Development of an Indirect ELISA Based on a Recombinant Chimeric Protein for the Detection of Antibodies against Bovine Babesiosis

Ortiz, José Manuel Jaramillo; Montenegro, Valeria Noely; Fourniere, Sofía Ana María de la; Sarmiento, Néstor Fabián; Farber, Marisa Diana; Wilkowsky, Silvina Elizabeth

doi:10.3390/vetsci5010013

Cited by 12 publications

(6 citation statements)

References 19 publications

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“…However, cross reactions were observed when samples from B. bigemina infected cattle were analyzed, thus suggesting that using this assay is an alternative test to detect anti- Babesia sp. antibodies [74]. In general, ELISA has replaced the IFAT as the preferred diagnostic serological test in a diagnostic laboratory, particularly because of the ease and objectivity in interpretation, as well as for the capacity for automatization to process a higher number of samples in a working day.…”

Section: Indirect Immunologic Assaysmentioning

confidence: 99%

Diagnostic Tools for the Identification of Babesia sp. in Persistently Infected Cattle

2019

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Bovine babesiosis is a tick-borne disease of cattle caused by the protozoan parasites of the genus Babesia. Babesia bovis, Babesia bigemina and Babesia divergens are considered by International health authorities (OIE) as the principal species of Babesia that cause bovine babesiosis. Animals that recover from a babesial primo infection may remain as persistent carriers with no clinical signs of disease and can be the source of infection for ticks that are able to acquire Babesia parasites from infected cattle and to transmit Babesia parasites to susceptible cattle. Several procedures that have been developed for parasite detection and diagnosis of this infectious carrier state constitute the basis for this review: A brief description of the direct microscopic detection of Babesia-infected erytrocytes; PCR-based diagnostic assays, which are very sensitive particularly in detecting Babesia in carrier cattle; in-vitro culture methods, used to demonstrate presence of carrier infections of Babesia sp.; animal inoculation, particularly for B. divergens isolation are discussed. Alternatively, persistently infected animals can be tested for specific antibabesial antibodies by using indirect serological assays. Serological procedures are not necessarily consistent in identifying persistently infected animals and have the disadvantage of presenting with cross reactions between antibodies to Babesia sp.

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Section: Indirect Immunologic Assaysmentioning

confidence: 99%

Diagnostic Tools for the Identification of Babesia sp. in Persistently Infected Cattle

2019

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“…Various ELISA methods have been used to detect specific antibodies against Babesia species such as B. bovis , B. bigemina , B. divergens , B. caballi , B. canis , B. gibsoni , B. microti , and Babesia sp. Xinjiang ( Bose et al., 1990 ; Kappmeyer et al., 1999 ; Ikadai et al., 2000 ; Fukumoto et al., 2001 ; Boonchit et al., 2002 ; Goff et al., 2003 ; Goff et al., 2006 ; Goff et al., 2008 ; Niu et al., 2016 ; Chung et al., 2017 ; Ortiz et al., 2018 ). Recently, sandwich ELISA techniques based on antigen detection have been developed for the diagnosis of blood parasite infections ( De Arruda et al., 2004 ; Dondorp et al., 2005 ; Chen et al., 2008 ), including B. bovis and B. microti infections ( Montealegre et al., 1987 ; Luo et al., 2012 ; Thekkiniath et al., 2018 ).…”

Section: Discussionmentioning

confidence: 99%

Babesia ovis secreted antigen-1 is a diagnostic marker during the active Babesia ovis infections in sheep

Sevinc,

Zhou,

Cao

et al. 2023

Front. Cell. Infect. Microbiol.

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Ovine babesiosis caused by Babesia ovis is an economically significant disease. Recently, a few B. ovis-specific proteins, including recombinant B. ovis secreted antigen-1 (rBoSA1), have been identified. Immunological analyses revealed that rBoSA1 resides within the cytoplasm of infected erythrocytes and exhibits robust antigenic properties for detecting anti-B. ovis antibodies. This protein is released into the bloodstream during the parasite’s development. It would be possible to diagnose active infections by detecting this secretory protein. For this purpose, a rBoSA1-specific polyclonal antibody-based sandwich ELISA was optimized in this study. Blood samples taken from the naturally (n: 100) and experimentally (n: 15) infected sheep were analyzed for the presence of native BoSA1. The results showed that native BoSA1 was detectable in 98% of naturally infected animals. There was a positive correlation between parasitemia level in microscopy and protein density in sandwich ELISA. Experimentally infected animals showed positive reactions from the first or second day of inoculations. However, experimental infections carried out by Rhipicephalus bursa ticks revealed the native BoSA1 was detectable from the 7th day of tick attachment when the parasite began to be seen microscopically. Sandwich ELISA was sensitive enough to detect rBoSA1 protein at a 1.52 ng/ml concentration. Additionally, no serological cross-reactivity was observed between animals infected with various piroplasm species, including Babesia bovis, B. bigemina, B. caballi, B. canis, B. gibsoni, Theileria equi, and T. annulata. Taken collectively, the findings show that the rBoSA1-specific polyclonal antibody-based sandwich ELISA can be successfully used to diagnose clinical B. ovis infections in sheep at the early stage.

