Cervical Brush Research Articles

Anal Human Papillomavirus Testing with Digene's Hybrid Capture 2 Using Two Different Sampling Methods

The incidence of human papillomavirus detection in the anal canal is rising. Efficient anal screening by cytology is hampered because of poor specificity. Human papillomavirus (HPV) testing is proposed in addition to Papanicolaou (Pap) testing for the detection of cervical neoplasia. The purpose of this study was to determine the usefulness of a human papillomavirus-DNA detection test to detect human papillomavirus-associated disease and to compare two different methods of sample collection. In 555 patients, anal samples were obtained by using a cervical brush and a Dacron swab to test for high-risk and low-risk human papillomavirus-DNA. Patients positive for human papillomavirus-DNA underwent anoscopy. Biopsies were taken from visible lesions. Low-risk human papillomavirus-DNA was found in 325 of 555 patients (58.6 percent) and high-risk human papillomavirus-DNA in 285 of 555 patients (51.4 percent). Positive results confined to one single test method were higher for Dacron swab sampling (2.3 vs. 4.3 percent for low-risk human papillomavirus, P < 0.0001; 3.1 vs. 4.9 percent for high-risk human papillomavirus, P < 0.001). A positive correlation of relative light units was found for both sampling methods in the total cohort (P < 0.0001) as well as for patients who tested human papillomavirus-positive by both sampling techniques (P < 0.0001). Sampling with Dacron swabs yielded higher relative light units values compared with sampling with cervical brush for low-risk human papillomavirus-DNA and high-risk human papillomavirus-DNA. Anal screening for human papillomavirus-DNA by hybrid capture 2 is a useful method for detection of human papillomavirus-associated disease. Sample collection using Dacron swabs identifies more human papillomavirus-positive patients, and yields higher relative light unit values than using the cervical brush. Further studies are needed to determine the exact value of hybrid capture 2 in the screening for (pre)cancerous lesions of the anal canal.

Evaluation of DAPK gene methylation and HPV and EBV infection in cervical cells from patients with normal cytology and colposcopy

Evaluation of promoter methylation of the death-associated protein kinase (DAPK) gene and HPV and EBV infections in cervical cells from patients with normal cytology and colposcopy. Twenty women, who had been patients at the Institute of Gynecology of the Federal University of Rio de Janeiro (UFRJ) for routine examinations and who showed normal cytology and colposcopy, were selected for this work. Cervical brushings were used for DNA extraction, and the analysis of methylation patterns of the DAPK gene was done through chemical modification with sodium bisulfite. Analysis of viral infection was done using polymerase chain reaction (PCR). Of the 20 patients studied, six (30%) presented methylation of the DAPK gene, five (25%) presented infection with EBV and three (15%) presented coinfection with HPV/EBV. Associating methylation with viral infection, we found methylated DAPK in one patient (16%) with EBV, in two patients (33%) with co-infection and in three patients (50%) with no viral infection. In the present study, we verified, for the first time, the methylation pattern of the DAPK gene in cervical smears from patients with normal cytology and colposcopy. The results also showed the presence of viral infections in these patients. EBV infection, irrespective of whether associated with HPV or not, may contribute to cervical carcinogenesis as a cofactor. Methylation of the DAPK gene is associated with cell transformation, suggesting that DAPK methylation might be an important marker for the development of cervical epithelial neoplasias.

Detection of High-Risk Papillomavirus DNA with Commercial Invader-Technology-Based Analyte-Specific Reagents following Automated Extraction of DNA from Cervical Brushings in ThinPrep Media

We compared the performances of the Third Wave Technology Invader method and the Digene Hybrid Capture 2 assay to detect high-risk human papillomaviruses in 87 cervical brushing specimens submitted in Cytyc ThinPrep media. Two different methods for the extraction of DNA from squamous epithelial cells were also evaluated.

