Search

Self-collected vaginal specimens tested for high-risk human papillomavirus (HR-HPV) have been shown to be less sensitive for the detection of cervical intraepithelial neoplasia or cancer (≥CIN 3) than physician-collected endocervical specimens. To increase the sensitivity of self-collected specimens, we studied a self-sampling device designed to obtain a larger specimen from the upper vagina (POI/NIH self-sampler) and a more sensitive polymerase chain reaction (PCR)-based HR-HPV assay. Women (10,000) were screened with cervical cytology and HR-HPV testing of vaginal self-collected and endocervical physician-collected specimens. Women were randomly assigned to use either a novel self-collection device (POI/NIH self-sampler) or conical-shaped brush (Qiagen). The self-collected and clinician-collected specimens were assayed by Cervista (Hologic) and the research only PCR-based matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF). Women with any abnormal screening test underwent colposcopy and biopsy. Women (8,556), mean age of 38.9, had complete data; 1.6% had ≥ CIN 3. For either HR-HPV assay, the sensitivity was similar for the two self-collection devices. Tested with Cervista, the sensitivity for ≥CIN 3 of self-collected specimens was 70.9% and for endocervical specimens was 95.0% (p = 0.0001). Tested with MALDI-TOF, the sensitivity for ≥CIN 3 of self-collected specimens was 94.3% and for endocervical specimens was also 94.3% (p = 1.0). A self-collected sample using a PCR-based assay with the capability of very high throughput has similar sensitivity as a direct endocervical specimen obtained by a physician. Large population-based screening "events" in low-resource settings could be achieved by promoting self-collection and centralized high-throughput, low-cost testing by PCR-based MALDI-TOF., (Copyright © 2011 UICC.)

The aim of this study was to evaluate the accuracy of the ThinPrep Imaging System (Imager) in detecting atypical glandular cells (AGC) or adenocarcinoma in the 22 selected fields. All cases reported as AGC or adenocarcinoma from January 2005 to December 2006 that had been initially screened by the Imager were retrospectively reviewed to determine whether the most diagnostically relevant groups/cells were within the 22 selected fields. A total of 39 cases were reviewed. The cases were divided into two groups: the group with diagnostic cells detected within the 22 selected fields (accurately detected group) and the group with upgraded diagnosis following rescreening process (underdetected group). The Imager accurately detected 32 (82%) of cases with abnormal glandular cells, including six cases reported as adenocarcinoma, one case as adenocarcinoma in situ (AIS), and 25 cases as AGC. In seven (18%) cases, the Imager failed to detect the most abnormal cells within the 22 selected fields. Among these, one case was adenocarcinoma, while the rest were reported as AGC. Overall, four cases were assessed as atypical during quality control (QC) rescreening even though the Imager detected abnormal groups in most. Fourteen of 32 cases had abnormalities proven by histologic follow-up. Overall, the Imager was effective in detecting most AGC, AIS and invasive adenocarcinomas. In a minority (18%) of cases, the Imager failed to detect the cells of interest within the 22 selected fields. However, full manual review of cases with potential atypical/reactive groups or endometrial groups in women > or = 40 and QC rescreening of selected cases may help to minimize the underdetected cases., (2009 Wiley-Liss, Inc.)

The Cytyc ThinPrep Imaging System was FDA approved based on a multi-institutional clinical trial, in which the HSIL+ prevalence rate was 0.7%. This study determines the effectiveness of the Imager in clinical practice at an academic medical center with a historical HSIL+ rate of 0.25%. Cytological interpretations were compared for two 12-month periods pre- and post-Imager implementation. Data was compiled by cytologic diagnoses, and variations in prevalence rates were analyzed for statistical significance. Interpretations of ASC-US, ASC-H, and LSIL were correlated with Digene Hybrid Capture2 High Risk HPV DNA testing; interpretations of ASC-H, LSIL, and HSIL+ were correlated with subsequent surgical follow-up. ASC-US, ASC-H, and LSIL detection rates increased 34, 48, and 29%, respectively, with the Imager (P<0.001); whereas the detection of HSIL increased 24% (P<0.051). Surgical correlation revealed no statistical differences in the positive predictive value (PPV) for ASC-H and LSIL. However, an increase in the PPV of HSIL was found (P<0.05). High risk HPV results were lower for ASC-US (P<0.001), but statistically equivalent for ASC-H and LSIL. Results of surgical correlation and HPV testing validated an increase in detection rates of ASC-H, LSIL, and HSIL, as well as increased PPV of HSIL with the ThinPrep Imaging System.

