A PCR based DNA hybridisation capture system for the detection of human cytomegalovirus. A comparative study with other identification methods

A simple, sensitive and specific colourimetric hybridisation method for the detection of HCMV DNA in clinical specimens is described. This method comb ines a PCR assay with a sensitive sandwich hybridisation assay. It relies o n the use of a specific capture probe linked covalently to polystyrene micr oplates and a specific polybiotinylated detection probe. Amplified DNA frag ments, sandwiched between these two probes, are detected by an enzymatic co lour reaction. This PCR-based colourimetric hybridisation method was compar ed with other known HCMV detection methods. Clinical specimens (n = 145, co rresponding to 106 patients) were tested by both a nested PCR assay and thi s colourimetric hybridisation method; and by either the culture method or t he pp65 antigenaemia test depending on the type of sample used. The results showed that the PCR-based hybridisation method has a specificity similar t o tissue culture, known as the conventional gold standard method, and could be used for the examination of the clinical specimens. (C) 1999 Elsevier S cience B.V. All rights reserved.