Evaluation of enzyme immunoassays in the diagnosis of camel (<i>Camelus dromedarius</i>) trypanosomiasis: a preliminary investigation

Rae, P.; Thrusfield, Michael; Higgins, Anne Patti; Aitken, Colin; Jones, T.W.; Luckins, A.G.

doi:10.1017/s0950268800029976

Cited by 17 publications

(4 citation statements)

References 13 publications

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“…Recently, promising results have been obtained with indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (Elisa) methods. The IFAT has shown to be sensitive when employed in rabbits (Luckins et al 1978) and horses (Bakos & Bustamant 1982, Monzon 1987, Marques 1996 as well as the Elisatest in camels, horses, water buffaloes and cattle (Rae et al 1989, Payne et al 1991. The present work investigated clinical and parasitological features and the pattern of humoral immune response along the experimental infection with T. evansi in dogs.…”

Section: Diagnosis Of T Evansi Infection By Conventionalmentioning

confidence: 98%

Clinical, Parasitological and Immunological Aspects of Experimental Infection with Trypanosoma evansi in Dogs

Aquino¹,

Machado²,

Alessi³

et al. 1999

Mem. Inst. Oswaldo Cruz

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Section: Diagnosis Of T Evansi Infection By Conventionalmentioning

confidence: 98%

Clinical, Parasitological and Immunological Aspects of Experimental Infection with Trypanosoma evansi in Dogs

Aquino¹,

Machado²,

Alessi³

et al. 1999

Mem. Inst. Oswaldo Cruz

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“…However, serum antibodies are not detectable in early T. e ansi infections, and antibody responses can persist after chemotherapy [18]. To improve the detection of current infections, antigen-detection ELISAs (AgELISAs), which detect specific antigenic determinants of T. e ansi and other trypanosomes in the circulation, have been developed [19][20][21]. Ag-ELISAs have been used to monitor tsetse-transmitted trypanosomosis control programmes in Uganda [22] and Zanzibar [23] but, to date, T. e ansi Ag-ELISAs have not been used widely in Southeast Asia, where parasitological tests are still commonly used.…”

Section: Introductionmentioning

confidence: 99%

The occurrence of Trypanosoma evansi in buffaloes in Indonesia, estimated using various diagnostic tests

Davison

Thrusfield

Husein³

et al. 2000

Epidemiol. Infect.

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SUMMARYThe prevalence and incidence of Trypanosoma e ansi infections in village buffaloes in Central Java were estimated using parasitological tests, two antigen-detection ELISAs (2G6 Ag-ELISA and Tr7 Ag-ELISA), an antibody-detection ELISA (IgG ELISA) and a card agglutination test (CATT). Of 2387 village buffaloes tested in five districts, 4 % (95 % confidence interval [CI] : 3 %, 5 %) were positive with the microhaematocrit test (MHCT), 58 % (95 % CI : 56 %, 60 %) were positive with the 2G6 Ag-ELISA and 70 % (95 % CI : 68 %, 72 %) were positive with the Tr7 Ag-ELISA. An increasing prevalence with age was found and the proportion of positive buffaloes was highest in the over 84 months-old age-group (68 %) with the 2G6 Ag-ELISA and in the 37-60 months-old age-group (78 %) with the Tr7 Ag-ELISA. Parasitaemic buffaloes were found in more than half of the villages visited. Corrected village-specific prevalence values obtained with the two Ag-ELISAs ranged from 0 % to over 100 %, and prevalence differed significantly (P 0n0001) between villages in four of the five districts. Overall, 10 % of buffaloes tested in markets were found to be parasitaemic and 39, 56 and 47 % were found positive with the 2G6 Ag-ELISA, IgG ELISA and CATT, respectively. Incidence rates varied according to the test used and ranged from 0n22 (95 % CI : 0n09, 0n44) to 0n44 (95 % CI : 0n24, 0n76), per animal-year at risk, in two villages. The results highlight the importance of using validated diagnostic tests to obtain accurate estimates of prevalence and incidence. These parameters are needed, for example in mathematical models, for the development and evaluation of different control strategies for T. e ansi infections in buffaloes.

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“…Unexpectedly, protein A peroxidase reacted positively with some negative samples recording non-specific reaction ranged from 25% to 41%. This observation could be attributed to non-specific reaction of protein A with IgGs [ 28 ].…”

Section: Discussionmentioning

confidence: 99%

Preparation of goat and rabbit anti-camel immunoglobulin G whole molecule labeled with horseradish peroxidase

et al. 2017

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Aim:As the labeled anti-camel immunoglobulins (Igs) with enzymes for enzyme-linked immunosorbent assay (ELISA) are unavailable in the Egyptian market, the present investigation was directed for developing local labeled anti-camel IgG with horseradish peroxidase (HRP) to save hard curacy.Materials and Methods:For purification of camel IgG whole molecule, camel sera was preliminary precipitated with 50% saturated ammonium sulfate and dialyzed against 15 mM phosphate-buffered saline pH 7.2 then concentrated. This preparation was further purified by protein A sepharose affinity column chromatography. The purity of the eluted camel IgG was tested by sodium dodecyl sulfate polyacrylamide gel electrophoresi. Anti-camel IgG was prepared by immunization of goats and rabbits separately, with purified camel IgG. The anti-camel IgG was purified by protein A sepharose affinity column chromatography. Whole molecule anti-camel IgG was conjugated with HRP using glutraldehyde based assay. Sensitivity and specificity of prepared conjugated secondary antibodies were detected using positive and negative camel serum samples reacted with different antigens in ELISA, respectively. The potency of prepared conjugated antibodies was evaluated compared with protein A HRP. The stability of the conjugate at −20°C during 1 year was assessed by ELISA.Results:The electrophoretic profile of camel IgG showed four bands of molecular weight 63, 52, 40 and 33 kDa. The recorded sensitivity and specificity of the product are 100%. Its potency is also 100% compared to 58-75% of commercial protein A HRP. The conjugates are stable for 1 year at −20°C as proved by ELISA.Conclusion:Collectively, this study introduces goat and rabbit anti-camel IgG whole molecules with simple, inexpensive method, with 100% sensitivity, 100% specificity and stability up to 1 year at −20°C. The important facet of the current study is saving hard curacy. Future investigations are necessary for preparation of IgG subclasses.

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Evaluation of enzyme immunoassays in the diagnosis of camel (Camelus dromedarius) trypanosomiasis: a preliminary investigation

Cited by 17 publications

References 13 publications

Clinical, Parasitological and Immunological Aspects of Experimental Infection with Trypanosoma evansi in Dogs

Clinical, Parasitological and Immunological Aspects of Experimental Infection with Trypanosoma evansi in Dogs

The occurrence of Trypanosoma evansi in buffaloes in Indonesia, estimated using various diagnostic tests

Preparation of goat and rabbit anti-camel immunoglobulin G whole molecule labeled with horseradish peroxidase

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