Procedurally Similar Competitive Immunoassay Systems for the Serodiagnosis of<i>Babesia Equi, Babesia Caballi, Trypanosoma Equiperdum</i>, and<i>Burkholderia Mallei</i>Infection in Horses

Katz, J. I.; Dewald, Richard; Nicholson, Judy M.

doi:10.1177/104063870001200108

Cited by 37 publications

(21 citation statements)

References 12 publications

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“…caballi and anti-B. equi horse sera may be necessary to establish the authenticity of the cross-reaction as well as to determine the appropriate cutoff titers for the practical diagnosis of B. equi and B. caballi in the ELISA (10,11,19).…”

Section: Discussionmentioning

confidence: 99%

Cloning of a Truncated Babesia equi Gene Encoding an 82-Kilodalton Protein and Its Potential Use in an Enzyme-Linked Immunosorbent Assay

et al. 2002

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To isolate Babesia equi genes encoding immunodominant proteins, a cDNA expression library prepared from B. equi mRNA was immunoscreened with B. equi-infected horse serum. Eighteen positive cDNA clones were obtained, and the clone that showed the strongest immunoreactivity, designated Be82, was further characterized. The Be82 gene consisted of 1,953 bp and contained a partial open reading frame lacking the 5-terminal sequence. As shown by Western blot analyses, immune sera from mice intraperitoneally injected with the Be82 gene product recognized the 82-and 52-kDa proteins of B. equi but not those of Babesia caballi. The glutathione S-transferase fusion protein expressed in Escherichia coli that was purified and used as the antigen in the enzyme-linked immunosorbent assay reacted specifically with B. equi-infected horse sera. These results suggest that the Be82 gene product is a potential diagnostic antigen candidate in the detection of B. equi infection in horses that will be useful both in the performance of epidemiological studies and in the granting of quarantine passes.Equine piroplasmosis is an economically important tickborne protozoan disease of horses reported worldwide. This disease is caused by Babesia equi and Babesia caballi (18). Babesia parasites destroy erythrocytes and induce fever, anemia, and icterus in infected horses (12). These parasites are usually detected in blood smears only during the acute stage of the infection, and animals that recover from the disease remain parasite carriers. These carriers, as well as those previously exposed or infected, can be identified serologically (6).Standard serological tests for babesiosis are the complement fixation test (CFT) and the indirect fluorescent antibody test (IFAT) (5), both of which require large amounts of parasite antigens, particularly in large-scale seroepidemiological surveys. Because of its low sensitivity and specificity, the CFT is unable to detect latent infection and fails to accurately discriminate between negative and carrier animals (24). With the IFAT, on the other hand, standardization is difficult, considering the subjectivity of the reader in assessing the results (3, 4). Weiland (25) has demonstrated the strong cross-reactivity of anti-B. caballi horse serum with the lysate of B. equi-infected erythrocytes using an enzyme-linked immunosorbent assay (ELISA). However, with Western blot analysis, Ikadai et al. (8) have noted an erratic or inconsistent cross-reactivity between the anti-B. equi serum and the lysate of B. caballi-infected erythrocytes. In view of the absence of equine piroplasmosis in Japan and the increasing number of horses imported to the country from places where the infection is endemic, the development of a highly specific and sensitive diagnostic system for B. equi is of urgent necessity.In this study, we isolated and determined the nucleotide sequence of a cDNA clone from a cDNA expression library prepared from B. equi mRNA and we attempted to develop and assess the efficacy of the ELISA system using the g...

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Section: Discussionmentioning

confidence: 99%

Cloning of a Truncated Babesia equi Gene Encoding an 82-Kilodalton Protein and Its Potential Use in an Enzyme-Linked Immunosorbent Assay

et al. 2002

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“…The Rose Bengal plate agglutination test and counterimmunoelectrophoresis technique reported earlier (13,19) either have technical shortcomings or have low specificity and/or sensitivity. Two competitive ELISAs with comparable sensitivity (98.9 and 98.6%) and specificity (100%) were described for glanders diagnosis (14,29). Due to the sophisticated techniques and high costs of mallein and monoclonal antibodies, these competitive ELISAs will not be available in routine laboratories.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Recombinant Proteins of Burkholderia mallei for Serodiagnosis of Glanders

Pal

Kumar

Malik

et al. 2012

Clin. Vaccine Immunol.