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“…The sensitivity and specificity rates reported in the current study are slightly lower compared with those described in a study using ELISA with a B. bovis chimerical multi-antigen (rMABbo-ELISA), which achieved sensitivity and specificity values of 95.9% and 94.3%, respectively. Such values may be due to the number of antigenic determinants containing the three B. bovis antigens that were used Merozoite Surface Antigen-2c (MSA-2c), Rhoptry-Associated Protein-1 (RAP-1), and heat shock protein 20), which would increase the probability for antibody recognition of B. bovis antigens [ 22 ]. Another study using more than one antigen (rBbSBP-1 + rBbSBP-4) in an indirect ELISA demonstrated the usefulness of this method for the detection of antibodies against B. bovis in cattle compared to results using a single antigen (rBbSBP-1 or rBbSBP-4) [ 37 ].…”

Section: Discussionmentioning

confidence: 99%

“…To avoid nonspecific antibody binding in the tests, all serum samples were subjected to a pre-adsorption process with E. coli cell lysate, as described previously [ 22 , 23 ]. Briefly, 200 µL of E. coli TOP10 (uninduced) cells cultured in LB medium and stored at −80 °C were resuspended in 1 mL of phosphate-buffered saline (PBS) containing 500 µL acid-washed glass beads (Sigma-Aldrich, St. Louis, MO, USA).…”

Section: Methodsmentioning

confidence: 99%

Comparative Study of Indirect Fluorescent Antibody, ELISA, and Immunochromatography Tests for Serological Diagnosis of Bovine Babesiosis Caused by Babesia bovis

Lira-Amaya

Martínez-García

Santamaria-Espinosa

et al. 2021

Animals

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The indirect fluorescent antibody test (IFAT) is the most frequently used test to conduct seroepidemiological studies so far, and it is regarded as the "gold standard" test for the serological diagnosis of bovine babesiosis. The aim of the present study was to compare the enzyme-linked immunosorbent assay (ELISA) and the rapid immunochromatography test (ICT) for use in the serological diagnosis of cattle exposed to B. bovis in Mexico. The evaluation of test performance was carried out with 30 positive and 30 negative reference sera. A total of 72 bovine sera samples collected from cattle in a region with endemic bovine babesiosis were analyzed by ELISA and ICT, and the results were compared with those of IFAT. Kappa value (k) was also calculated to determine the agreement between tests. The sensitivity and specificity of ELISA for detecting antibodies against B. bovis were 87% (26/30) and 80% (24/30), respectively. The sensitivity and specificity of ICT for detecting antibodies against B. bovis were 90% (27/30) and 83.3% (25/30), respectively. The overall concordance determined for ELISA and ICT was 94.4% (68/72) and 98.6% (71/72), respectively, when the results were compared with those of IFAT. ICT was more sensitive and specific in this comparative study, showing good strength of agreement (k = 0.79) with respect to IFAT. ICT combines a strip-based assay system that is fast, practical, and sensitive for detection of antibodies to B. bovis, which suggests that it could be applied in the field without requiring any laboratory equipment for its use and interpretation of test results.

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Development of an Indirect ELISA Based on a Recombinant Chimeric Protein for the Detection of Antibodies against Bovine Babesiosis

Cited by 12 publications

References 19 publications

Diagnostic Tools for the Identification of Babesia sp. in Persistently Infected Cattle

Diagnostic Tools for the Identification of Babesia sp. in Persistently Infected Cattle

Babesia ovis secreted antigen-1 is a diagnostic marker during the active Babesia ovis infections in sheep

Comparative Study of Indirect Fluorescent Antibody, ELISA, and Immunochromatography Tests for Serological Diagnosis of Bovine Babesiosis Caused by Babesia bovis

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