Time trends in the prevalence of human papillomavirus infections in archival Papanicolaou smears: Analysis by cytology, DNA hybridization, and polymerase chain reaction

Abstract

A retrospective study was undertaken to determine the prevalence of human papillomavirus (HPV) infections in routine Papanicolaou (Pap) smears collected by general practitioners from Western Australian women in each of the years 1972, 1982, and 1987. HPV infection was detected by cytology, dot-blot hybridization, or polymerase chain reaction (PCR). It was found that the prevalence of HPV infection remained unchanged over the 15 year study period, was independent of age, and was associated with normal cytology at a rate far greater than previously recognized. Indeed, the prevalence of cervical intraepithelial neoplasia (CIN) lesions, as detected by cytology, was 3.0% in 1972 and 3.8% in 1982 and 1987. The prevalence of HPV infection, detected as koilocytosis or parakeratosis, was 6.5%, 6.8%, and 5.3% in smears collected in 1972, 1982, and 1987, respectively, from 1,800 women. In 237 cytologically normal smears reprocessed for HPV-DNA studies, the prevalence of HPV 16 was determined to be 15.6%, 11.2%, and 17.8% in 1972, 1982, and 1987, respectively, as determined by dot-blot hybridization. However, the PCR detected HPV 16 in an additional 55.5%, 62.9%, and 57.0% of cytologically normal and dot-blot negative smears. The prevalence of HPV 16 infection in cytologically normal smears was estimated to be 71.0%, 74.1%, and 74.8% in 1972, 1982, and 1987, respectively, by combining the HPV 16 dot-blot and PCR-positive results. The high prevalence of HPV 16 in cytologically normal Pap smears suggests that infection with HPV 16, as detected by PCR amplification, does not place women in a high-risk category for cervical cancer.(ABSTRACT TRUNCATED AT 250 WORDS)

Processing of long-stored archival cervical smears for human papillomavirus detection by the polymerase chain reaction

“…These methods vary from relatively mild treatment, using a simple boiling and/or freeze-thaw step (Shibata et al., 1988; van den up to a complete DNA isolation procedure. Also, for the processing of archival cervical smears different methods have been applied, including DNA extraction after proteinase K lysis (Jackson et al, 1989;Rakoczy et al, 1990;Gall et al, 1993) and the application of guanidinium isothiocyanate (GTC) lysis and silica beads nucleic acid extraction (Smits et al, 1992). Although the use of these extraction methods revealed a successful PCR, little is known about the reproducibility and sensitivity of the PCR, which might be influenced by differences in DNA yield and quality of the samples.…”

Detection and typing of human papillomaviruses present in fixed and stained archival cervical smears by a consensus polymerase chain reaction and direct sequence analysis allow the identification of a broad spectrum of human papillomavirus types

“…For epidemiological purposes, methods based on the polymerase chain reaction (PCR) (Saiki et al, 1988) have been designed for the detection and typing of HPV DNA in fresh or frozen cervical biopsies (Li et al, 1988 ;Manos et al, 1989), unprocessed cervical smears (van den Brule et al, 1990), paraffin-embedded tissue specimens (Shibata et al, 1988;Cornelissen et al, 1989) and archival cervical smears (Rakoczy et al,, 1990). The prevalence of HPV in cervical smears from women participating in a screening programme for cervical cancer (van den Brule et al, 1991) and specific groups of patients (Young et al, 1989;Manos et al,, 1990;van den Brule et al, 1990van den Brule et al, , 1991 has been reported.…”

Optimal Recovery of DNA for Polymerase Chain Reaction–Based Assays from Fine Needle Aspirates

“…This is well exemplified by superior detection of infectious agents by these techniques, as for Pneumocystis carinii from sputum 40 and Mycobacterium tuberculosis from lymph node aspirates. 4,11,18 Initial reports have been related predominantly to human papillomavirus infection in cervical smears, 6,10,30,37 followed by detection of HPV from aspirates of metastatic cervical tumors. 24,33 The number of published articles combining PCR and cytology has increased exponentially in the last few years.…”

“…However, it seems that old specimens can be successfully studied by PCR. 3,15,23,30 Large amounts of melanin can inhibit PCR. 29 Although specimens obtained from case 7 showed rich cellular material, the beta-actin gene could be amplified only from a cell block and Papanicolaou samples, which demonstrated a weak signal.…”

“…24,33 The number of published articles combining PCR and cytology has increased exponentially in the last few years. 14 Studies have included use of fresh cellular specimens 2,4,13,[16][17][18]24,28,[36][37][38]41 ; previously stained, alcohol-fixed and air-dried smears 1,3,11,12,15,23,30,34 ; and cell block preparations 8,39 for PCR analysis to detect various alterations. Few studies have emphasized the comparison of results obtained from different types of preparations.…”