Development of an Immunochromatographic Test with Recombinant EMA-2 for the Rapid Detection of Antibodies against
            <i>Babesia equi</i>
            in Horses

Huang, Xiaohong; Xu, Xuan; Xu, Lei; Shou-fa, Zhang; Yokoyama, Naoki; Suzuki, Naoyoshi; Igarashi, Ikuo

doi:10.1128/jcm.42.1.359-361.2004

Cited by 24 publications

(13 citation statements)

References 7 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The correspondence of the BceICT with BcELISA, BeELISA, BcICT, and BeICT were 91.8%, 95.9%, 98.2%, and 98.2%, respectively (Tables 1, 2, and 3). These results for B. equi infection were very comparable with those in previous studies (6).…”

Section: Discussionsupporting

confidence: 82%

“…Therefore, this test is more practical to use in the field than any other test. In our previous studies, the BcICT and BeICT were developed for the detection of antibodies to B. caballi (unpublished data) and B. equi (6). Both of the tests showed results that were comparable with those of ELISAs.…”

Section: Discussionmentioning

confidence: 66%

“…In our previous studies, ELISAs for the serodiagnoses of B. caballi and B. equi infections demonstrated many advantages, such as higher sensitivity and specificity, lower cost of materials, and greater objectivity in the determination of results (5,8), over the complement fixation test, indirect immunofluorescent antibody test, and competitive-inhibition ELISA. Compared with ELISA, however, the ICT is relatively simple, can be performed quickly, and has the listed advantages of ELISA (6).…”

mentioning

confidence: 99%

“…Various serodiagnostic tests have been developed for the disease, such as the complement fixation test (1,11,12), the indirect immunofluorescent antibody test (1,11,12), the enzyme-linked immunosorbent assay (ELISA) (1,3,5,7,8,9,10,13,14), the competitive-inhibition ELISA (9), and the immunochromatographic test (ICT) (6). In our previous studies, ELISAs for the serodiagnoses of B. caballi and B. equi infections demonstrated many advantages, such as higher sensitivity and specificity, lower cost of materials, and greater objectivity in the determination of results (5,8), over the complement fixation test, indirect immunofluorescent antibody test, and competitive-inhibition ELISA.…”

mentioning

confidence: 99%

See 3 more Smart Citations

Immunochromatographic Test for Simultaneous Serodiagnosis of Babesia caballi and B. equi Infections in Horses

Huang

Xuan

Verdida

et al. 2006

Clin Vaccine Immunol

Self Cite

View full text Add to dashboard Cite

An immunochromatographic test for the simultaneous detection of Babesia caballi-and B. equi-specific antibodies (BceICT) was developed using a recombinant B. caballi 48-kDa rhoptry protein (rBc48) and a recombinant truncated B. equi merozoite antigen 2 (rEMA-2t). An evaluation of the ability of the BceICT to detect antibodies in sera from uninfected horses and experimentally infected horses showed high sensitivities and specificities of 83.3% (10/12 sera) and 92.9% (52/56 sera), respectively, for the anti-B. caballi antibody and 94.1% (16/17 sera) and 88.2% (45/51 sera), respectively, for the anti-B. equi antibody. Results from the detection of antibodies in field-collected sera indicated that the BceICT results corresponded with those of enzyme-linked immunosorbent assays (ELISA), showing 91.8% correspondence (67/73 sera) for B. caballi and 95.9% correspondence (70/73 sera) for B. equi, and that the BceICT results also corresponded with the ICT for B. caballi and for B. equi, both of which were 98.2% (55/56 sera). The comparable results of the ICT and ELISA and the simplicity and rapidity of the performance of the ICT suggest that the BceICT would be a feasible test for the simultaneous serodiagnosis of both agents of equine babesiosis in the field.Equine piroplasmosis, caused by Babesia caballi and Babesia equi, is an important protozoan disease worldwide from both veterinary and economic viewpoints (2). Various serodiagnostic tests have been developed for the disease, such as the complement fixation test (1,11,12), the indirect immunofluorescent antibody test (1,11,12), the enzyme-linked immunosorbent assay (ELISA) (1,3,5,7,8,9,10,13,14), the competitive-inhibition ELISA (9), and the immunochromatographic test (ICT) (6). In our previous studies, ELISAs for the serodiagnoses of B. caballi and B. equi infections demonstrated many advantages, such as higher sensitivity and specificity, lower cost of materials, and greater objectivity in the determination of results (5, 8), over the complement fixation test, indirect immunofluorescent antibody test, and competitive-inhibition ELISA. Compared with ELISA, however, the ICT is relatively simple, can be performed quickly, and has the listed advantages of ELISA (6).Babesia caballi and B. equi have overlapping geographical distributions (4). In such areas, an individual horse may be infected by both species. Therefore, a test capable of detecting the antibodies induced by both types of parasites would be desirable. Here, we report an ICT for the simultaneous detection of B. caballi-and B. equi-specific antibodies (BceICT) that uses the recombinant B. caballi 48-kDa rhoptry protein (rBc48) and the recombinant truncated B. equi merozoite antigen 2 (rEMA-2t) as antigens for the simultaneous serodiagnosis of infections caused by two Babesia spp. in horses. MATERIALS AND METHODSrEMA-2t. rEMA-2 was expressed in Escherichia coli as a fusion protein with glutathione S-transferase, as described previously (5). The fusion protein was purified using glutathione Sepharose 4B (Amers...

