Edward D Huguenel

Rapid identification of microorganisms by nucleic acid hybridization after labeling the test sample.

A novel method for rapidly identifying microorganisms has been developed. This method employs a monoadduct-forming furocoumarin derivative, which can photochemically label nucleic acids. The labeled nucleic acid can, in turn, be hybridized simultaneously to a panel of immobilized probe DNAs arrayed as dots on a solid support such as nitrocellulose. This procedure offers several advantages over more conventional hybridization techniques in that sample nucleic acids can be photolabeled without substantial sample preparation and that identification can be achieved by a single, rapid hybridization reaction.

Detection of biotinylated nucleic acid hybrids by antibody-coated gold colloid

A simple and sensitive nonradioactive method for detecting biotinylated nucleic acid hybrids on nitrocellulose paper is described. The method employs anti-biotin antibody-coated gold colloid and silver enhancement. Comparative results indicate sensitivity similar to that of a commonly used enzymatic colorimetric detection method.

Nucleic Acid Hybridization: A Rapid Method for the Diagnosis of Infectious Diseases

A new method is described for the rapid identification of microorganisms in clinical samples by DNA-DNA hybridization. In this procedure, nucleic acids are labelled nonradioisotopically in the sample, which is then incubated with a panel of immobilized unlabelled DNAs from different organisms as probes. The sample labelling is a photochemical reaction between the nucleic acids in cell lysates and in isopsoralen derivative covalently linked to a biotin moiety for detection. Results are given of the application of the procedure to the identification of pathogens in clinical specimens of urine and cerebrospinal fluid.