PROBES FOR DETECTION OF HUMAN PARVOVIRUS NUCLEIC ACID
Nucleic acid oligomers specific for human parvovirus genomic DNA are disclosed. An assay for amplifying and detecting human parvovirus genotypes 1, 2 and 3 nucleic acid in biological specimens is disclosed. Compositions for amplifying and detecting the presence of human parvovirus genotypes 1, 2 and 3 genomic DNA in human biological specimens are disclosed.
1 . A detection probe oligomer comprising:
(a) a target-binding sequence that is SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:32, or SEQ ID NO:36; and
(b) a detectable label.
2 . The detection probe oligomer of claim 1 , wherein the detectable label is a chemiluminescent or fluorescent label.
3 . The detection probe oligomer of claim 1 , wherein the detectable label is a homogeneous detectable label.
4 . The detection probe oligomer of claim 3 , wherein the homogeneous detectable label is a chemiluminescent label.
5 . The detection probe oligomer of claim 4 , wherein the chemiluminescent label is an acridinium ester (AE) compound.
6 . The detection probe oligomer of claim 1 , wherein the detection probe oligomer has at least one 2′-methoxy linkage in the nucleic acid backbone.
7 . A probe reagent comprising:
(a) a detection probe oligomer comprising (i) a target-binding sequence that is SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:32, or SEQ ID NO:36, and (ii) a detectable label;
(b) succinate
(c) lithium chloride;
(d) lithium lauryl sulfate;
(e) at least one anti-coagulant selected from the group consisting of EDTA and EGTA; and
(f) ethanol.
8 . The probe reagent of claim 7 , wherein the lithium lauryl sulfate is present in the reagent at a concentration of 2% (w/v).
9 . The probe reagent of claim 7 , wherein the EDTA is present in the reagent at a concentration of 20 mM.
10 . The probe reagent of claim 7 , wherein the EGTA is present in the reagent at a concentration of 20 mM.
11 . The probe reagent of claim 7 , wherein the succinate is present in the reagent at a concentration of 100 mM.
12 . The probe reagent of claim 7 , wherein the detection probe oligomer has at least one 2′-methoxy linkage in the nucleic acid backbone.
13 . A method for the detection of an amplified human parvovirus nucleic acid comprising the steps of:
(a) providing an amplified human parvovirus nucleic acid comprising a target specific sequence that contains SEQ ID NO:88, or comprising a target specific sequence amplifiable with a primer consisting of SEQ ID NO:13 and a promoter primer consisting of SEQ ID NO:23;
(b) hybridizing the amplified human parvovirus nucleic acid with a detection probe oligomer, wherein the detection probe oligomer comprises (i) a target-binding sequence that is SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:32, or SEQ ID NO:36, and (ii) a detectable label; and
(c) detecting the label of the hybridized detection probe oligomer, thereby detecting the amplified human parvovirus nucleic acid.
14 . The method of claim 13 , wherein the detectable label is a chemiluminescent or fluorescent label.
15 . The method probe oligomer of claim 13 , wherein the detectable label is a homogeneous detectable label.
16 . The method oligomer of claim 15 , wherein the homogeneous detectable label is a chemiluminescent label.
17 . The method oligomer of claim 16 , wherein the chemiluminescent label is an acridinium ester (AE) compound.
18 . The method of claim 13 , wherein the detection probe oligomer has at least one 2′-methoxy linkage in the nucleic acid backbone.
19 . The method of claim 13 , further comprising the step of performing an isothermal amplification reaction on a sample suspected of containing human parvovirus to generate the amplified human parvovirus nucleic acid.
20 . The method of claim 19 , wherein the isothermal amplification reaction is a transcription-mediated amplification reaction.