High affinity probes for analysis of human papillomavirus expression
First Claim
1. A high affinity probe for expression analysis of an mRNA, comprising a nucleobase sequence complementary to a target sequence of an mRNA.
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Abstract
This invention is directed toward methods kits and compositions pertaining to the use of high affinity probes for diagnostic applications. The invention is further directed toward the use of the high affinity probes for the detection of clinically important human papillomavirus (HPV) strains. High affinity probes enable rapid diagnostic assays and supply sufficient hybridization specificity to discriminate closely related HPV strains. Examples of high-affinity probes are provided which are specifically directed towards the detection of the E6 and E7 regions, and splice variants thereof of HPV mRNA.
34 Citations
80 Claims
- 1. A high affinity probe for expression analysis of an mRNA, comprising a nucleobase sequence complementary to a target sequence of an mRNA.
- 2. A high affinity probe for expression analysis of a cancer marker, comprising a nucleobase sequence complementary to a target sequence of the cancer marker.
- 4. A high affinity probe for expression analysis of human papillomavirus, comprising a nucleobase sequence complementary to a target sequence of human papillomavirus mRNA.
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47. A method of assessing the risk of developing HPV-based disease or cancer, comprising:
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a) contacting a sample from a subject with one or more high affinity probes complementary to a target sequence of human papillomavirus mRNA, and b) determining the presence of bound probe.
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59. A method for selecting subjects for treatment for HPV-based disease, comprising
a) contacting a sample from a subject with one or more high affinity probes complementary to a target sequence of human papillomavirus mRNA, b) determining the presence of bound probe, and c) correlating the presence of papillomavirus mRNA with a need for treatment for HPV-based diseases.
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62. A method for monitoring the efficacy of an HPV-treatment, comprising
a) determining a pre-treatment level of HPV infection, b) administering an HPV infection treatment, and c) determining a post-treatment level of HPV infection after an initial period of treatment.
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65. A method for the expression analysis of human papillomavirus by in situ hybridization, comprising:
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a) contacting the sample with at least one high-affinity probe that is substantially complementary to a portion of a HPV mRNA. b) incubating the sample with the high affinity probe; and
c) detecting the fluorescence of the sample, wherein the level of fluorescence is indicative of the presence and/or amount of mRNA within individual cell of the sample. - View Dependent Claims (66, 67, 68, 69)
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70. A method of diagnosing or predicting an HPV-based disease in a subject, comprising:
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a) determining a level of HPV infection by contacting a sample from a subject with one or more high affinity probes complementary to a target sequence of human papillomavirus mRNA, and determining the presence of bound probe;
b) comparing the level, to a standard level; and
c) correlating a modulated level in the cell from the subject with an indication of an HPV-based disease. - View Dependent Claims (71, 72, 73, 74)
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- 75. A kit for expression analysis of human papillomavirus comprising one or more high affinity probes and instructions for use in an in-situ hybridization assay.
- 79. A kit for the assessing the stage of HPV-based diseases, comprising one or more high affinity probes for expression analysis of human papillomavirus, wherein the proved comprises a nucleobase sequence complementary to a target sequence of human papillomavirus mRNA.
Specification