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Glanders is a contagious disease caused by the Gram-negative bacillus Burkholderia mallei. The number of equine glanders outbreaks has increased steadily during the last decade. The disease must be reported to the Office International des Epizooties, Paris, France. Glanders serodiagnosis is hampered by the considerable number of false positives and negatives of the internationally prescribed tests. The major problem leading to the low sensitivity and specificity of the complement fixation test (CFT) and enzyme-linked immunosorbent assay (ELISA) has been linked to the test antigens currently used, i.e., crude preparations of whole cells. False-positive results obtained from other diagnostic tests utilizing crude antigens lead to financial losses to animal owners, and false-negative results can turn a risk into a possible threat. In this study, we report on the identification of diagnostic targets using bioinformatics tools for serodiagnosis of glanders. The identified gene sequences were cloned and expressed as recombinant proteins. The purified recombinant proteins of B. mallei were used in an indirect ELISA format for serodiagnosis of glanders. Two recombinant proteins, 0375H and 0375TH, exhibited 100% sensitivity and specificity for glanders diagnosis. The proteins also did not cross-react with sera from patients with the closely related disease melioidosis. The results of this investigation highlight the potential of recombinant 0375H and 0375TH proteins in specific and sensitive diagnosis of glanders.

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“…Although IFAT is more sensitive than CFT and rarely renders false negative results FRIEDHOFF, 1986), standardization is difficult, considering the subjectivity of the reader in assessing the results (BÖSE et al, 1995;BRÜNING, 1996). ELISA is nowadays an alternative for increased specificity and sensitivity in the detection of acute and latent babesial infections and several procedures of ELISA using recombinant antigen or crude antigen have been standardized (BALDANI et al, 2007;KUMAR et al, 2003;HIRATA et al, 2003;XUAN et al, 2001;KATZ et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

Occurrence of Theileria equi in horses raised in the Jaboticabal microregion, São Paulo State, Brazil

Baldani

Nakaghi

Machado

2010

Rev. Bras. Parasitol. Vet.

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Blood and serum samples from 170 horses raised in the Jaboticabal microregion, São Paulo State, Brazil, were collected and tested by microscopic examination of blood smears, indirect fluorescent antibody test (IFAT) and nested polymerase chain reaction (nPCR) for Theileria equi infections. The association among the test results was verified by the McNemar test. During the examination of thin blood smears, parasites were detected in six (3.52%) horses. Anti-T. equi antibodies were detected in 100% sera samples, with titers ranging between 1:80 and 1:5120. The nPCR based on the T. equi merozoite antigen gene (EMA-1) allowed the visualization of specie-specific amplified product in 108 (63.53%) horses. All six samples judged positive microscopically were also positive for nPCR. Statistical analysis indicated general disagreement (p < 0.0001) between IFAT and nPCR; IFAT and blood smear; and nPCR and blood smear on the detection of parasite carriers. The results of the present study indicate that T. equi is widely spread among horses in the Jaboticabal microregion, Northeast region of São Paulo State, Brazil.

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Procedurally Similar Competitive Immunoassay Systems for the Serodiagnosis ofBabesia Equi, Babesia Caballi, Trypanosoma Equiperdum, andBurkholderia MalleiInfection in Horses

Cited by 37 publications

References 12 publications

Cloning of a Truncated Babesia equi Gene Encoding an 82-Kilodalton Protein and Its Potential Use in an Enzyme-Linked Immunosorbent Assay

Cloning of a Truncated Babesia equi Gene Encoding an 82-Kilodalton Protein and Its Potential Use in an Enzyme-Linked Immunosorbent Assay

Evaluation of Recombinant Proteins of Burkholderia mallei for Serodiagnosis of Glanders

Occurrence of Theileria equi in horses raised in the Jaboticabal microregion, São Paulo State, Brazil

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