Potential Applications of Oral Brush Cytology with Liquid-based Technology: Results from a Cohort of Normal Oral Mucosa

Fifty healthy volunteers were studied to assess the potential applications of oral brush sampling using liquid-based cytology. Three specimens from the buccal mucosa and lateral border of tongue were collected from each subject by using cervical brushes and brooms. The brush was immersed in a preservative fluid. The sample in the preservative fluid was processed according to the manufacturer's directions (SurePath, UK). Slides were stained by the Papanicolaou method and assessed for squamous cell adequacy by the same criteria used for cervical cytology screening. Immunocytochemical staining for FHIT (Fragile Histidine Triad) was applied in liquid-based preparations following the streptavidin-biotin-peroxidase method. Human papillomavirus (HPV) detection was performed using the Hybrid Capture 2 assay (Digene) and the PCR-based Roche AMPLICOR HPV Test. LBC preparation slides showed good sample preservation, specimen adequacy and visualization of cell morphology. Interestingly, nine cases showed borderline cytological abnormalities from apparently normal oral mucosa. All cases showed good quality positive FHIT immunoreactivity staining. All studied cases were high-risk HPV negative using HC2 assay method. However, the AMPLICOR Roche Test detected four samples with positive results for high-risk HPVs. Liquid-based cytology has potential as a screening tool for oral cancer and precancer. The method may also have applications for research and practice in the field of oral cancer and precancer. However a special custom-designed oral cytobrush is required.

Assessment of MagNA Pure LC Extraction System for Detection of Human Papillomavirus (HPV) DNA in PreservCyt Samples by the Roche AMPLICOR and LINEAR ARRAY HPV Tests

Roche Molecular Systems recently released two PCR-based assays, AMPLICOR and LINEAR ARRAY (LA), for the detection and genotyping, respectively, of human papillomaviruses (HPVs). The manual specimen processing method recommended for use with both assays, AmpliLute, can be time-consuming and labor-intensive and is open to potential specimen cross-contamination. We evaluated the Roche MagNA Pure LC (MP) as an alternative for specimen processing prior to use with either assay. DNA was extracted from cervical brushings, collected in PreservCyt media, by AmpliLute and MP using DNA-I and Total Nucleic Acid (TNA) kits, from 150 patients with histologically confirmed cervical abnormalities. DNA was amplified and detected by AMPLICOR and the LA HPV test. Concordances of 96.5% (139 of 144) (kappa=0.93) and 95.1% (135 of 142) (kappa=0.90) were generated by AMPLICOR when we compared DNA extracts from AmpliLute to MP DNA-I and TNA, respectively. The HPV genotype profiles were identical in 78.7 and 74.7% of samples between AmpliLute and DNA-I or TNA, respectively. To improve LA concordance, all 150 specimens were extracted by MP DNA-I protocol after the centrifugation of 1-ml PreservCyt samples. This modified approach improved HPV genotype concordance levels between AmpliLute and MP DNA-I to 88.0% (P=0.043) without affecting AMPLICOR sensitivity. Laboratories that have an automated MP extraction system would find this procedure more feasible and easier to handle than the recommended manual extraction method and could substitute such extractions for AMPLICOR and LA HPV tests once internally validated.

Covariates of Cervical Cytokine mRNA Expression by Real-Time PCR in Adolescents and Young Women: Effects of Chlamydia trachomatis Infection, Hormonal Contraception, and Smoking

Measurements of mucosal immune parameters in the uterine cervix are potentially influenced by numerous factors, including infections, endogenous and exogenous hormones, semen, and nicotine and its metabolites in cervical mucus. The objective of this study was to examine correlates of immunoregulatory cytokine mRNA expression in cervical cytology samples in a cross-sectional design. Samples, collected at study entry by cervical cytology brush from 368 women aged 13-21 enrolled in a longitudinal study of the natural history of human papillomavirus (HPV) infection, were tested by quantitative RT-PCR for expression of IFN-gamma, IL-4, IL-10, and IL-12. In a multivariate analysis, elevated levels of IFN-gamma, IL-10, and IL-12 were significantly (p < 0.05) associated with several variables, including current C. trachomatis infection, recent intercourse, and current oral contraceptive pill use. Suppressed IL-4 and IL-10 levels were associated with cigarette smoking within the last 24 h. Time since last menstrual period did not affect any of the cytokines; in a substudy of weekly cytokine variability, however, IL-10 showed a non-significant trend toward higher levels around the time of menstruation.

