An evaluation of the dot-ELISA procedure as a diagnostic test in an area with a high prevalence of human Toxocara canis infection.

The aim of this work was to evaluate a dot-enzyme-linked immunosorbent assay (dot-ELISA) using excretory-secretory antigens from the larval stages of Toxocara canis for the diagnosis of toxocariasis. A secondary aim was to establish the optimal conditions for its use in an area with a high prevalence of human T. canis infection. The dot-ELISA test was standardised using different concentrations of the antigen fixed on nitrocellulose paper strips and increasing dilutions of the serum and conjugate. Both the dot-ELISA and standard ELISA methods were tested in parallel with the same batch of sera from controls and from individuals living in the problem area. The best results were obtained with 1.33 µg/mL of antigen, dilutions of 1/80 for the samples and controls and a dilution of 1/5,000 for the anti-human IgG-peroxidase conjugate. All steps of the procedure were performed at room temperature. The coincidence between ELISA and dot-ELISA was 85% and the kappa index was 0.72. The dot-ELISA test described here is rapid, easy to perform and does not require expensive equipment. Thus, this test is suitable for the serological diagnosis of human T. canis infection in field surveys and in the primary health care centres of endemic regions.

An evaluation of the dot-ELISA procedure as a diagnostic test in an area with a high prevalence of human Toxocara canis infection

The aim of this work was to evaluate a dot-enzyme-linked immunosorbent assay (dot-ELISA) using excretory-secretory antigens from the larval stages of Toxocara canis for the diagnosis of toxocariasis. A secondary aim was to establish the optimal conditions for its use in an area with a high prevalence of human T. canis infection. The dot-ELISA test was standardised using different concentrations of the antigen fixed on nitrocellulose paper strips and increasing dilutions of the serum and conjugate. Both the dot-ELISA and standard ELISA methods were tested in parallel with the same batch of sera from controls and from individuals living in the problem area. The best results were obtained with 1.33 µg/mL of antigen, dilutions of 1/80 for the samples and controls and a dilution of 1/5,000 for the anti-human IgG-peroxidase conjugate. All steps of the procedure were performed at room temperature. The coincidence between ELISA and dot-ELISA was 85 percent and the kappa index was 0.72. The dot-ELISA test described here is rapid, easy to perform and does not require expensive equipment. Thus, this test is suitable for the serological diagnosis of human T. canis infection in field surveys and in the primary health care centres of endemic regions.

Prueba de avidez de los anticuerpos IgG en la infección por Toxocara canis / IgG antibodies avidity test in Toxocara canis infection / O teste de avidez dos anticorpos IgG na infecção por Toxocara canis

A fin de evaluar la utilidad de la prueba de avidez de los anticuerpos IgG para el reconocimiento de la fase de infección de la toxocariosis en individuos con alto nivel de exposición al parásito, se estudiaron 142 niños asintomáticos de ambos sexos, de 1-6 años de edad, residentes en la ciudad de Resistencia (NE de Argentina), con eosinofilia > 10% y con exámenes coproparasitológicos negativos para helmintos. Se realizó la prueba de ELISA-IgG en todos los casos y en aquellos con títulos > 1/80 se determinó el índice de avidez de IgG. De los 142 niños estudiados, 82 (57,7%) resultaron positivos a la prueba de ELISA-IgG pero sólo 6 (7.3%) tuvieron Índice de avidez menor al 50%, cinco tenían entre 1 y 2 años de edad y el restante 5 años (mediana: 1,5 años). Cuatro tuvieron títulos moderados de IgG (entre 1/80 y 1/640) y 2 títulos altos (entre 1/1280 y 1/10240). Los otros 76 niños con índice de avidez mayor al 50% tenían entre 18 meses y 5,5 años de edad (mediana: 4 años), 44 tuvieron títulos serológicos moderados y 32, títulos altos. Los resultados hallados en este trabajo reafirman que la población de la ciudad de Resistencia presenta altos niveles de exposición a T. canis y que la determinación del índice de avidez de IgG resulta útil para el diagnóstico diferencial entre toxocariosis reciente o tardía, aunque es más efectiva para descartar una infección reciente que para confirmarla.

In order to evaluate the usefulness of the IgG avidity test to recognize the phase of T. canis human infection in individuals highly exposed to the parasite, 142 asymptomatic children of both sexes, 1-6 years old, living in Resistencia (Northeastern Argentina), with eosinophilia > 10% and without helminthic intestinal infestation were studied. ELISA-IgG test was performed in all cases, and in those with serological titers > 1/80 IgG avidity was also measured. Out of 142 children studied, 82 (57.7%) were positive to ELISA test, but only 6 (7.3%) had an IgG avidity index < 50%. Five of them were aged between 1 to 2 years and the other was 5 years old (median 1.5 years). Four had moderate serological titers (between 1/80 and 1/640) and two had high titers (between 1/1280 and 1/10240). The other 76 children with IgG avidity indexes > 50% were between 18 months and 5.5 years old (median 4 years), 44 had moderate serological titers and 32 had high titers. The results found in this study reaffirm that the population in Resistencia city is highly exposed to T. canis infection and that the IgG avidity test may be helpful in differential diagnosis of recent and past toxocariosis, though it would be more effective to rule out a recently acquired infection than to confirm it.

Com o fim de avaliar a utilidade da prova de avidez dos Acs IgG para o reconhecimento da fase de infecção de toxocaríase em indivíduos com alto nível de exposição ao parasita, foram estudadas 142 crianças assintomáticas de ambos os sexos de 1 a 6 anos de idade, residentes na cidade de Resistencia (noreste argentino) com eosinofilia > 10% e com exames coproparasitológicos negativos para helmintos. Foi realizada a prova Elisa-IgG em todos os casos e naqueles com títulos > 1/80 foi determinado o índice de avidez de IgG. Das 142 crianças estudadas, 82 (57,7%) resultaram positivas à prova de Elisa-IgG porém só 6 (7,3%) tiveram índice de avidez < a 50%, 5 tinham entre 1 e 2 anos de idade e as outras 5 anos (média: 1,5 anos). Quatro tiveram títulos moderados de IgG (entre 1/80 e 1/640) e 2 títulos altos (entre 1/1280 e 1/10240). As outras 76 crianças com índice de avidez maior a 50% tinham entre 18 meses e 5,5 anos de idade (média: 4 anos), 44 tiveram títulos sorológicos moderados e 32 títulos altos. Os resultados encontrados neste trabalho reafirmam que a população da cidade de Resistencia apresenta altos níveis de exposição à T. canis e que a determinação do índice de avidez de IgG resulta útil para o diagnostico diferencial entre toxocaríase recente ou tardia, embora seja mais efetiva para descartar uma infecção recente que para confirmá-la.