DOT-ELISA for detection of anti-Cysticercus cellulosae antibodies in human cerebrospinal fluid using a new solid-phase (resin-treated polyester fabrics): preliminary report

Abstract

A dot enzyme-linked immunosorbent assay (DOT-ELISA) was developed to detect specific antibodies in cerebrospinal fluid (CSF) for human neurocysticercosis immunodiagnosis, with Cysticercus cellulosae antigen dotted on a new solid-phase. This was represented by sheets of a synthetic polyester fabric impregnated with a polymerized resin (N-methylol-acrylamide). A very stable preparation was thus obtained, the antigen being covalently bound by cross-linking with free N-methylol groups on the resin. Since robust, no special care was necessary for handling the solid-phase. The test could be performed at room-temperature. From 30 CSF samples assayed, 14 were positive, from a group of 15 cases of neurocysticercosis, with titers from 1 to 128; 15 other samples, from normals or other neurological diseases, were all negative. Test characteristics seem to indicate it as adequate for epidemiological surveys. A more detailed study on sensitivity, specificity, reproducibility and the use in serum samples is being conducted.

Dot-ELISA for the diagnosis of neurocysticercosis

“…Dot-ELISA has also been employed in NCC for the detection of cysticercus antibodies in the serum as well as in the CSF 9,36,37 . A preliminary study conducted on CSF samples for the presence of specific cysticercus antibodies, with the cysticercus antigen dotted on a new solid phase of synthetic polyester fabric impregnated with a polymerized resin, revealed, 14 out of 15 CSF samples of NCC to have titres ranging from one to 128 36 .…”

“…However, the test such as Western blot or plate ELISA although are specific and sensitive, have the disadvantages of being the tests that require expensive equipment, technical expertise, hence difficult to adapt in the field or in the not-so well equipped laboratories. Several authors have hence used dot-enzyme linked immunosorbent assay (Dot-ELISA), a simplified ELISA technique for detection of specific cysticercus antibodies in serum and CSF samples 9,36,37 . The aim of the present study was to standardize and evaluate dot-Enzyme linked immunosorbent assay (Dot-ELISA), a simple and rapid test for the detection of cysticercus antibodies in the serum for the diagnosis of neurocysticercosis in the less equipped laboratories or in the field situation.…”

Evaluación del método ELISA de punto para el diagnóstico de la cisticercosis humana y para estimar valores de prevalencia en una región endémica en Colombia.

“…En este estudio se estandarizó y se evaluó la prueba de EP utilizando parámetros semejantes a los reportados por otros investigadores en diferentes regiones del mundo (18)(19)(20)(21)(22). Cabe anotar que esta investigación registra el mayor número de muestras procesadas (933), ofrece comparación estadística entre diferentes muestras diagnósticas y evalúa la prueba no sólo a nivel de grupos de estudio predeterminados sino también a nivel de un grupo de población general.…”

“…Los diferentes estudios que han empleado el EP como prueba inmunodiagnóstica para cisticercosis han encontrado valores de sensibilidad y especificidad que varían aproximadamente entre 50% y 100% (18)(19)(20)(21)(22). Las diferencias de sensibilidad y especificidad reportadas frente a las obtenidas en este estudio pueden deberse a las diversas técnicas de preparación del antígeno, a las condiciones de ejecución propias de las pruebas, así como a la variabilidad existente en la composición antigénica de T. solium y cisticercos obtenidos de diferentes fuentes de una misma región o de regiones diferentes de un mismo país, hecho documentado por varios autores (23)(24)(25)(26).…”

A high degree of cross reactivity between the antigens of Taenia solium and Taenia crassiceps, potentially useful for the diagnosis of human neurocysticercosis (NCC)

“…A less sensitive serological test will select only high immunological responders. Although the same group initially reported high sensitivity in dot-ELISA, 7 others reported much lower sensitivity values. 8 This method of patient selection biases the estimates of sensitivity (over-estimating sensitivity) in the new test and antigens, and fails to demonstrate if similar responses to both antigen sources would hold when cases with low antibody responses are tested.…”