Dot enzyme-linked immunosorbent assay (ELISA) for the detection of Toxocara infection using a rat model

Abstract

Toxocariasis is a zoonotic disease usually caused by dog and cat roundworms, and Detection and diagnosis is difficult in paratenic and accidental hosts, including humans, as they cannot be detected through conventional methods such as fecal examination. Diagnosis therefore relies on immunological methods and molecular methods such as enzyme-linked immunosorbent assay (ELISA) and Western Blot, which are both time-consuming and requires sophisticated equipment. In the Philippines, only a few studies are available on seroprevalence. Therefore, there is a need to adapt methods for serodiagnosis of infection in humans for the Philippine setting. A dot enzyme linked immunosorbent assay (dot-ELISA) was standardized using excretory-secretory antigens. Test sera were collected from laboratory rats (Sprague-Dawley strain) experimentally infected with embryonated eggs of and as well as rice field rats naturally infected with and sp. Optimum conditions used were 20 µg/ml antigen concentration and 1:10 serum dilution. The sensitivity, specificity, positive, and negative predictive values were 90% (95% CI 55.5-99.7%), 100% (95% CI 69.2-100.0%), 100% (95% CI 66.4-100%), and 90.9% (95% CI 58.7-99.8%), respectively. Dot-ELISA has the potential to be developed as a cheaper, simpler, and more practical method for detection of anti- antibodies on accidental hosts. This is a preliminary study conducted on experimental animals before optimization and standardization for human serum samples.

Plasma C-MYC level manifesting as an indicator in progression of breast cancer

“…The dot ELISA test was performed to detect the c-myc protein in the culture supernatant and cell lysate of MDA-MB-231 cell lines and nonbreast cells, using the method of Paller et al [26] with some modifications. The 2 μl cell culture supernatant was dotted onto nitrocellulose membranes (0.45 μm pore size).…”

Ocular toxocariasis: a neglected parasitic disease in Egypt

“…Several immunodiagnostic techniques have been used for the diagnosis of ocular toxocariasis, such as enzymelinked immunosorbent assay (ELISA) (Fillaux and Magnaval 2013;Jin et al 2013), Dot-ELISA test (Paller et al 2017), and western blot (Magnaval et al 2002;Jin et al 2013). The antigens used in these immunoassays include somatic extracts of adult worms, embryonated eggs, intact or sectioned larvae, and Toxocara excretory-secretory antigen (TES).…”

Human Toxocariosis: Is it not a Public Health Problem Yet?

“…There are several experiences checking human infection by sero diagnosis in different countries [7][8][9] with prevalence between 5% to 13% in United States and 37% in Nigeria. Several diagnostic methods have been used with different technologies as recombinant antigens 10,11 dot ELISA 12 or ELISA and PCR 13 And the association with allergy and asthma, [14][15][16] ocular disease 16,17 and presence in playgrounds [18][19][20] is cause of interest for some researchers.…”