PROBES FOR DETECTING CANDIDA GLABRATA
Disclosed are methods utilizing specific amplification of Candida sp. target nucleic acid for detecting the presence or absence of Candida sp. in a sample. Also disclosed are corresponding oligomers, including amplification oligomers, capture probes and detection probes, and combinations thereof, as well as corresponding reaction mixtures and kits.
1 . A detection probe for detecting a Candida ( C. ) glabrata target nucleic acid, said detection probe comprising
(a) a C. glabrata -specific detection probe target-hybridizing sequence consisting of a sequence selected from the group consisting of
the sequence of residues 1-17 of SEQ ID NO:60,
the sequence of residues 1-23 of SEQ ID NO:45,
the sequence of residues 1-17 of SEQ ID NO:18,
the sequence of residues 1-20 of SEQ ID NO:21,
the DNA equivalent or an RNA/DNA chimeric of any of the foregoing, and
the full complement of any of the foregoing; and
(b) at least one of
a label,
a substitution at the 2′ position of at least one ribose moiety, and
a blocking group at or near the 3′ end of the detection probe.
2 . The detection probe of claim 1 , wherein the detection probe has a sequence selected from the group consisting of the sequence of SEQ ID NO:60, SEQ ID NO:45, SEQ ID NO:18, SEQ ID NO:21, the DNA equivalent or an RNA/DNA chimeric of any of the foregoing, and the full complement of any of the foregoing.
3 . The detection probe of claim 1 , wherein the detection probe comprises the label.
4 . The detection probe of claim 3 , wherein the label is a chemiluminescent label or a fluorescent label.
5 . The detection probe of claim 3 , wherein the detection probe comprises a fluorescent label and a quencher.
6 . The detection probe of claim 5 , wherein the detection probe is a molecular torch, a molecular beacon, or a TaqMan detection probe.
7 . The detection probe of claim 1 , wherein the detection probe further comprises a non-target-hybridizing sequence.
8 . The detection probe of claim 7 , wherein the detection probe is a molecular torch or a molecular beacon.
9 . The detection probe of claim 1 , wherein the detection probe comprises the substitution at the 2′ position of at least one ribose moiety.
10 . The detection probe of claim 9 , wherein the substitution at the 2′ position of at least one ribose moiety is a 2′ methoxy substitution.
11 . The detection probe of claim 1 , wherein the detection probe comprises the blocking group at or near the 3′ end of the detection probe.
12 . The detection probe of claim 11 , wherein the blocking group is selected from the group consisting of an alkyl group, a non-nucleotide linker, an alkane-diol dideoxynucleotide residue, and cordycepin.
13 . A kit comprising the detection probe of claim 1 .
14 . The kit of claim 13 , further comprising at least one of
(i) an amplification oligomer combination for amplification of a C. glabrata nucleic acid target region, wherein the detection probe is capable of binding to said target region; and
(ii) a capture probe oligomer for purifying the C. glabrata target nucleic acid.
15 . The kit of claim 13 , further comprising a Candida -specific detection probe for detecting a Candida species (sp.) target nucleic acid from one or more of C. albicans, C. paripsolosis, C. dubliniensis , and C. tropicalis.
16 . The kit of claim 15 , further comprising at least one of
(i) an amplification oligomer combination for amplification of a Candida sp. nucleic acid target region, wherein said target region is from one or more of C. albicans, C. paripsolosis, C. dubliniensis , and C. tropicalis , and wherein the Candida -specific detection probe is capable of binding to said target region; and
(ii) a capture probe oligomer for purifying the Candida sp. target nucleic acid.
17 . A reaction mixture comprising the detection probe of claim 1 .
18 . The reaction mixture of claim 17 , further comprising at least one of
(i) an amplification oligomer combination for amplification of a C. glabrata nucleic acid target region, wherein the detection probe is capable of binding to said target region; and
(ii) a capture probe oligomer for purifying the C. glabrata target nucleic acid.
19 . The reaction mixture of claim 17 , further comprising a Candida -specific detection probe for detecting a Candida species (sp.) target nucleic acid from one or more of C. albicans, C. paripsolosis, C. dubliniensis , and C. tropicalis.
20 . The reaction mixture of claim 19 , further comprising at least one of
(i) an amplification oligomer combination for amplification of a Candida sp. nucleic acid target region, wherein said target region is from one or more of C. albicans, C. paripsolosis, C. dubliniensis , and C. tropicalis , and wherein the Candida -specific detection probe is capable of binding to said target region; and
(ii) a capture probe oligomer for purifying the Candida sp. target nucleic acid.