Development of a prototype immunochromatographic test for rapid diagnosis of respiratory adenovirus infection

Abstract

Human adenoviruses comprise an important group of etiologic agents that are responsible for various diseases in adults and children, such as respiratory, ocular, gastroenteric, and urinary infections. In immunocompromised and organ-transplanted individuals, these agents can cause generalized infections. Rapid diagnostic methods for detecting these infectious agents are not widely available. The aim of this work was to produce monoclonal and polyclonal anti-adenovirus antibodies to be used in a rapid diagnostic test for respiratory infections. Adenovirus hexons were satisfactorily purified by ultracentrifugation and chromatography. After virus purification, anti-hexon monoclonal antibodies were produced and characterized, following classical methods. Antibodies were specific for adenoviruses 2, 3, 5, and 41. The proposed immunochromatographic test was standardized using colloidal gold. The standardization of the rapid test was sufficient to detect adenovirus antigens (in nasopharyngeal lavage samples) with sensitivity of 100% and specificity of 85% when compared to direct immunofluorescence. The immunochromatographic assay prototype was sufficiently sensitive to detect B (3), C (2 and 5), and F (41) adenovirus samples. Although based on preliminary data, the test demonstrated the same performance as direct immunofluorescence, but with the advantage of being a point-of-care test. Further studies are still needed to confirm its effectiveness in clinical practice.

Keywords: Adenovirus; Immunochromatographic test; Monoclonal antibodies.

Fig. 1

Prototype of immunochromatographic test for detection of adenovirus. Test line (positive) and control line (validation).

Fig. 2

Indirect ELISA of purified monoclonal antibodies (individual and combined reactions against culture adenovirus prototypes 41, 3, 2, and 5).

Fig. 3

Prototype of immunochromatographic test. C: control line. T: test line. Detection of cultured adenovirus (prototypes) 2, 3, 5, and 41 was successful.