Next-Generation Sequencing-Based Genotyping Assay for Human Papilloma Virus (HPV)
AssigneesUniversity of Maryland, Baltimore, Office of Technology Transfer, National Institutes of Health
InventorsKevin CULLEN, Nicholas AMBULOS, Jennifer TROYER
Life of this patent
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Abstract
Disclosed herein are compositions and methods for detecting and diagnosing HPV in subjects. The method includes using Next Gen Sequencing for detecting HPV in a sample and determining the specific subtype of HPV infection based on the sequencing data.
Claims37
A method for detecting HPV nucleic acid in a biological sample of a subject suspected of having an HPV infection or an HPV associated disease, the method comprising:
- (a) isolating a nucleic acid sample from a biological sample obtained from the subject;
- (b) determining the nucleic acid sequence of the nucleic acid sample using a next generation (Next Gen) sequencing assay;
- and (c) determining the genotype of HPV in the sample,
- wherein the method of determining the nucleic acid sequence of the nucleic acid sample comprises:
- (i) contacting the nucleic acid sample with at least one forward PCR primer, (ii) contacting the nucleic acid sample with at least one reverse PCR primer, (iii) amplifying the HPV nucleic acid using PCR, and (vi) sequencing the amplified nucleic acid products using a Next Gen Sequencer, and
- wherein the at least one forward PCR primer comprises a nucleic acid sequence selected from the group comprising SEQ ID NO:
- 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9 and further comprises a nucleic acid sequence for use as a sequencing adaptor.
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A composition comprising at least one PCR primer, wherein the at least one PCR primer comprises a nucleic acid sequence selected from the group comprising SEQ ID NO:
- 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9 and further comprises a nucleic acid sequence for use as a sequencing adaptor.
A composition comprising at least one PCR primer wherein the at least one PCR primer comprises a nucleic acid sequence selected from the group comprising SEQ ID NO:
- 11, SEQ ID NO:12 and SEQ ID NO:13 and further comprises a nucleic acid sequence for use as a barcode and a nucleic acid sequence for use as a sequencing adaptor.
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Dependencies are inferred from the claim text.