ◽  
The United States ◽  
Encephalitis Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dairy Goats ◽  
Serum Antibodies ◽  
Serum Samples ◽  
Caprine Arthritis Encephalitis Virus

ABSTRACT A competitive-inhibition enzyme-linked immunosorbent assay (cELISA) was evaluated for the detection of serum antibodies to the surface envelope (SU) of caprine arthritis-encephalitis virus (CAEV) in goats. This assay utilized 96-well microtiter plates containing CAEV-63 SU captured by monoclonal antibody (MAb) F7-299 and measured the competitive displacement of horseradish peroxidase-conjugated MAb GPB 74A binding by undiluted goat sera (F. Özyörük, W. P. Cheevers, G. A. Hullinger, T. C. McGuire, M. Hutton, and D. P. Knowles, Clin. Diagn. Lab. Immunol. 8:44-51, 2001). Two hundred serum samples from goats in the United States were used to determine the sensitivity and specificity of cELISA based on the immunoprecipitation (IP) of [35S]methionine-labeled viral antigens as a standard of comparison. A positive cELISA was defined as >33.2% inhibition of MAb 74A binding based on 2 standard deviations above the mean percent inhibition of 140 IP-negative serum samples. At this cutoff value, there were 0 of 60 false-negative sera (100% sensitivity) and 5 of 140 false-positive sera (96.4% specificity). Additional studies utilized IP-monitored cELISA to establish a CAEV-free herd of 1,640 dairy goats.

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