Detection and quantification of one or more nucleotide sequence target analytes in a sample using spatially localized target analyte replication

摘要

A target analyte which may be found in a substance, such as a biological or environmental substance, is assayed by inoculating a media with a sample of the substance. The target analyte is a unique nucleotide sequence of the RNA or DNA of a suspect organism, or any nucleotide target, or a combination of nucleotide sequences thereof, which organism may be found in the substance. The media contains selected target analyte amplifiers which will result in the amplification of any target analytes which are present in the substance. The media may also contain one or more labeled analyte-specific materials (LASMs) which can migrate through the media. The nature of the media is such that it will support target analyte-copying but will not allow the target analyte to migrate extensively within the media. After the substance to be assayed is added to the media and any target analytes are amplified, the LASMs will migrate by diffusion to resultant target analyte units or colonies in the media, thereby creating intensely labeled localized areas in the media which can be visually or mechanically detected. Each of the intensely labeled areas in the media will be surrounded by low intensity label halos. The number of high intensity labeled sites in the media will be proportional to the concentration of target analytes in the substance. IMAGE