Compositions and methods for detecting hepatitis B virus
View Patent ↗Compositions, methods and kits for detecting viral nucleic acids. Targets that can be detected in accordance with the invention include HBV and/or HIV-1 and/or HCV nucleic acids. Particularly described are oligonucleotides that are useful as hybridization probes and amplification primers that facilitate detection of very low levels of HBV nucleic acids.
1. A composition of oligonucleotides for amplifying and detecting HBV nucleic acid, wherein said composition comprises first and second amplification oligomers configured to generate an HBV amplicon from an HBV nucleic acid present in a test sample, wherein:
(a) the first amplification oligomer consists of (i) a target hybridizing sequence consisting of SEQ ID NO:24 or SEQ ID NO:27, (ii), optionally, an upstream sequence that is not complementary to an HBV target sequence, and (iii) a 5′ promoter sequence for an RNA polymerase; and
(b) the second amplification oligomer consists of a target hybridizing sequence consisting of SEQ ID NO:15.
2. The composition of claim 1 , wherein the first amplification oligomer sequence consists of SEQ ID NO:41 or SEQ ID NO:44.
3. The composition of claim 1 , wherein the first amplification oligomer target hybridizing sequence consists of SEQ ID NO:24.
4. The composition of claim 3 , further comprising a third amplification oligomer consisting of a target hybridizing sequence consisting of SEQ ID NO:5, SEQ ID NO:8, or SEQ ID NO:9.
5. The composition of claim 4 , further comprising a fourth amplification oligomer consisting of (i) a target hybridizing sequence consisting of SEQ ID NO:17 or SEQ ID NO:20, (ii), optionally, an upstream sequence that is not complementary to an HBV target sequence, and (iii) a 5′ promoter sequence for an RNA polymerase.
6. The composition of claim 1 , further comprising a detectably labeled probe complementary to the HBV amplicon.
7. The composition of claim 6 , wherein the detectably labeled probe comprises a chemiluminescent compound, a fluorescent compound, a quencher compound, or a combination thereof.
8. The composition of claim 6 , wherein the detectably labeled probe is a molecular beacon.
9. The composition of claim 1 , wherein the composition is provided as a component of a kit for the detection of HBV nucleic acid.
10. The composition of claim 1 , wherein the composition is provided as a component of a kit for screening donated blood samples or plasma samples for the presence of HBV.
11. The composition of claim 10 , further comprising components for amplification and/or detection of HIV-1 nucleic acids, HCV nucleic acids, or both.
12. A method for screening donated blood or blood products for the presence of HBV, comprising the steps of:
a. contacting a test sample from donated blood or a blood product with
(i) an amplification oligomer consisting of (i) a target hybridizing sequence consisting of SEQ ID NO:24 or SEQ ID NO:27, (ii), optionally, an upstream sequence that is not complementary to an HBV target sequence, and (ii) a 5′ promoter sequence for an RNA polymerase; and
(ii) an amplification oligomer consisting of a target hybridizing sequence consisting of SEQ ID NO:15;
b. amplifying any HBV nucleic acid that is present in the test sample in an isothermal in vitro nucleic acid amplification reaction to generate an HBV amplicon; and
c. detecting the HBV amplicon, wherein detecting the presence of HBV in the test sample indicates that the blood product or the donated blood contains HBV nucleic acids,
wherein steps b. and c. are performed simultaneously.
13. The method of claim 12 , wherein step c. is performed using a detectably labeled probe complementary to the HBV amplicon.
14. A method for screening plasma for the presence of HBV, comprising the steps of:
a. contacting a test sample from plasma with
(i) an amplification oligomer consisting of (i) a target hybridizing sequence consisting of SEQ ID NO:24 or SEQ ID NO:27, (ii), optionally, an upstream sequence that is not complementary to an HBV target sequence, and (iii) a 5′ promoter sequence for an RNA polymerase; and
(ii) an amplification oligomer consisting of a target hybridizing sequence consisting of SEQ ID NO:15;
b. amplifying any HBV nucleic acid that is present in the test sample in an isothermal in vitro nucleic acid amplification reaction to generate an HBV amplicon; and
c. detecting the HBV amplicon, wherein detecting the presence of HBV in the test sample indicates that the plasma contains HBV nucleic acids,
wherein steps b. and c. are performed simultaneously.
15. The method of claim 14 , wherein step c. is performed using a detectably labeled probe complementary to the HBV amplicon.