Potential applications of oral brush cytology with liquid-based technology: Results from a cohort of normal oral mucosa

Fifty healthy volunteers were studied to assess the potential applications of oral brush sampling using liquid-based cytology. Three specimens from the buccal mucosa and lateral border of tongue were collected from each subject by using cervical brushes and brooms. The brush was immersed in a preservative fluid. The sample in the preservative fluid was processed according to the manufacturer's directions (SurePath, UK). Slides were stained by the Papanicolaou method and assessed for squamous cell adequacy by the same criteria used for cervical cytology screening. Immunocytochemical staining for FHIT (Fragile Histidine Triad) was applied in liquid-based preparations following the streptavidin-biotin-peroxidase method. Human papillomavirus (HPV) detection was performed using the Hybrid Capture 2 assay (Digene) and the PCR-based Roche AMPLICOR HPV Test. LBC preparation slides showed good sample preservation, specimen adequacy and visualization of cell morphology. Interestingly, nine cases showed borderline cytological abnormalities from apparently normal oral mucosa. All cases showed good quality positive FHIT immunoreactivity staining. All studied cases were high-risk HPV negative using HC2 assay method. However, the AMPLICOR Roche Test detected four samples with positive results for high-risk HPVs. Liquid-based cytology has potential as a screening tool for oral cancer and precancer. The method may also have applications for research and practice in the field of oral cancer and precancer. However a special custom-designed oral cytobrush is required.

Estudio comparativo de detección del virus papiloma humano (VPH) en muestras citológicas y biopsias de cuello uterino

The DNA quality for the detection and typification of Human Papilloma Virus (HPV) varies according to the type of sample in which it is studied. This may affect the sensitivity and specificity of the method employed. To study the yield and specificity of HPV detection and typification in uterine cervical samples obtained by cervical brushing fresh frozen and formalin fixed tissue. Cytological, fresh frozen and fixed tissue samples from 44 patients (nine with low grade lesions and 35 with high grade lesions) were studied. Nested polymerase chain reaction for genes E6/E7 was used to typify HPV groups as low risk or high risk. Of all the cytological samples obtained by brushing 84% of fixed samples and 43% of fresh frozen samples were positive for HPV. The yields were significantly different when comparing brushing with fixed tissue or fresh frozen tissue and fixed tissue with fresh frozen tissue (p <0.05). The frequency of high risk HPV fluctuated from 41% in fresh frozen tissue to 98% in cytological samples. Low risk HPV was detected in 16% of fresh frozen tissue and 68% of cytological samples. A mixed infection was detected in 66%, 41% and 14% of cytological, fresh frozen and fixed tissue samples respectively. Cytological samples obtained by brushing had the highest yield for the detection of cervical infection with HPV.

Determination of Human Papillomavirus (HPV) Load and Type in High-Grade Cervical Lesions Surgically Resected from HIV-Infected Women during Follow-up of HPV Infection

The role of human papillomavirus (HPV) load and the importance of multiple-strain HPV infections as biomarkers for the development of cervical disease were evaluated in human immunodeficiency virus (HIV)-positive women. A total of 108 samples were analyzed, 64 of which were obtained from 16 HIV-positive women who underwent surgical resection of the cervical cone for treatment of a histologically confirmed high-grade cervical intraepithelial neoplasm (cases) and 44 of which were obtained from 22 HIV-positive women who had high-risk HPV but a negative colposcopy result (controls). Each patient underwent periodic examinations at 6-12-month intervals that included colposcopy, Papanicolaou testing, biopsy (if indicated), and cervical brushing for HPV testing. Viral typing was performed by reverse dot-blot hybridization and quantification of viral load by in-house real-time PCR and commercial assays. Analysis of the cervical-brush samples collected when high-grade squamous intraepithelial lesions were diagnosed revealed that all cases had HPV loads that were significantly higher than those of controls (P=.0004 and P=.0003, by PCR and the Hybrid Capture 2 index [Digene], respectively). Decreasing concentrations of HPV load were observed when comparing samples obtained before and after treatment (P<.0001). The number and type of HPV strains that were detected were not statistically different between cases and controls. The significantly higher HPV load detected in women with high-grade cervical dysplasia, as well as the dramatic decrease in the load after surgical removal of the lesion, suggest that HPV load is a possible prognostic marker of high-grade SIL.