Despite the fact that compact instruments dedicated to counting cells are readily available especially in hematology, liquid-based cervical cytology specimen adequacy is still determined by pathologist review following the preparation of slides. More than 100 liquid-based cervical cytology specimens collected in ThinPrep vials were processed into slides for diagnosis and assessment of specimen adequacy using the Bethesda 2001 criteria. Residual liquid (100 muL) was transferred to a single well of a 96-well plate and run in a capillary cytometer (Guava Technologies, Hayward, CA) with forward scatter and side scatter detectors. We determined that the sensitivity and specificity of this assay for unsatisfactory samples was 100% and 97%, respectively, compared with cytopathologic examination. The difference in the concentration of ectocervical cells/muL (EPU) between samples determined to be unsatisfactory by virtue of less than an estimated 5,000 cells per slide and satisfactory samples were statistically significant (P< 0.001). Here, we report the use of this rapid specimen adequacy assay on a small, capillary based, personal cytometer. Use of this instrument reduced the amount of sample required by >90% and reduced the average per sample assay time from 120 s to 10 s compared with flow cytometry., ((c) 2007 Clinical Cytometry Society)

The aim of this study was to evaluate the efficacy of endometrial cytology using the Uterobrush in the collection of samples for the diagnosis of endometrial lesions. The preparation technique for endometrial brushing specimens was also demonstrated. In their earlier study, the authors described the cyto-architectural criteria that appear to be more useful for the cytological assessment of endometrial lesions. Therefore, for the application of the diagnostic criteria, endometrial cytological sampling will become more important. With regard to the cell sample collection, the authors used the Uterobrush because the insertion into the uterine cavity is easy and painless. The authors compared the Uterobrush with the Endocyte, since they thought that cell clumps using the former device tended to be smaller. The purpose of the current study was to improve and evaluate cell preparation methods using the Uterobrush. The authors investigated three methods [i.e., conventional and improved techniques ("flicked" method) with the Uterobrush and the Endocyte] of endometrial cell collection and preparation. Using conventional methods, a brush was rolled along a glass slide and the collected material spread and smeared. However, using the "flicked" method, a brush is strongly flicked with forceps, so that the cells are transferred to the slide and its position is moved along the slide little by little and smeared. The frequency by size of cell clumps with tube or sheet-shaped pattern was examined in the preparations. Cell block specimens with the Uterobrush were also investigated. Endometrial cytology from a total of 90 women was evaluated. Most were outpatients and all were older than 20 yr (ranging from 20 to 54, average 42.7 yr). Of these, 30 cases from each group were examined by three methods. Uterobrush samples prepared by the "flicked" method have a greater quantity of cell clumps than those using the Endocyte sampler, while the frequency-by-size of cell clumps was by degree the same as the Endocyte. The cell clumps obtained in the Uterobrush "flicked" method preparation was considered equivalent or superior as an aid to making a diagnosis of endometrial lesions and it became obvious that the same criteria were applicable to both of instruments. Our cytological examination may be a potent aid to making a diagnosis of endometrial lesions and these findings will be helpful in the standardization of criteria in direct intrauterine cell samples., (Copyright 2006 Wiley-Liss, Inc.)

ThinPrep (TP) Papanicolaou (Pap) samples containing excessive blood often result in unsatisfactory preparations, possibly leading to undetected gynecologic disease, and added inconvenience to patients and clinicians. Reprocessing of these samples with a glacial acetic acid wash is effective at eliminating blood, providing satisfactory preparation and detection of lesions. However, it increases laboratory costs and decreases work flow efficiency. We report the use of a color standard for gauging the necessity of performing a glacial acetic acid wash before TP processing. This "preprocessing" was found to reduce the costs associated with reprocessing by 48%, while maintaining high preparation quality by improved sample adequacy.

Background: Acquisition of DNA ploidy histograms by image analysis may yield important information regarding the behavior of premalignant cervical lesions. Accurate selection of nuclei for DNA measurement is an important prerequisite for obtaining reliable data. Traditionally, manual selection of nuclei of diagnostic and reference cells is performed by an experienced cytotechnologist. In the present study, a method for the fully automated identification of nuclei of diploid epithelial reference cells in Feulgen- restained Papanicolaou (PAP) smears is described., Methods: The automated procedure consists of a decision tree implemented on the measurement device, containing nodes with feature threshold values and multivariate discriminant functions. Nodes were constructed to recognize debris and inflammatory cells, as well as diploid and nondiploid epithelial cells of the uterine cervix. Evaluation of the classifier was performed by comparing resulting diploid integrated optical densities with those from manually selected reference cells., Results and Conclusion: On average, automatically acquired values deviated 2.4% from manually acquired values, indicating that the method described in this paper may be useful in cytometric practice., (Copyright 2002 Wiley-Liss, Inc.)

