Novel test for rapid viral diagnosis: detection of adenovirus in nasopharyngeal mucus aspirates by means of nucleic-acid sandwich hybridisation

Abstract

Nucleic-acid sandwich hybridisation detected adenovirus in nasopharyngeal aspirates from children with acute respiratory infections within 20 h. The differentiation between subgroups of adenovirus (B and C) and the lack of false positives indicated the specificity of the method. The reference test was detection of an adenovirus protein (hexon) by means of radioimmunoassay. In nucleic-acid sandwich hybridisation the sample DNA mediates the binding of a labelled "probe" DNA fragment to a second DNA fragment bound on filter. The sample DNA itself is not fixed to a solid support, and the only pretreatment required is boiling in a detergent solution. The results indicate that the sandwich hybridisation is specific, quantitative, easy to do, and only marginally affected by the crude nature of the sample. It may therefore be a valuable addition to the range of rapid methods in microbial diagnosis.