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One hundred paraffin-embedded cervical biopsy specimens were tested for the presence of human papilloma virus (HPV) by in situ hybridization (ISH), and by direct and indirect in situ PCR (IS-PCR) in order to evaluate the efficiency of the different in situ methods in detecting HPV infection. ISH was performed using either commercial DNA probes or a cocktail of 5'-digoxigenin labeled oligoprimers. The same were used for ISH during indirect IS-PCR. To enhance the sensitivity of ISH several polymers, i.e., polyvinyl alcohol (PVA), polyethylene glycol, and polyvinylpyrrolidone were added to the alkaline phosphatase nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl phosphate (NBT/BCIP) reaction. Furthermore, tyramide signal amplification (TSA) was tried for signal amplification. Those samples treated with PVA during the NBT/BCIP reaction did not show any signal amplification whereas those treated with TSA exhibited a dramatic increase in sensitivity with usually acceptable signal to noise ratios. Our results show that, regarding sensitivity, ISH with subsequent signal amplification by TSA can be used as an almost equivalent alternative to the more cumbersome IS-PCR on routinely processed tissue specimens. When considering reproducibility, it is superior to IS-PCR.

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Cases of adenoid cystic carcinomas of the salivary (n = 90) and lacrimal glands (n = 6) from the years 1965-1980 were evaluated retrospectively with regard to clinical, epidemiologic and histomorphologic parameters, and in 52 cases, nuclear DNA content was assessed using a single cell scanning cytophotometry procedure in order to determine prognostic factors. Clinical courses were poor with a high incidence of recurrences, hematogenous metastases and deaths from tumor. Histology was related to prognosis, glandular tumors showing a better prognosis than solid ones. Tumor size greater than 4 cm was correlated with an unfavorable clinical course in all cases. Cytophotometry yielded various types of histograms (7 diploid, 10 proliferative, 14 triploid, 19 atypical, 2 tetraploid). Significant correlations were found as to the time of survival, tumors with diploid histograms showing the longest intervals and those with atypical ones the shortest. Although the prognosis of adenoid cystic carcinoma remains poor, cytophotometry can offer additional prognostic information in the individual case.