Babesia gibsoni: Serodiagnosis of infection in dogs by an enzyme-linked immunosorbent assay with recombinant BgTRAP

Goo, Youn‐Kyoung; Jia, Honglin; Aboge, Gabriel Oluga; Terkawi, Mohamad Alaa; Kuriki, Ken; Nakamura, Chinatsu; Kumagai, Akiko; Zhou, Jinlin; Lee, Eung-goo; Nishikawa, Yoshifumi; Igarashi, Ikuo; Fujisaki, Kozo; Xu, Xuan

doi:10.1016/j.exppara.2007.11.010

Cited by 50 publications

(24 citation statements)

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“…Unexpectedly, rBbTRAP-T showed the lowest specificity and sensitivity with field serum samples, indicating that this antigen is not suitable for the development of a serological test. In a related study, rBgTRAP was noted as the best antigen for the detection of infection with B. gibsoni in canine sera (14,26). The discrepancies in performance might be due to differences in the host and immune responses as well as the low identical homology between the genes in the species.…”

Section: Discussionmentioning

confidence: 99%

“…Proteins derived from these organelles, coupled with the membrane component of parasites, are believed to have critical functions in parasite survival and growth (34). These proteins include (i) variable merozoite surface antigens (VMSAs) (4), which are believed to play key roles in the initial attachment of merozoites and sporozoites during their invasion (34), (ii) microneme and rhoptry proteins, including apical membrane antigen 1 (AMA-1), thrombospondin-related anonymous protein (TRAP), and rhoptry-associated protein 1 (RAP-1), which seem to be involved in the formation of a tight junction between merozoites and erythrocytes (RBC) (11), and (iii) spherical body proteins (SBPs), which have roles in stabilizing the environment after invasion and in aiding parasite growth (14,26). Importantly, they have been suggested as promising candidates for developing subunit vaccines or diagnostic antigens (34).…”

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confidence: 99%

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Spherical Body Protein 4 Is a New Serological Antigen for Global Detection of Babesia bovis Infection in Cattle

Terkawi

Huyên

Wibowo

et al. 2011

Clin Vaccine Immunol

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Five Babesia bovis recombinant proteins, including merozoite surface antigen 2c (BbMSA-2c), C-terminal rhoptry-associated protein 1 (BbRAP-1/CT), truncated thrombospondin-related anonymous protein (BbTRAP-T), spherical body protein 1 (BbSBP-1), and spherical body protein 4 (BbSBP-4), were evaluated as diagnostic antigens to detect the infection in cattle. The recombinant proteins were highly antigenic when tested with experimentally B. bovis-infected bovine serum in Western blot analysis. Furthermore, five antisera that had been raised against each of the recombinant proteins reacted specifically with the corresponding authentic protein, as determined in Western blot analysis. Next, enzyme-linked immunosorbent assays (ELISAs) using these recombinant proteins were evaluated for diagnostic use, and the sensitivity and specificity of each protein were demonstrated with a series of serum samples from experimentally B. bovis-infected cattle. Furthermore, a total of 669 field serum samples collected from cattle in regions of B. bovis endemicity in seven countries were tested with the ELISAs, and the results were compared to those of an indirect fluorescent antibody test (IFAT), as a reference. Among five recombinant antigens, recombinant BbSBP-4 (rBbSBP-4) had the highest concordance rate (85.3%) and kappa value (0.705), indicating its reliability in the detection of specific antibodies to B. bovis in cattle, even in different geographical regions. Overall, we have successfully developed an ELISA based on rBbSBP-4 as a new serological antigen for a practical and sensitive test which will be applicable for epidemiologic survey and control programs in the future.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Spherical Body Protein 4 Is a New Serological Antigen for Global Detection of Babesia bovis Infection in Cattle

Terkawi

Huyên

Wibowo

et al. 2011

Clin Vaccine Immunol

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“…The TRAP gene encodes one of the leading immunoreactive proteins in B. gibsoni [23]. Its protein product has been shown to be a promising vaccine candidate for protection against B. gibsoni infection [5,13]. Investigation into the extent of sequence variation in this antigen candidate may help elucidate the diversities of B. gibsoni strains and pave the way for development of effective serodiagnostic reagents as well as vaccines for canine babesiosis.…”