We report on the formulation and use of an alcoholic-agar additive solution generally useful for rapid, inexpensive liquid-based cytology slide preparation. Gynecological cytology specimens were collected from 1,000 women. Two hundred fifty aliquots each of CytoRich, CytoRich Red (Tripath Imaging, Inc., Burlington, NC), Preservcyt (Cytyc Corp., Boxborough, MA), and DINA*TRANS (DINA*CYT Corp., Portland, OR) fixatives were used for this study. Fixed cell suspensions from 1,000 women, seen consecutively by a general obstetrics and gynecology practice, were vortex-mixed and transferred into a premeasured amount of alcoholic-agar in test tubes. Test tubes were conventionally centrifuged, cells were trapped in a spontaneously formed agar-gel, and the supernatant solutions were decanted. Vortex-mixing cell buttons caused a rapid gel-to-sol transition, affording viscous cell suspensions that were applied to slides, smeared, and stained using Papanicolaou stains. Slides showed unclumped, monolayered, uniform, random cell-spreads. All of the fixatives afforded crisp presentation of normal and abnormal cells. There was an about 3-fold increase in HSIL+ (0.7-1.8%) and LSIL diagnoses (1.3-4.4%), and a 45% reduction in ASCUS diagnoses (3.3-1.8%), as compared to our cytology laboratory's previous year's patients' statistics with a concurrent 0.2% unsatisfactory rate, due solely to inadequate sampling. This manual method makes liquid-based cytology inexpensive and does not require specialized preparative devices., (Copyright 2001 Wiley-Liss, Inc.)

Automated-primary screening is frequently performed, but conventional smears have to be rescreened on coverslip edges because automated systems do not analyze cells near the coverslip edges because of focusing problems. This disadvantage can be overcome by obtaining conventional samples smeared in the center of the slide, e. g., by the Exact Touch (ET), a single sampler introduced by G. Saccomanno. PAPNET-assisted primary screening was performed on 263 samples collected by ET. The smears were classified as "negative" or as "review." The negative cases were rapidly rescreened but not near the coverslip edges. The review cases were fully reanalayzed by the optic microscope. Ten positive smears were identified among the review cases: 3 smears showed changes due to human papillomavirus, 3 low-grade squamous intraepithelial lesions, and 4 high-grade squamous intraepithelial lesions. The detection of abnormal cells was easy by means of the combined use of ET and PAPNET. The ET-PAPNET technique was very useful in analyzing conventional smears and in reducing the time-consuming process of detecting abnormal cells., (Copyright 2000 Wiley-Liss, Inc.)

Federally-mandated quality control (QC) in Papanicolaou (Pap) smear testing requires rescreening of 10% of negative smears, to include cases selected randomly as well as smears from patients that may have a higher risk for developing cervical cancer based on clinical information. FDA approval of NeoPath's AutoPap 300 QC system (NeoPath, Inc., Redmond, WA) allows practical QC rescreening of all negatives. We tested the ability of AutoPap to help increase identification of detection errors compared to random 10%/high-risk selection. From March 1-August 30, 1997, we utilized AutoPap/high-risk status to select cases for manual rescreen, and compared the rate of identification of primary screening errors to that for the preceding year using 10% random selection/high-risk status. Of 35,027 smears accessioned, 31,240 (89.1%) were screened as negative and 7,965 were selected for manual rescreen. Of these, 353 were determined to be abnormal. Most abnormals identified by this protocol were classified as atypical squamous or glandular cells of undetermined significance (ASCUS or AGUS). However, 59 low-grade squamous intraepithelial lesions (LSIL) and 13 high-grade squamous intraepithelial lesions (HSIL), many with few abnormal cells, were also identified. These results represented an increase in pickup rate of false negative due to detection errors of 2.3-, 2.8- and 5.6-fold for atypical squamous or glandular cells of undetermined significance, LSIL, and HSIL, respectively, when accounting for the volume differences over the time period measured. Our findings strongly support the conclusions drawn from clinical trials of the AutoPap that false negatives due to detection error can be significantly reduced when using AutoPap as part of a routine quality control program.

The Papanicolaou stained cervicovaginal smear (Pap smear) is a gynecologic cytologic screening tool that has dramatically reduced the incidence and mortality of cervical cancer. Recently, a number of problems with the Pap smear test, including severe cytotechnologist shortages, lack of internal quality controls, and high false negative rates have been emphasized by the scientific community and the general public. To address some of these problems, there has been increased development of a number of automated and semiautomated cytologic screening systems. A semiautomated screening system (PAPNET) was used to retrospectively evaluate 527 conventionally prepared Pap smears from 500 consecutive unselected patients. This system scanned the slides and displayed 128 of the most "abnormal" areas of each slide on a monitor. A screener reviewed these "abnormal" images in a blinded fashion and decided whether they represented variants of normal cytology, were inadequate for evaluation, or were abnormal and warranted manual review of the material. The screener's evaluations were then compared to the previously made diagnosis and discrepancies were reviewed. After review of the images from 500 patients, 343 (69%) were deemed to be normal cytology, 140 (28%) were abnormal and needed manual review, and 17 (3%) were inadequate. Of the 343 called normal using this system, 338 were previously called normal and five patients were previously diagnosed with either atypical squamous cells of undetermined significance (ASQUS) or low grade squamous intraepithelial lesions (LGSIL). Of the 140 thought to need review, 43 were previously diagnosed with squamous or glandular disease and 97 were previously diagnosed as normal. Slides from the apparent false negatives and false positives were again reviewed in a blinded fashion.(ABSTRACT TRUNCATED AT 250 WORDS)

