| 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 polymerase chain reactions (PCR TM ) or other nucleic amplification techniques to amplify copies of the target DNA or RNA (via reverse-transcriptase followed by PCR, Murakawa et al. 1988 DNA 7-287) to allow for detection. The method of the present invention combines the following four steps for the first time: DNA amplicon synthesis and amplification in a modified nucleic acid amplification technique (eg. PCR) using a target nucleic acid sequence as a template; amplicon transcription into RNA sequences with amplification thereof to generate a significantly increased number complementary RNA sequences (i.e., transcriptional enhancement (TE) products); capture of TE products using immobilized complementary DNA sequences; and immunochemical detection of the heteroduplex nucleic acid sequences thereby formed. This method extends standard nucleic acid amplification techniques to accommodate transcription of the PCR products into RNA sequences with amplification, to significantly magnify the signal-to-noise ratio of the diagnostic test This combination of steps results in a new assay with increased simplicity, speed and sensitivity compared to standard nucleic acid detection methods.
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