Impact of Chlamydia trachomatis and HPV Infection Among Sexually Active Teenage Girls in Upper Silesia, Poland

In their paper published in J Korean Med Sci entitled Screening for Chlamydia and gonorrhea by strand displacement amplification in homeless adolescents attending youth shelters in Korea, Lee et al. demonstrated that the prevalence of Chlamydia trachomatis among homeless adolescents was 12.6% (1). They reported a higher rate (28.1%) of infection among sexually active adolescents than in a sexually inactive group. In our study, to recognize the global importance of sexually transmitted infections (STI) among teenage girls in Upper Silesia we investigated 48 non-pregnant, sexually active girls aged 16-19, living in an urban area, who paid a visit for periodical check-up during oral contraceptive use. The majority of the adolescents reported periodical mild pain in the lower abdominal area and an increased amount of mucous discharge. They also reported early first sexual unprotected intercourse (<16 yr of age) and more than three sexual partners. We used direct immunofluorescence assay for C. trachomatis antigen detection (Chlamydia Direct IF, bioMerieux, France) and obtained a high prevalence (27%) of C. trachomatis infection. C. trachomatis infection is the most common bacterial STI, however, up to 70% of such infections are asymptomatic (2). The detection of asymptomatic infection has an important epidemiological implication, because the main consequences (infertility, ectopic pregnancy, pelvic inflammatory disease, and perinatal infections) of symptomatic and asymptomatic infections are often identical. In regions where an early screening program for detection of C. trachomatis infection has been established, the prevalence of infection in women is low: among Norwegian girls aged between 16-19 yr C. trachomatis infection was confirmed only in 9% (3). Testing for C. trachomatis in teenage girls is not routinely performed in many countries (including Poland). Cook et al. (4) found that only one-third of physicians carries out C. trachomatis tests in asymptomatic, sexually active teenage women during a routine gynecologic examination. In our previous study (5) of 205 sexually active women aged 17-40, we demonstrated a significant decrease of C. trachomatis infection among women younger than 25. British authors described 12.1% chlamydial infections in the age group of girls aged 16-19, 8.8% in the age group 20-24, and only 7.5% in the group younger than 16 (6). It is well known that human papilloma virus (HPV) is a proliferative virus, replicating only in actively dividing cells. The coexistence of bacterial infection (including C. trachomatis) may potentiate the proliferation and replication (7, 8). Moreover, C. trachomatis infection may be a possible cofactor of HPV infection, playing a major role in the etiology of cervical intraepithelial neoplasia. For this reason in our study group of teenage girls we also detected HPV DNA in collected cervical brushes using the Hybrid Capture I HPV Assay (Digene Corp. Beltsville, MD, U.S.A.), with complimentary probes for low oncogenic (6/11/42/43/44) and high oncogenic types (16/18/31/33/35/45/51/52/56). The collected cervical swabs were also assessed using the 2001 Bethesda System. HPV DNA was detected only in 2 cases with a normal cytology. Low grade squamous intraepithelial lesions (LSIL) was described in 5 cases out of 48, all of which were HPV DNA-negative, however 4 were C. trachomatis-positive. The role of C. trachomatis in the development of cervical dysplasia is still controversial. Antilla et al. (9), and Koskela et al. (10) considered an independent role of C. trachomatis in pathogenesis of cervical intraepithelial dysplasia. Our results, obtained on a small group of girls in Upper Silesia, were concordant with the above conclusion, however, further studies on larger group of adolescent girls are required to confirm this hypothesis.