The cytologic distinctions between artifactual atypia induced by endocervical brush collection, endocervical glandular repair, and endocervical glandular dysplasia or neoplasia are often difficult. This difficulty is amplified by the lack of accepted cytologic criteria for endocervical glandular dysplasia, and the fact that earlier studies defining the significant cytologic features of endocervical adenocarcinoma in situ and endocervical adenocarcinoma (endocervical neoplasia) utilized a swab for endocervical cell retrieval. With the almost universal adoption of the endocervical brush, it is important to redefine those cytologic characteristics that differentiate true endocervical glandular dysplasia or neoplasia from reactive or reparative processes and instrument artifact. From our data, it appears that the classically described cytologic discriminators between a negative smear and smears of endocervical dysplasia or neoplasia are basically unchanged with some minor modifications for endocervical brush-collected samples.

An 8-mo (August 1988 through April 1989) study was performed to evaluate the efficacy of the modified Ayre spatula/Zelsmyr Cytobrush versus the modified Ayre spatula/bulb aspirator in the collection of cells, both normal and abnormal, from the uterine cervix. The majority (78%) of the 192 patients, ranging in age from 18 to 82 yr, were referred to the gynecologic/oncologic clinic for follow-up of an atypical Papanicolaou smear or of a previously diagnosed and treated cervical, vulvar, or endometrial lesion. Of the 192 patients, 149 were premenopausal (15 of whom were pregnant), 12 were perimenopausal, and 31 were postmenopausal. Patients were randomized into one of two groups based on the collection of samples for their cervical smears: 1) modified Ayre spatula (exocervix) plus Zelsmyr Cytobrush (endocervix: 90 cases) and 2) modified Ayre spatula (exocervic) plus bulb aspirator (endocervix; 102 cases). Of the smears prepared with the spatula/Cytobrush (90 cases), only 2 (2.2%) lacked endocervical cells. Within this group, the smears of 5 of 5 pregnant patients (100%) and 13 of 13 postmenopausal patients (100%) contained endocervical cells. Cytologic abnormalities were present in 15 of these cases; koilocytosis in 4, cervical intraepithelial neoplasia (CIN)-I in 5, CIN-II in 4, and CIN-III in 2. Five of the 15 cytologically positive cases had histologic verification; the cytologic findings were predictive of the histologic findings in all. Of the smears prepared using the spatula/bulb aspirator (102 cases), 26 (25.5%) lacked endocervical cells. Within this group, the smears of only 4 of 10 pregnant patients (40%) and 14 of 18 postmenopausal patients (78%) contained endocervical cells.(ABSTRACT TRUNCATED AT 250 WORDS)

A study was undertaken to evaluate the efficacy of the Cytobrush compared with the cotton-tipped applicator for obtaining cells from the uterine cervix. Four consecutive Papanicolaou smears were obtained in 1,000 patients during a single clinic visit. In every patient, a total of four smears on two slides were submitted: one slide contained endocervical material obtained with a cotton-tipped applicator and with the Cytobrush and the other slide contained ectocervical material obtained with both a cotton-tipped applicator and the Cytobrush. In endocervical smears, the Cytobrush technique produced numerous groups and clusters of cervical columnar cells in contrast to the use of the cotton swab. This was particularly true in older patients and pregnant women. In ectocervical smears, the detection of the squamous component was also higher in the Cytobrush smear than in cotton swab smears. However, the cell yield was not influenced by the cell sampling sequence in either endocervical or ectocervical specimens. It is the conclusion of this study that the utility of the Cytobrush technique for collecting specimens for endocervical and ectocervical smears is greater and provides a higher yield of cells than the cotton swab technique. Wider use of this simple procedure is encouraged.

From 10(1) to 10(6) TCID50 (mean tissue culture infective doses) per ml HSV I and II can be quantitatively recorded from vaginal tampons. Tampons and sterile cotton swabs are equally sensitive in recovering HSV I and II. Direct cotton swab cultures of the cervix and cervicovaginal tampon cultures from the same patients recovered similar quantities of HSV, ranging from 1.5 to 6.5 log10 TCID50/ml in 5 patients.

A new means of disaggregating cytology specimens in suspension using an immersible rotor device is described. The new rotor is compared to an automated syringing apparatus using cervical samples. Similar results using both devices are obtained for both normal and abnormal specimens.