METHOD FOR ENHANCING DETECTION ABILITY OF NUCLEIC ACID ASSAYS EMPLOYING NUCLEIC ACID SEQUENCE BASED AMPLIFICATION

摘要

This invention relates to a new and novel method for significantly enhancing the sensitivity of assays aimed at detecting the presence of target deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) sequences in a sample. Specifically, this method applies to techniques that employ nucleic acid sequence based amplications (NASBA?) or other nucleic amplification techniques to amplify copies of the target RNA or DNA (Sooknanan et al., Bio"Techniques, 17: 1077-1085, 1994) to allow for detection. The method of the present invention combines the following three steps for the first time: RNA amplification using NASBA; RNA:DNA hybrid formation; and immunochemical detection of heteroduplex nucleic acid sequences. This method extends the standard NASBA techniques to accommodate amplification of copies of the NASBA product, which, following binding to DNA sequences facilitates immunochemical detection of the RNA:DNA hybrids thereby formed. This combination of steps results in a new assay with increased simplicity, speed and sensitivity, including a greatly improved signal-to-noise ratio, over standard nucleic acid detection methods. This improved method will be extremely useful for performing analyses in the forensic, clinical, agricultural and biological fields.