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confidence: 99%

Sequence and Phylogenetic Analysis of the Thrombospondin-Related Adhesive Protein (TRAP) Gene of Babesia gibsoni Isolates from Dogs in Taiwan

et al. 2010

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ABSTRACT. The genetic diversity of Babesia gibsoni strains worldwide is currently poorly defined. The aim of the present study was to characterize B. gibsoni strains in naturally infected dogs in Taiwan using a combination of polymerase chain reaction (PCR) and sequence analysis of both 18S rDNA and the gene encoding thrombospondin-related adhesive protein (TRAP). Genomic DNA was extracted from 29 parasitemic dogs, and the target genes were separately amplified, sequenced and aligned with corresponding sequences available in GenBank. All 18S rDNA sequences (1,262 bp) amplified from the Taiwanese isolates were identical to each other and had very high similarity (99.9-100%) with previously reported B. gibsoni sequences. These results provide the first molecular evidence showing infection of dogs with B. gibsoni from Taiwan. On the other hand, a phylogenetic analysis based on the deduced amino acid sequence of the TRAP gene demonstrated that the Taiwanese isolates were closely related to strains previously identified from Okinawa Island, Japan, but genetically distinct from strains found on Honshu in Japan and Jeju Island in South Korea. The divergence of TRAP among the geographically dispersed strains examined in this study and others supports the conclusion that this gene is useful for molecular genotyping of B. gibsoni strains.

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“…Immunoreactive recombinant proteins generated from parasite genes are valuable as diagnostic antigens and vaccine candidates (24)(25)(26)(27)(28)(29). Recombinant antigen-based serological assays have been widely and effectively used for the diagnosis of bovine, equine, and canine babesiosis (18)(19)(20)(21).…”

Section: Discussionmentioning

confidence: 99%

Identification and Expression of Babesia ovis Secreted Antigen 1 and Evaluation of Its Diagnostic Potential in an Enzyme-Linked Immunosorbent Assay

Sevinç

Cao

et al. 2015

J Clin Microbiol

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bIn order to identify immunoreactive proteins that are usable for the immunological diagnosis of Babesia ovis infections, a phage lambda cDNA expression library was constructed and screened using parasite-specific immune serum. Immunoscreening resulted in the identification of a full-length cDNA clone encoding a secreted protein designated Babesia ovis secreted antigen 1 (BoSA1). The full-length BoSA1 cDNA contained a 1,137-bp open reading frame that encoded a protein of 378 amino acids, with a signal peptide and 2 internal repeat domains. The theoretical molecular mass of the mature protein was 42.5 kDa. Recombinant BoSA1 (rBoSA1) protein was expressed in Escherichia coli strain DH5␣ cells as a glutathione S-transferase (GST) fusion protein and was purified by affinity chromatography. Purified rBoSA1 was tested for reactivity with sera from animals experimentally or naturally infected with B. ovis, in an indirect enzyme-linked immunosorbent assay (ELISA). The results showed that specific antibodies against rBoSA1 were detectable on days 7 and 8 of the experimental infection and were maintained during the sampling period. Additionally, 38 field sera taken from sheep naturally infected with B. ovis gave strong positive reactions in the ELISA between day 20 and day 30 of treatment. As a result, the identified recombinant BoSA1 protein seems to be a promising diagnostic antigen that is usable for the development of serological assays for the diagnosis of ovine babesiosis. This is the first report on the molecular cloning, expression, and potential use of a recombinant antigen for the diagnosis of ovine babesiosis. O vine babesiosis, caused by the tick-borne intraerythrocytic apicomplexan parasite Babesia ovis, is an acute disease characterized by clinical signs such as fever, hemolytic anemia, hemoglobinuria, and icterus in small ruminants. The disease is endemic in European, African, Asian, and Far Eastern countries (1-8). Babesia ovis is highly pathogenic and, in serious infections, causes pancytopenia. Untreated cases usually result in the death of sick animals, and even some treated animals may die as a result of heavy infection. Many recurrences also occur after the treatment period. Compensation for the abnormalities in the hematological profiles of acutely infected animals takes a long time after the treatment period (8). Currently, control of ovine babesiosis is based only on chemotherapy and limited tick control measures. In areas in which the status of B. ovis endemicity is unstable, an immunization program is needed (9). However, immunoprophylaxis is lacking for Babesia species in sheep.Currently, the diagnosis of ovine babesiosis includes microscopic examination of Giemsa-stained thin blood smears. Although intraerythrocytic parasites are apparent in clinical cases, it is very difficult to detect the organisms by microscopy in chronic infections, because of the low levels of parasitemia (10-12). The analysis of blood smears usually depends on the experience of the observer. The morphological changes in ...

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Babesia gibsoni: Serodiagnosis of infection in dogs by an enzyme-linked immunosorbent assay with recombinant BgTRAP

Cited by 50 publications

References 16 publications

Spherical Body Protein 4 Is a New Serological Antigen for Global Detection of Babesia bovis Infection in Cattle

Spherical Body Protein 4 Is a New Serological Antigen for Global Detection of Babesia bovis Infection in Cattle

Sequence and Phylogenetic Analysis of the Thrombospondin-Related Adhesive Protein (TRAP) Gene of Babesia gibsoni Isolates from Dogs in Taiwan

Identification and Expression of Babesia ovis Secreted Antigen 1 and Evaluation of Its Diagnostic Potential in an Enzyme-Linked Immunosorbent Assay

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