Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Patent
1991-02-15
1994-02-01
Parr, Margaret
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
435 6, 435810, 436501, 436811, 536 231, 536 243, 536 2431, 536 2432, 536 3433, 935 3, 935 20, 935 77, 935 78, 935 88, C12Q 168, C07H 2104, G01N 3350
Patent
active
052831710
DESCRIPTION:
BRIEF SUMMARY
The present invention provides medical research and diagnostic methods for detecting and typing HPV. The method utilizes PCR, a DNA amplification technique widely used in the fields of molecular biology and genetic engineering. The method can also be used to generate information concerning previously unknown strains of HPV and consequently has applications in the field of virology.
Papillomaviruses have been linked to widespread, serious human diseases, especially carcinomas of the genital and oral mucosa. And although genital HPV infection is associated with cancer primarily in women, recent evidence suggests that HPV may play a role in the development of prostate cancer in men. Broker et al., 1986, Cancer Cells 4:17-36, review the molecular, cellular, and clinical aspects of the papillomaviruses and the relationship of HPVs to cancer. HPV types 6, 11, 16, 18, and 33 are known genital HPV types in the human population, and Broker et al., 1986, Cancer Cells 4:589-594, disclose that HPV types 6, 11, 16, 18, and 33 share significant homology at the DNA level, particularly at the L1 open reading frame.
Identification and typing of HPV is quite important, because different types of HPV pose different risks to the affected individuals. For instance, HPV16 and HPV18 have been more consistently identified in higher grades of cervical dysplasia and carcinoma than other HPV types. Webb et al., December 1987, J. Inf. Disease 156(6):912-919, report a method for detecting HPV DNA types that utilizes a reverse-blotting procedure. The procedure involved forming a membrane to which genomic DNA from four different HPV types was bound and then hybridizing labelled DNA from a bilogical sample to the DNA bound to the membrane. Caussey et al., February 1988, J. Clin. Microbiol, 26(2):236-243 describe similar HPV detection methods.
Shibata et al., January 1988, J. Exp. Med, 167:225-230, disclose the use of PCR to amplify and detect the presence of HPV16 and HPV18 DNA. U.S. Pat. Nos. 4,683,195 and 4,683,202 disclose PCR and the use of PCR to detect the presence or absence of nucleic acid sequence in a sample. European Patent Publication Nos. 229,701 and 269,445 disclose the use of PCR to amplify and detect DNA sequences associated with a wide variety of viruses, including the AIDS virus, HTLV I, and HTLV II.
Maitland et al., May 1988, Seventh International Papillomavirus Workshop, Abstract, p. 5, report the use of PCR to detect HPV16 in oral and cervical biopsies. In addition, Campione-Piccardo et al., May 1988, Seventh International Papillomavirus Workshop, Abstract, p. 19, report the use of a mixture of primers for the specific amplification by PCR of HPV sequences in types 1a, 5, 6a, 8, 11, 16, 18, and 33. A number of other researchers disclosed the use of PCR to amplify and detect HPV sequences at the Seventh International Papillomavirus Workshop.
Despite the use of PCR to amplify and detect HPV sequences, there still remains a need for a simple and rapid method for both detecting and typing HPV in a biological sample. The present invention provides a method that meets that need.
The present invention provides a method for detecting and typing HPV in a sample. The method comprises amplifying a sequence of HPV DNA present in the sample, determining if amplification has occurred, and then hybridizing an HPV type-specific probe to the amplified DNA. The invention also provides novel primers and probes for use in the method.
The present invention provides a method for detecting HPV in a sample, and typing the HPV if present, the method comprising: polymerization, and deoxynucleoside 5'-triphosphates under conditions such that an extension product of a consensus primer can be synthesized, wherein said consensus primers are a mixture of oligonucleotides that comprises at least a pair of primers sufficiently complementary to separate single strands of HPV DNA to hybridize thereto so that the extension product synthesized from one member of said pair, when separated from its complementary strand, can serve as a template for synthes
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Broker Thomas R.
Manos M. Michele
Ting Yi
Wolinsky Steven M.
Wright Deann K.
Gould George M.
Hoffmann-La Roche Inc.
Marschel Ardin H.
Parr Margaret
Sias Stacey R.
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