Direct-to-vial comparison of a new liquid-based cytology system, liqui-PREP versus the conventional pap smear

The objective of the study was to assess the clinical utility of a liquid-based cytology system on cervicovaginal screening in a clinical commercial laboratory. Twenty-six thousand, one hundred and seventy eight cervicovaginal specimens were prepared by the Liqui-PREP (LGM International Inc., Ft. Lauderdale, FL) with a direct-to-vial comparison to 218,548 cases of the conventional direct smear from August 2005 through December 2005. Biopsy data was obtained to confirm the sensitivity for each method. Liqui-PREP showed a marked increase in HSIL+ detection compared to the conventional smear ( P = 0.001). The rate of LSIL and AGC detection was higher with Liqui-PREP ( P equals; 0.001 for both). The percentage of ASCUS specimens was higher than with conventional smear due to cleaner slides and easier detection of suspicious cells. The WNL rate was lower for Liqui- PREP ( P = 0.001) consistent with increased HSIL+ and ASCUS. The unsatisfactory rate was lower for Liqui-PREP ( P = 0.017). The histological predictive value of Liqui-PREP was slightly higher than the conventional smear (94.1% versus 89.9%). The Liqui-PREP system similar to other reported LBC technologies shows an increased detection of squamous intraepithelial lesions, and gives higher-quality slides for interpretation than the conventional smear. Histological results confirm that this increase in cytological findings are clinically significant. LGM's new LBC technology is a more sensitive screening tool when compared with the conventional smear.     

Cervical biopsy-based comparison of a new liquid-based thin-layer preparation with conventional Pap smears

The objective of this study is to compare the diagnostic efficacy of universal collection medium (UCM) liquid-based cytology (LBC) (Digene Corp., MD) and the conventional Pap smear in a comparative study, using histologic results as the gold standard. This was a cross-sectional study. Conventional Pap smears and UCM LBC specimens, obtained from women in a low socioeconomic outpatient population referred to a tertiary center for gynecologic care, were compared. For the purpose of this study, when cervical specimens were collected for cytology, all women underwent colposcopy and biopsy was done if a cervical abnormality was observed. Cytologic evaluation of UCM LBC and conventional Pap smears were carried out separately, masked to the results of the other method. Agreement beyond chance between the two cytologic methods was ascertained by means of the unweighted kappa statistic. Sensitivity, specificity, and predictive values with 95% confidence intervals were calculated for both methods. McNemar's test was used to determine the level of association between the two cytology procedures. A total of 800 women were evaluated. Assessment of the overall agreement between the two cytologic methods yielded a kappa of 0.777 (P < 0.0001). After adjustment for histologic diagnosis, the computed kappa in each stratum was as follows: normal = 0.733; CIN 1 = 0.631; CIN 2/3 = 0.735; cancer = 0.652. The sensitivity and specificity of UCM LBC for detection of cervical intraepithelial lesions and cancer were 75.3% and 86.4%, respectively, not statistically different from the 81.8% and 85.2% seen with the conventional method. This study demonstrates that the UCM LBC method is as accurate as the conventional Pap smear cytology in detecting cervical intraepithelial lesions and cancer even so the UCM samples were systematically prepared from a second sampling of the cervix.     

Morphological and immunocytochemical diagnosis of thyroiditis: Comparison between conventional and liquid-based cytology

The efficacy of thyroid (FNAB) processed by liquid-based cytology (LBC) in Hashimoto's Thyroiditis (HT) in two reference periods, is evaluated. The morphologic features of 820 cases with both methods and the cyto-histological comparison are analyzed. The diagnosis of hyperplastic nodules (HN) in HT, its mimickers especially in presence of oxyphilic cells and the role of immunocytochemistry (IHC) are studied. 150 cases of HT processed by conventional smear (CS) in 1996-98 and 670 with LBC in 2005-2007,were included. The majority of FNAB were carried out under USguidance and fixed with ethyl alcohol for the CS. LBC material was rinsed in the Cytolit solution, processed according to the manufacturer's recommendations. Among the 150 CS, 83 were HT while 67 were HN in HT; in the second triennium 245 LBC were HT and 425 were HN in HT. In the first period a follow-up (including a second FNA or surgery) was done in 92 cases, in the second period in 116. In the surgical group 97.1% in the first period were benign (all HT and 34/36 HN) and 2.8% malignant(all HN). In 2005-2007, 94% were benign (15 HT and 45/49 HN) and 6%malignant. Thirty HN from the second triennium had ICC for HBME-1 and Galectin-3 resulting negative in 93.5%. Among these cases, 10 had a benign histology and a concordant negative ICC. LBC can be used as a valid method for HT, especially for the possible application of ICC to HN, and it allows a correct preoperative selection of lesions     

