Compositions and methods to detect
View Patent ↗Compositions are disclosed as nucleic acid sequences that may be used as amplification oligomers, including primers, capture probes for sample preparation, and detection probes specific for Candida albicans 26S rRNA sequences or DNA encoding 26S rRNA. Methods are disclosed for detecting the presence of C. albicans in samples by using the disclosed compositions in methods that include in vitro nucleic acid amplification of a 26S rRNA sequence or DNA encoding the 26S rRNA sequence to produce a detectable amplification product.
1. A method of detecting Candida albicans in a sample, comprising the steps of:
mixing a sample that contains a C. albicans target nucleic acid that is a 26S rRNA sequence or DNA encoding the 26S rRNA sequence with a first amplification oligomer of up to 100 nucleotides in length comprising a target binding region consisting of SEQ ID NO: 19 and a second amplification oligomer of up to 100 nucleotides in length comprising a target binding region consisting of SEQ ID NO: 24;
providing an enzyme with nucleic acid polymerase activity and nucleic acid precursors to make an amplification mixture that includes the first and second amplification oligomers and the C. albicans target nucleic acid;
elongating in vitro a 3′ end of at least one of the amplification oligomers hybridized to the C. albicans target nucleic acid by using the enzyme with nucleic acid polymerase activity and the C. albicans target nucleic acid as a template to produce an amplified product; and
detecting the amplified product by hybridizing the amplified product specifically to a detection probe oligomer of up to 100 nucleotides in length comprising a target binding sequence consisting of SEQ ID NO: 28 to indicate the presence Candida albicans in the sample.
2. The method of claim 1 , further comprising a sample processing step that captures the C. albicans target nucleic acid from the sample before the mixing step.
3. The method of claim 2 , wherein the sample processing step uses a capture probe oligomer that contains a target binding sequence consisting of SEQ ID NO: 35 or 36, wherein the target binding sequence is optionally covalently attached to a 3′ tail sequence.
4. A composition for detecting Candida albicans 26S rRNA sequence or DNA encoding the 26S rRNA sequence by using in vitro amplification, comprising a first amplification oligomer of up to 100 nucleotides in length comprising a target binding region consisting of SEQ ID NO: 19, a second amplification oligomer of up to 100 nucleotides in length comprising a target binding region consisting of SEQ ID NO: 24, and a detection probe oligomer of up to 100 nucleotides in length comprising a target binding region consisting of SEQ ID NO: 28.
5. The composition of claim 4 , further comprising at least one capture probe oligomer that contains a target binding region consisting of SEQ ID NO: 35 or 36, optionally with a 3′ tail sequence covalently attached to the target specific sequence.