Genetic damage in exfoliated cells from oral mucosa of individuals exposed to X-rays during panoramic dental radiographies

The genotoxic effects of X-ray emitted during dental panoramic radiography were evaluated in exfoliated cells from oral epithelium through a differentiated protocol of the micronucleus test. Thirty-one healthy individuals agreed to participate in this study and were submitted to this procedure for diagnosis purpose after being requested by the dentist. All of them answered a questionnaire before the examination. Cells were obtained from both sides of the cheek by gentle scrapping with a cervical brush, immediately before the exposure and after 10 days. Cytological preparations were stained according to Feulgen–Rossenbeck reaction and analyzed under light and laser scanning confocal microscopies. Micronuclei, nuclear projections (buds and broken eggs) and degenerative nuclear alterations (condensed chromatin, karyolysis and karyorrhexis) were scored. The frequencies of micronuclei, karyolysis and pycnosis were similar before and after exposure ( P > 0.90), whereas the condensation of the chromatin and the karyorrhexis increased significantly after exposure ( P < 0.0001). In contrast, both bud and broken egg frequencies were significantly higher before the examination ( P < 0.005), suggesting that these structures are associated to the normal epithelium differentiation. The results suggest that the X-ray exposure during panoramic dental radiography induces a cytotoxic effect by increasing apoptosis. We also believe that the score of other nuclear alterations in addition to the micronucleus improves the sensitivity of genotoxic effects detection.

A population-based study of cervical carcinoma and HPV infection in Latvia

Objectives. We wished to quantify the population-based importance of cervical carcinoma risk factors in Latvia. Methods. Totally, 223 of 224 eligible cases of incident invasive cervical carcinoma were enrolled during July 1998–February 2001 in Latvia. An age-matched sample of 300 healthy control women was selected from the Latvian population registry and 239 of these women (79%) were enrolled. A demographic and life-style questionnaire was completed, cervical brush samples were analyzed for human papillomavirus (HPV) DNA by PCR and serum samples for HPV antibodies. Results. Risk factors for cervical cancer in multivariate analysis were HPV type 16 or 18 DNA positivity (OR = 32.4; CI 95% 16.5–63.6) and living in the capital (OR = 2.4; CI 95% 1.2–4.7). Oral contraceptive use was not a risk factor (OR = 0.4; CI 95% 0.2–1.1). A strong protective effect was found for having had more than three Pap smears in the last 5 years (OR = 0.07 CI 95% 0.03–0.19). Conclusions. Inadequate population coverage of Pap smears, in spite of excessive smear usage, caused 28.4% of cervical cancers in age groups eligible for screening. HPV type 16 infection was the most important risk factor for cervical cancer in Latvia, with a population-attributable risk percent for all ages of 58.5%.

Depressed type 1 cytokine synthesis by superantigen-activated CD4+ T cells of women with human papillomavirus-related high-grade squamous intraepithelial lesions.

Carcinoma of the cervix is causally related to infection with the human papillomavirus (HPV), and T cells play a pivotal role in the immune response of the host to rid itself of HPV infection. Therefore, we assessed the T-cell function of women with HPV-related cervical neoplasia against a superantigen, Staphylococcus enterotoxin B (SEB). Each woman provided a cervical brush specimen for HPV DNA testing and Papanicolaou (Pap) smears for the staging of cervical lesions. They also provided a blood specimen for determination of the ability of CD4(+) T and CD8(+) T cells to synthesize Th1 (interleukin-2 [IL-2], gamma interferon [IFN-gamma], and tumor necrosis factor alpha [TNF-alpha]) and Th2 (IL-10) cytokines in response to activation with SEB. Compared with control subjects with self-attested negative Pap smears, women with high-grade squamous intraepithelial lesions (HSIL) had significantly lower percentages of activated CD4(+) T cells that produced IL-2 (P = 0.045), IFN-gamma (P = 0.040), and TNF-alpha (P = 0.015) and a significantly lower percentage of activated CD8(+) T cells that produced IL-2 (P < 0.01). These data indicate that women with HPV-related cervical HSIL show a decrease in Th1 cytokine production by activated CD4(+) T cells and suggested that compromised T-helper functions may negatively impact the function of cytotoxic CD8(+) T cells.