show abstract

Section: Discussionsupporting

confidence: 82%

Section: Discussionmentioning

confidence: 66%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Immunochromatographic Test for Simultaneous Serodiagnosis of Babesia caballi and B. equi Infections in Horses

Huang

Xuan

Verdida

et al. 2006

Clin Vaccine Immunol

Self Cite

View full text Add to dashboard Cite

show abstract

“…Immunochromatographic tests have also been developed for the diagnosis of many protozoan diseases, such as babesiosis (Huang et al 2004b;Kim et al 2007Kim et al , 2008, malaria (Mills et al 1999), cryptosporidiosis (Chan et al 2000), leishmaniasis (Reithinger et al 2002), coccidiosis (Liao et al 2005), toxoplasmosis (Huang et al Table 2 Detection of anti-TaSP-specific antibodies in a cattle experimentally infected with T. annulata in serum taken at weekly intervals postinfection using iELISA, cELISA, and Ta-LFD iELISA cELISA Ta (Houghton et al 2009). Regarding the serodiagnosis of infection with T. annulata, there have been several reports in the last years, including IFAT (Burridge and Kimber 1973;Dhar and Gautam 1977;Darghouth et al 1996), indirect ELISA (Gubbels et al 2000;Bakheit et al 2004), and competitive ELISA (Renneker et al 2008).…”

Section: Discussionmentioning

confidence: 99%

Development and laboratory evaluation of a lateral flow device (LFD) for the serodiagnosis of Theileria annulata infection

Abdo

Kristersson²,

Seitzer

et al. 2010

Parasitol Res

View full text Add to dashboard Cite

Several DNA-based and serological tests have been established for the detection of Theileria annulata infection, including polymerase chain reaction, reverse line blot and loop-mediated isothermal amplification, indirect enzyme-linked immunosorbent assay (ELISA), and competitive ELISA. In this study, we have applied knowledge from the development and application of a recombinant protein-based indirect ELISA and competitive ELISA to establish a rapid test for point-of-care diagnosis of T. annulata infection in the field to be used by the veterinarian. For the development of a lateral flow test, the recombinantly expressed T. annulata surface protein (TaSP) was applied as the test antigen and anti-TaSP antiserum as the control line. TaSP antigen conjugated to colloidal gold particles was used as the detection system for visualization at the test line for the binding of anti-TaSP antibody present in the serum of infected animals. The developed test specifically detected antibodies in the serum of animals experimentally infected with T. annulata and showed no cross-reactivity with serum from animals infected with other tested bovine pathogens (Trypanosoma brucei, Anaplasma marginale, Babesia bigemina, Babesia bovis, and Theileria parva). Testing of field samples was compared to results obtained by other serological tests, resulting in a sensitivity and specificity of 96.3% and 87.5% compared to indirect fluorescence antibody test, 98.7% and 81.8% compared to indirect ELISA, and 100% and 47.6% compared to competitive ELISA. In conclusion, a rapid test for the detection of T. annulata infection (T. annulata lateral flow device, Ta-LFD) has been developed, which is easy to perform, delivers results to be read by the naked eye within 10 min, and is suitable for the detection of infection in field samples.

show abstract

Comparison of indirect fluorescent antibody test (IFAT) and slide enzyme linked immunosorbent assay (SELISA) for diagnosis of Babesia bigemina infection in bovines

Singh

Mishra

Rao

et al. 2008

Trop Anim Health Prod

View full text Add to dashboard Cite

An indirect fluorescent antibody test (IFAT) and slide enzyme linked immunosorbent assay (SELISA) were standardized for the detection of antibodies specific to Babesia bigemina in experimentally infected bovine calves and subsequently used for the screening of naturally infected bovine and bubaline sera. In experimentally infected calves positive reactivity was detected in sera at the earliest on day 7 by both the tests. Serological studies for detection of B. bigemina specific antibodies in 180 cow and 120 buffalo serum samples procured from endemic zones of Uttar Pradesh and Punjab revealed 56.11% and 23.33% seropositivity, respectively, both by SELISA and IFAT. Variation in the reactivity pattern between these tests was found to be non significant. The sensitivity of SELISA was determined to be 94.85% whereas the specificity was 90.85% in comparison to IFAT. The agreement between the two tests by kappa statistics at 95% confidence interval revealed kappa- value of 0.853 that depicts almost a perfect degree of agreement. The findings employing experimental as well as test sera from cattle and buffalo from some of the tick infested zones of India suggested that SELISA could be a useful tool for seroprevalence studies on babesiosis, as the test is less cost intensive with high levels of sensitivity and specificity.

show abstract

Development of an Immunochromatographic Test with Recombinant EMA-2 for the Rapid Detection of Antibodies against Babesia equi in Horses

Cited by 24 publications

References 7 publications

Immunochromatographic Test for Simultaneous Serodiagnosis of Babesia caballi and B. equi Infections in Horses

Immunochromatographic Test for Simultaneous Serodiagnosis of Babesia caballi and B. equi Infections in Horses

Development and laboratory evaluation of a lateral flow device (LFD) for the serodiagnosis of Theileria annulata infection

Comparison of indirect fluorescent antibody test (IFAT) and slide enzyme linked immunosorbent assay (SELISA) for diagnosis of Babesia bigemina infection in bovines

Contact Info

Product

Resources

About