Compositions and Methods for Detecting Nucleic Acids of Epstein-Barr Virus
AssigneeGen-Probe Incorporated
InventorsAnkur SHAH, Jian Yu FUNG
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Abstract
Disclosed are compositions, methods, and kits that can be used to Epstein-Barr virus (EBV) in a sample undergoing testing. Nucleic acids of EBV can be isolated, amplified and detected with specificity, and without interference from non-EBV organisms. In some embodiments, nucleic acids used for amplification are isolated from human blood, or blood products. Nucleic acid isolation, amplification and detection steps can all be carried out using an automated instrument.
Claims59
A reaction mixture for determining the presence or absence of Epstein-Barr virus (EBV) in a sample, said reaction mixture comprising:
- a detection probe oligomer for detecting EBV nucleic acids,
- wherein the detection probe oligomer is up to 30 nucleotides in length and comprises the base sequence of SEQ ID NO:22 or the complement thereof, allowing for substitution of RNA and DNA equivalent bases;
- and
- a pair of amplification oligomers,
- wherein a first amplification oligomer of the pair comprises 18-25 contiguous bases of SEQ ID NO:3, allowing for substitution of RNA and DNA equivalent bases, and
- wherein a second amplification oligomer of the pair comprises 18-25 contiguous bases of SEQ ID NO:4, allowing for substitution of RNA and DNA equivalent bases.
A method of determining the presence or absence of Epstein-Barr virus (EBV) in a sample, said method comprising the steps of:
- (a) contacting a sample to be tested for the presence of EBV with an oligomer combination that comprises,
- a first amplification oligomer that comprises 18-25 contiguous bases of SEQ ID NO:3, allowing for substitution of RNA and DNA equivalent bases,
- a second amplification oligomer that comprises 18-25 contiguous bases of SEQ ID NO:4, allowing for substitution of RNA and DNA equivalent bases, and
- a detection probe oligomer of up to 30 nucleotides in length that comprises the base sequence of SEQ ID NO:22 or the complement thereof, allowing for substitution of RNA and DNA equivalent bases;
- (b) performing an in vitro nucleic acid amplification reaction using the oligomer combination, wherein any EBV target nucleic acid, if present in the sample, is a template for generating an amplification product;
- and
- (c) detecting, with the detection probe oligomer, the presence or absence of the amplification product, thereby determining the presence or absence of EBV in the sample.
A kit of reagents, comprising in one or more vials:
- a detection probe oligomer for detecting Epstein-Barr virus (EBV) nucleic acids,
- wherein the detection probe oligomer is up to 30 nucleotides in length and comprises the base sequence of SEQ ID NO:22 or the complement thereof, allowing for substitution of RNA and DNA equivalent bases;
- and
- a pair of amplification oligomers,
- wherein a first amplification oligomer of the pair comprises 18-25 contiguous bases of SEQ ID NO:3, allowing for substitution of RNA and DNA equivalent bases, and
- wherein a second amplification oligomer of the pair comprises 18-25 contiguous bases of SEQ ID NO:4, allowing for substitution of RNA and DNA equivalent bases.
A detection probe oligomer for detecting Epstein-Barr virus (EBV) nucleic acids, wherein the detection probe oligomer is up to 30 nucleotides in length and comprises the base sequence of SEQ ID NO:22 or the complement thereof, allowing for substitution of RNA and DNA equivalent bases.
Dependencies are inferred from the claim text.