Dutch solutions for liquid-based cytology: analysis of unsatisfactory slides and HPV testing of equivocal cytology

The liquid-based techniques to obtain microscopy slides for cervical screening have replaced conventional smears almost completely in the USA, but not in all European countries. The decision making process to use liquid-based cytology (LBC) for nationwide screening programs depends on the health system. In a pilot study of over 7,000 screenees, we analyzed the unsatisfactory LBC slides and tested the equivocal cytologies for HPV by using the LiPA test. For comparison over 48,000 conventional screening data were used. Compared to conventional smears, the LBC slides were highly cellular, the state of fixation was much better, and obscuring blood did not exist. The unsatisfactory rate showed an increase from 262/100,000 (conventional smears) to 357/100,000 (LBC slides) due to too thick, undiagnosable epithelial fragments on the LBC slides. HPV testing of the equivocal cytology leads to a better patient management and less unnecessary referrals.     

Proliferation patterns of cervical cells as visualized in Leiden liquid cytology slides

The Leiden liquid-based cytology method for the preparation of optimal cytological slides is reported. In such slides, the proliferation pattern of cervical cells can be visualized in detail. Cervical smears and suspension preparations of 665 consecutive unselected patients received in 2003 were studied. Of the 665 patients, 26 (10 normal, 10 with cervical atrophy, 5 with mild dysplasia, and 1 carcinoma in situ) were selected. After using the Thermo Shandon Papspin, the wet slides were placed on a hot plate and dried for 30 min. Proliferation of the cervical cells was visualized in brown by staining the cells for MiB-1 antigen, and nuclear DNA in blue by a standardized short staining with hematoxylin. We found excellent high-resolution demonstrability of cell cycle-related MiB-1 distribution in the well-flattened nuclei. The phase of the cell cycle could be deduced from brown-blue staining patterns. There was a significant increase of MiB-1-positive cell yield related to progression in the degree of pathology.     

Comparison of conventional Papanicolaou smears and fluid-based, thin-layer cytology with colposcopic biopsy control in central Italy: a consecutive sampling study of 461 cases

The aim of this study was to compare the cytologic diagnosis and specimen adequacy of conventional Papanicolaou (CP) and fluid-based, thin-layer [ThinPrep (TP), Cytyc, Boxborough, MA] cervical cytology in a population from central Italy. CP and TP samples were collected simultaneously using a consecutive sampling method on women presenting for cervical screening. Colposcopy was performed as clinically indicated, and biopsy results were compared with cytologic diagnoses. Among the 461 patients included in the study, 413 were negative at both CP and TP, 9 had unsatisfactory results at both tests and 39 patients presented abnormal results at CP, TP or both. Cohen's Kappa was 0.77 showing good agreement between CP and TP test results. Histological data were available for 20 (51.28%) of the 39 patients with at least one positive test. Among the 13 patients with HSIL at histology, 7 had HSIL at CP (sensitivity 53.85%) and 5 at TP (sensitivity 38.46%). For all three patients with squamous cell carcinoma (SCC) at histology, CP and TP had shown the same diagnosis (sensitivity 100%). The positive predictive values were 33.33% for CP and 25.0% for TP regarding the LSIL diagnosis and 100% for both CP and TP regarding HSIL and SCC diagnoses. Our results may be influenced by the consecutive sampling procedure.     

Conventional and liquid-based cervicovaginal cytology: a comparison study with clinical and histologic follow-up

Increased rates of squamous intraepithelial lesion (SIL) diagnosis with liquid-based cervicovaginal cytology (CVC) methods are well documented. This retrospective study compares the ability of the ThinPrep Pap Test (TP) and the conventional Pap smear (CP) to detect biopsy-proven SIL and to exclude nonneoplastic disease. All CVC reports from January 1999 through December 2000 from seven community Family Medicine clinics affiliated with the University of Nebraska were reviewed. For women with at least one CVC diagnosis of epithelial cell abnormality (ECA), follow-up histology, cytology, and clinical data were obtained. Statistical analysis was performed using the chi-square method. SIL was diagnosed in 166 of 3,286 patients by TP (5.1%) and in 169 of 4,872 patients by CP (3.5%) (P < 0.001); 32 of the TP diagnoses (1.0%) and 34 of the CP diagnoses (0.7%) were high-grade SIL (HSIL). Atypical squamous or glandular cells of undetermined significance (ASCUS/AGUS) was the most severe abnormality diagnosed by TP in 218 patients (6.6%) and by CP in 279 patients (5.7%). Follow-up histology data on CVC SIL diagnoses showed evidence of cervical intraepithelial neoplasia in 94 patients screened by TP (2.9%) and in 79 patients screened by CP (1.6%) (P < 0.001); the biopsy diagnoses were CIN 2 or CIN 3 in 34 patients in the TP group (1.0%) and in 28 patients in the CP group (0.6%) (P < 0.025). Follow-up of patients in whom the first ECA CVC diagnosis was ASCUS or AGUS disclosed a positive predictive value for CIN of 22.8% for TP ASCUS/AGUS diagnoses and 11.9% for CP ASCUS/AGUS diagnoses (P < 0.005). In this population, TP was significantly better than CP in detecting biopsy- proven disease and in screening out benign abnormalities.     