Comparación del cepillo endocervical y la torunda de algodón para la toma citológica cervical

ObjectiveTo compare the quality and diagnostic yield of cervical cytologic sampling according to the Bethesda System 1988–1991 based on the use of an endocervical brush or cotton swab. Material and methodsWe performed a randomized study of 595 women distributed in two groups. Cervical sampling was performed with a cotton swab and wooden spatula in group A (305 women) and with a cervical brush and wooden spatula in group B (290 women). ResultsThere were no statistically significant differences in the clinical or demographic features of the two groups. No differences were found in the effectiveness of the two sampling methods in detecting cytological alterations or collection of endocervical cells. Satisfactory smears for interpretation were obtained using both methods. ConclusionNo change in the traditional method of cervical sampling is required. Attempts to improve the quality and false-negative rate of samples should focus on other features.

Acetic-acid guided visual inspection vs. cytology-based screening for cervical cancer in the Philippines

Objectives: To compare the validity and acceptability of acetic-acid visualization (VIA), magnified acetic-acid visualization (VIAM), spatula+cotton swab–Papanicolaou (Pap) smear (SS), and cervical brush–Pap smear (CB) in the detection of precursor/early cervical cancer lesions. Methods: A total of 12 992 women aged between 25 and 65 years from 14 Philippine centers were randomly allocated to the four tests. The gold standard was colposcopy with biopsy for positive/suspicious cases. Results: Sensitivity rates [95% confidence intervals (CIs)] were 37 (CI, 26.8–48.5), 34.1 (CI, 24.8–44.8), 14.3 (CI, 6.4–27.8), and 19.1 (CI, 9.2–34.6) for VIA, VIAM, SS, and CB, respectively. Specificity rates were 90.7 (CI, 89.6–91.7), 90.7 (CI, 89–91.1), 97.5 (CI, 96.8–98), and 97.9 (CI, 97.3–98.4), respectively. Kappa for the Pap smear (PS) within centers ranged from −0.154 to 0.783, and between centers from −0.028 to 0.364. Screeners preferred CB; screened-women preferred VIA. Conclusions: The acetic-acid visualization and VIAM methods are recommended for initial cervical cancer screening in the Philippines.

Optimizing the hybrid capture II human papillomavirus test to detect cervical intraepithelial neoplasia

OBJECTIVE: To estimate the optimal relative light unit ratio, and correspondingly viral load, of the hybrid capture II oncogenic human papillomavirus deoxyribonucleic acid test for detecting cervical intraepithelial neoplasia (CIN). METHODS: Women with abnormal cytology were referred for colposcopy, and a cervical swab or brush specimen was obtained for human papillomavirus testing. Sensitivities, specificities, and likelihood ratios of different relative light unit ratio cutoffs were calculated using a reference standard of colposcopy or biopsy of either CIN II+ (CIN II, III, or carcinoma), or CIN I+ (CIN I, CIN II+). The receiver operating characteristic curve was used to estimate optimal test-positive cutoff points for the hybrid capture II test. RESULTS: CIN II+ was found in 18.7% (98 of 524) and CIN I in 10.5% (55 of 524) of the women. The optimal relative light unit ratio was 15.56, giving a sensitivity and specificity of 82.7% and 73.2% for CIN II+, and 74.2% and 77.8% for CIN I+. In a stratified analysis, a higher relative light unit cutoff (15.19) optimized sensitivity and specificity for CIN II+ (sensitivity 81.8%, specificity 51.5%) for women with low-grade squamous intraepithelial lesions cytology, whereas the optimal cutoff was 2.36 (sensitivity 100%, specificity 73.0%) for women with atypical squamous cells of undetermined significance, yielding referral rates of 53.3% and 28.7%, respectively. Use of a swab or brush was not associated with the level of human papillomavirus detected ( P > .05). CONCLUSION: Use of a higher cutoff for the relative light unit ratio (higher viral load) of the hybrid capture II test may improve the management of women, especially those with low-grade squamous intraepithelial lesions cytology.

