Methods and compositions for polynucleotide analysis using generic capture sequences

摘要

Methods, compositions and articles of manufacture for assaying a sample for an amplification product from a target polynucleotide are provided. An amplification reaction is used to produce the amplification product from the target polynucleotide so that it can be used to indirectly assay the sample for the target polynucleotide. A sample suspected of containing the target polynucleotide is contacted with first and second primers to amplify the target polynucleotide; the first primer comprises a tag sequence, the complement of which is formed on the opposite strand during amplification and is referred to as a capture sequence. That opposite strand is referred to as a second primer extension product or an amplification product, and comprises a label. A capture probe is provided that is conjugated to a substrate and can bind to the capture sequence to form an amplification product detection complex. Methods of detecting the amplification product thus produced are also provided, as are amplification product assay arrays, along with methods of forming the same. The methods are particularly useful in multiplex settings where a plurality of target polynucleotides are to be assayed. Kits comprising reagents for performing such methods are also provided.