Rapid prescreen of cervical liquid-based cytology preparations: results of a study in an academic medical center

A rapid prescreening or rapid rescreening method for quality assurance in cervical cytology has been used in Europe and in Canada but has not been accepted in the United States. The rapid prescreen method was tested in a cytology laboratory that serves an academic medical center with a high‐risk population for cervical cancer. For a period of 3 months, a tray of 20 sequentially numbered Surepath? liquid‐based preparations, randomly selected from the cervical cytology daily workload, were each prescreened in a random fashion for 1 minute. Experienced cytotechnologists performed the rapid prescreen. Results were recorded as negative, further review needed, or epithelial cell abnormality, category specified. The 20 cervical cytology preparations were then replaced in their same position in the daily workload for routine screening performed by another cytotechnologist. Final interpretation was by a cytopathologist as requested or required by Clinical Laboratory Improvement Amendments of 1988. The rapid prescreen data was tabulated and compared with data for a similar time period using the laboratory's normal quality assurance program. Seven hundred and twelve cases underwent rapid prescreen. Six hundred and forty‐two were interpreted as negative. Twenty‐six cases were interpreted as low‐grade squamous intraepithelial lesion (LGSIL) or higher. Forty‐four cases were classified as needing further review. For the 642 negative cases by rapid prescreening, routine screening reported 537 as negative and 105 as either abnormal or needed cytopathologist review. The error rate for the rapid prescreen is 50 of 712 (7.0%); for LGSIL and above 19 of 712 (2.6%). Of the 105 abnormal cases or those submitted for cytopathologist review, 31 were interpreted as atypical squamous cells of undermined significance (ASCUS), 41 cases as reactive/repair, 17 as LGSIL, 4 as unsatisfactory, 1 as atypical squamous cells, cannot rule out high‐grade squamous intraepithelial lesion (ASC‐H), 8 as the presence of endometrial cells in a women aged >40, 1 as malignant melanoma, and 2 as within normal limits with the presence of Actinomyces. The laboratory's routine quality assurance program selects cases, 10% of initially interpreted negative cases plus any gynecologic cytology on patients with a prior abnormal cervical cytology, or history of cervical epithelial cell abnormality. This quality assurance program averages 29% of cases, 4,045 of a total of 13,767, in 2008. Thirty‐seven (0.9%) cases were detected in this rescreen (ASCUS, 16 cases; LGSIL, 13 cases; 1 high‐grade squamous intraepithelial lesion; 4 ASC‐H; and 3 atypical glandular cells of undetermined significance). Eliminating ASCUS cases, eight significant cases were detected, with an error rate of 0.2%. In this cytology laboratory, the rapid prescreen did not prove as reliable as routine quality assurance program for cervical cytology cases. Diagn. Cytopathol. 2012. © 2010 Wiley Periodicals, Inc.     

Cost-effectiveness of liquid-based cytology with or without hybrid-capture II HPV test compared with conventional Pap smears: a study by the French Society of Clinical Cytology

Many articles concerning conventional Pap smears, ThinPrep liquid-based cytology (LBC) and Hybrid-Capture II HPV test (HC II) have been published. This study, carried out by the French Society of Clinical Cytology, may be conspicuous for several reasons: it was financially independent; it compared the efficiency of the conventional Pap smear and LBC, of the conventional Pap smear and HC II, and included an economic study based on real costs; for all the women, a "gold standard" reference method, colposcopy, was available and biopsies were performed whenever a lesion was detected; The conventional Pap smear, the LBC (split-sample technique), the colposcopy, and the biopsies were done at the same time. This study included 2,585 women shared into two groups: a group A of a high-risk population, a group B of a screening population. The statistical analysis of the results showed that conventional Pap smears consistently had superior or equivalent sensitivity and specificity than LBC for the lesions at threshold CIN-I (Cervical Intraepithelial Neoplasia) or CIN-II or higher. It underlined the low specificity of the HC II. Finally, the LBC mean cost was never covered by the Social Security tariff.     

