Probes for detection of human parvovirus nucleic acid

Abstract

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.

We claim: 
     
         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.