Molecular diagnosis of infectious diseases by nucleic acid hybridization

Diagnostic microbiology has traditionally relied on the cultivation of an infecting agent in an in vitro system. However, the limitations of these methods have led to the development of alternative, molecular techniques which can detect specific microbial proteins or nucleic acids directly in body fluids. One such method, nucleic acid hybridization, has many attractive features including the potential for high sensitivity and specificity, the detection of a chemically stable analyte and the use of uniform reagents. Nucleic acid hybridization reactions have been used to detect a variety of microbial pathogens in assay formats using extracted nucleic acids or tissue sections. However, wider application of these techniques is limited by the fact that current methods employ radioisotopically labelled probes and cumbersome assay procedures. Solutions to these problems have been sought by the development of non-radioactive hybridization techniques which utilize enzymatic detection methods and by the development of rapid hybridization assays performed in solution. Further improvements in labelling techniques, assay formats and enzymatic detection methods should allow for the wider application of nucleic acid hybridization reactions in diagnostic microbiology.