USE OF NUCLEIC ACID PROBES TO DETECT NUCLEOTIDE SEQUENCES OF INTEREST IN A SAMPLE

摘要

The invention relates to methods for the determination and detection of nucleic acids sequences in a sample. The nucleic acid may be RNA or DNA or both. The invention also relates to methods for the determination of the presence and species of various microorganisms in a sample. We have also identified a set of oligonucleotide nucleic acid sequences within the rRNAs of Gram-negative organisms that facilitates both the broad identification of Gram-negative organisms as a class when used as a pool or in combination, for example in a hybridization assay. This set of oligonucleotides may detect sequences that are indicative of the presence of organisms of the broad class of Gram-negative organisms while exhibiting little or no false identification of Gram- positive organisms, and fungi, or other microorganisms. The assay includes concurrent incubation with at least one nucleotide sequence of interest, at least one nucleic acid probe, a fluorosurfactant, and a nuclease. The assay may further be employed to detect the presence of bacteria, fungi, or other microorganisms by use of additional specific probes, or to detect and/or identify target nucleic acid sequences in a sample. Further, the invention also relates to methods of reducing non-specific binding and facilitating complex formation in a binding assay. The binding assay may be, but is not limited to, a nucleic acid hybridization assay or an immunoassay. The invention also relates to methods of detection that employ at least one target of interest, which may be a nucleotide sequence, at least one probe, which may be a nucleic acid probe and a nuclease.