Detection of human herpes virus type 6 DNA in precancerous lesions of the uterine cervix.

Human herpes virus type 6 (HHV-6) DNA has been suggested to be a cofactor to human papillomavirus (HPV) in cervical cancer. In a cross-sectional study, we investigated the association between HHV-6 DNA detected in cervical brushings and high-grade squamous intraepithelial lesions (HSIL), while controlling for genital infection with 27 genotypes of HPV. Of the 320 women recruited from an oncologic gynecology clinic, 50 had invasive cervical cancer, 65 had HSIL, 80 had low-grade squamous intraepithelial lesions (LSIL), and 125 were normal. Four of the seven HHV-6-positive women had HSIL. HHV-6 was associated with HSIL after adjusting for age and socioeconomic status (odds ratio [OR] of 10.9, 95% confidence interval [CI]: 1.1-107.1). This association was no longer significant after controlling for HPV (OR = 6.4, 95% CI = 0.3-128.5). HHV-6 was detected in cervical samples from women with precancerous and cancerous lesions of the cervix, but not significantly more frequently than in normal women.

Human papillomavirus testing and the management of women with mildly abnormal cervical smears: an observational study

Editorial by Manos Following publication of a report in Health Technology Assessment the NHS is running a pilot scheme screening women for human papillomavirus if they have a mildly dyskaryotic...

Comparison of cervical smears secured with different instruments

In this presentation we have compared the results from cytologic samples obtained with the cervical brush and Ayre's wooden spatula, against samples obtained with Acellon Combi. One thousand cervical smears were collected. This was a cross-sectional study, where it was decided randomly to use cervical brush for sampling of the endocervix and Ayre's wooden spatula for exocervix or Acellon Combi for both. The combination of the cervical brush and the wooden spatula yielded an acceptable quantity of columnar cells in 71.6% of the cases, during both endocervical and exocervical sampling, while the Acellon Combi yielded an acceptable quantity in 61.7% of the cases (p<0.01). The combination of brush/spatula yielded an acceptable quantity of squamous cells from samples of endocervix and exocervix in 87.5% cases, while with the Acellon device amounted to 77.4% (p<0.001). Highly significant differences in the state of cellular preservation of samples from endocervix obtained with cervical brush technique were found upon comparison to Acellon group. Henceforth, we believe that the use of the cervical brush and the Ayre wooden spatula increases the quantity and preservation of the cells retrieved during cervical smears compared to Acellon Combi.

Spectral imaging of multi-color chromogenic dyes in pathological specimens.

We have investigated the use of spectral imaging for multi‐color analysis of permanent cytochemical dyes and enzyme precipitates on cytopathological specimens. Spectral imaging is based on Fourier‐transform spectroscopy and digital imaging. A pixel‐by‐pixel spectrum‐based color classification is presented of single‐, double‐, and triple‐color in situ hybridization for centromeric probes in T24 bladder cancer cells, and immunocytochemical staining of nuclear antigens Ki‐67 and TP53 in paraffin‐embedded cervical brush material (AgarCyto). The results demonstrate that spectral imaging unambiguously identifies three chromogenic dyes in a single bright‐field microscopic specimen. Serial microscopic fields from the same specimen can be analyzed using a spectral reference library. We conclude that spectral imaging of multi‐color chromogenic dyes is a reliable and robust method for pixel color recognition and classification. Our data further indicate that the use of spectral imaging (a) may increase the number of parameters studied simultaneously in pathological diagnosis, (b) may provide quantitative data (such as positive labeling indices) more accurately, and (c) may solve segmentation problems currently faced in automated screening of cell‐ and tissue specimens. Figures on http://www.esacp.org/acp/2001/22‐3/macville.htm.