New approaches to the diagnosis of oral soft-tissue disease of viral origin.
Molecular biology is changing the face of diagnostic medicine, and infectious diseases of the oral soft tissues are among the targets of these advances in biotechnology. As an illustration of these concepts, a PCR-based detection and typing system for human papillomaviruses (HPVs) will be discussed. A single "consensus" set of oligomeric nucleotide primers can be used to amplify a 571- to 594-base-pair region of the E1 open reading frame of HPV 6, 11, 16, and 18. These HPV types are commonly associated with preneoplastic and cancerous lesions of the genital, respiratory, and digestive tracts. PCR amplification yields single bands of similar size for these viruses by agarose gel electrophoresis. Digestion of the resultant products by the restriction endonuclease AccI yields distinctive and reproducible banding patterns by polyacrylamide gel electrophoresis (with ethidium bromide) due to their internal sequence diversity. The system is sensitive; without radioisotopes, it can detect and type HPV 18 in as little as 100 pg of DNA from HeLa cells. We have used it to confirm HPV in fresh-frozen tumors. Computer sequence analysis can be used to modify the system for the detection of new HPV types as they are characterized. Other applications of molecular-biology-based detection systems for infectious diseases of the head and neck region will be discussed.
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Department of Oral Pathology, School of Dentistry, University of Alabama at Birmingham 35294.
D018258:Herpesvirus 2, Human / Q000235:genetics; D004854:Herpesvirus 4, Human / Q000235:genetics; D006801:Humans; D009059:Mouth Diseases / Q000175:diagnosis* / Q000382:microbiology*; D009061:Mouth Mucosa / Q000382:microbiology; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics*; D030361:Papillomavirus Infections / Q000175:diagnosis*; D016133:Polymerase Chain Reaction; D018139:Simplexvirus / Q000235:genetics; D013283:Stomatitis, Herpetic / Q000175:diagnosis