Meyer, T. and Arndt, R. and Stockfleth, E. and Flammann, H. T. and Wolf, H. and Reischl, U. (1995) Strategy for typing human papillomaviruses by RFLP analysis of PCR products and subsequent hybridization with a generic probe. BIOTECHNIQUES, 19 (4). pp. 632-639. ISSN 0736-6205, 1940-9818
Full text not available from this repository.Abstract
Genital human papillomaviruses (HPV) were detected by PCR using L1 consensus primers MY09 and MY11. To determine the underlying HPV type(s), PCR products were subsequently analyzed employing a combination of restriction fragment length polymorphism (RFLP) and hybridization with a generic oligonucleotide probe that binds to a conserved region located close to the MY11-binding site within the PCR products. Using computer-assisted sequence analysis, the lengths of the corresponding BamHI, Ddel, HaeIII, HinfI and PstI restriction fragments hybridizing with the generic probe were calculated, revealing distinct patterns for each of the the 45 mucosal HPV types. This method is superior to RFLP analysis since it is not impaired by large amounts of restriction fragments resulting from nonspecific PCR products. Moreover; considering clinical specimens containing two or three different HPV types, direct sequencing of PCR products will be inconclusive, and the increased number of restriction fragments will complicate interpretation of RFLP patterns. Subsequent hybridization with the generic probe, however results in the appearance of, at most, 2 or 3 bands per restriction enzyme thus facilitates identification of the underlying HPV types.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | POLYMERASE CHAIN-REACTION; CERVICAL NEOPLASIA; PRIMERS; SCRAPES; DNA |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Immunologie Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 04 Jan 2024 09:17 |
| Last Modified: | 04 Jan 2024 09:17 |
| URI: | https://pred.uni-regensburg.de/id/eprint/52279 |
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