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New methods of HPV identification *
Author(s) -
Kapranos Nikiforos C.
Publication year - 1992
Publication title -
journal of the european academy of dermatology and venereology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.655
H-Index - 107
eISSN - 1468-3083
pISSN - 0926-9959
DOI - 10.1111/j.1468-3083.1992.tb00672.x
Subject(s) - nucleic acid , dot blot , polymerase chain reaction , microbiology and biotechnology , southern blot , in situ hybridization , typing , nucleic acid thermodynamics , dna , hybridization probe , genomic dna , blot , biology , western blot , virology , gene , messenger rna , genetics , base sequence
Identification of human papilloma viruses (HPV) is of major clinical importance because certain types are strongly associated with neoplastic transformation, especially in the uterine cervix. These viruses are not reliably detected by serological tests or culture. The detection of viral capsid antigens by immunohistochemical techniques is of low sensitivity and cannot be used in typing of HPV. Molecular biology techniques based on nucleic acid hybridization are the most sensitive and specific for identification of HPV. Blot methods are applied on DNA extracted from cells or tissues. In the Southern blot technique, nucleic acid probes are bound with the electrophoresed DNA fragments whereas, in the dot blot test, the intact DNA is directly spotted onto filters and hybridized. Although the first method is considered the ‘gold standard’, the latter has a comparable sensitivity, is simpler and can also be used with non‐isotopic probes. In the filter in situ hybridization method, whole cells are denatured and hybridized on a membrane, and this results in high background and low specificity. In situ hybridization is applied directly to the cells and tissues and, although it is not as sensitive as blot methods, it permits comparison of pathological features and the retrospective study of archive material. Amplification of genomic sequences by polymerase chain reaction (PCR) makes possible their detection with non‐isotopic blot techniques. PCR, which is applicable on cells, fresh and fixed tissues, is an extremely valuable and sensitive technique for the identification of HPV in the clinical laboratory.