Comparative image analysis of conventional and thin‐layer preparations in endometrial cytology

We evaluated the differences in cytologic findings between conventional and thin‐layer preparations in endometrial cytology to introduce the thin‐layer method into routine cytology. Eighty patients who had undergone endometrial cytology and biopsy on the same day were selected and we compared the cytological findings between conventional‐ and thin‐layer preparations (TLP) in endometrial cytology. The numbers of neutrophils and cell clusters in the thin‐layer method were lower than those in the conventional smear (CSS) method. The average number of neutrophils in endometrioid adenocarcinoma was significantly higher than that in normal morphology endometrium and endometrial hyperplasia. Regarding the shape of the cell clusters, ball‐like patterns and round‐edged cell clusters were not identified in CSS. The average number of clusters in CSS was significantly greater than that using the TLP. The average of the nuclear area in CSS was significantly larger than that using the TLP, indicating that the nuclear areas in CSS were more uneven than that using the TLP. In the future, it is expected that liquid‐based cytology will be applied to the cytological diagnosis of a variety of lesions. The influence on cells due to fixation is considerable in liquid‐based preparations. Therefore, if we strive to pick up the differences between CSS and TLP of endometrial samples, the diagnostic accuracy of the latter could be improved. Diagn. Cytopathol. 2013. © 2012 Wiley Periodicals, Inc.     

Results of an Australian trial using SurePath liquid‐based cervical cytology with Focalpoint computer‐assisted screening technology

BD FocalPoint GS? computer‐assisted screening of BD SurePath® liquid‐based cervical cytology slides (SP + FP) was compared with screening an accompanying conventional cervical Papanicolaou (Pap) smear (CON) in a split sample trial of 2,198 routine specimens. The rate of unsatisfactory specimens in the SP + FP arm was 0.2% compared with 4.1% in the conventional Pap smear, a significant reduction. There was no statistically significant difference between SP + FP and CON for the detection of histologically confirmed high‐grade (HG) lesions in the routine split sample specimens (n = 9). To further test the sensitivity of SP + FP for HG lesions, 38 SurePath slides from confirmed HG cases, without an accompanying CON, were interpolated among the routine smears. In every one of the 47 confirmed HG cases, either HG cells were present in the microscope fields selected by FocalPointGS? for review by the screening cytologist (46 of 47), or full screening of the slide was indicated by the FocalPointGS? (1 of 47), confirming the effectiveness of SP + FP technology for primary screening. In a small number of cases, the screening cytologist did not recognize the abnormality even though on review HG cells were present in fields selected by FocalPointGS?. The overall detection rate was 93% for HG squamous lesions; 89% for known HG endocervical glandular lesions; and 91% for known endometrial carcinoma. In conclusion, the SP + FP detected 100% of HG abnormalities in the trial set; significantly reduced the rate of unsatisfactory specimens; and improved the overall screening rate of detection of HG abnormalities particularly of glandular lesions when compared with other screening technologies. Diagn. Cytopathol. 2012. © 2011 Wiley Periodicals, Inc.     

Automation in cytology: A survey conducted by the New Technology Task Force,Papanicolaou Society of Cytopathology

Despite the overwhelming interest in the development of several computer based technologies in the last several years, the role of automation in cytology has remained controversial. The potential of these technologies in the reduction of false negative results in pap smears is well recognized. However, there is still remarkable confusion as how to incorporate automation in the routine practice of cytology. This prompted the New Technology Task Force of the George Papanicolaou Society of Cytopathology to design a survey to seek the opinion of those engaged in cervicovaginal cytology screening regarding the value of automation in cytology. In 1996, a ten question survey was sent to 1800 cytopathology laboratories throughout the nation. The response rate was 23% (416/1800). The responders represented laboratories varying from those with less than 5,000 pap smears to those with over 100,000 cases per year. The majority of the responders did not believe that automation is essential for cervicovaginal cytology. This was evidenced by the fact that only 12% of the laboratories were engaged in automated cytology and predominantly used it for quality control measures. The inability of small laboratories to absorb the extra expense involved in the integration of automated cytology in their practice, particularly in the current era of managed care was a major concern. There was also concern about the potential for compromise of patient care by the drive for corporate profits and the dissemination of wrong information to the public and physicians. Suggestions most frequently proposed included appropriate patient and physician education about the merits and pitfalls of the pap smear, and also endorsing an affordable universal fee for pap smears. Rescreening for all pap smears, reassessing the benefits of automation in cytology and development of the standards were other proposals. Partnership with larger cytology laboratories, creation of “cytology consortiums” with shared resources to provide regionalized automated rescreening services were also strongly suggested. This survey clearly indicates the need for further evaluation of automation in cytopathology and a focused attention to various issues surrounding cervicovaginal cytology screening. Diagn. Cytopathol. 1998; 18:47–55. © 1998 Wiley-Liss, Inc.     

Utility of ProExC and IMP3 immunocytochemical staining in atypical glandular cells of undetermined significance in liquid‐based cervical cytology

The diagnosis of atypical glandular cells of undetermined significance (AGUS) in liquid‐based cervical cytology specimens shows significant underlying pathology in only 30% of cases, while the remaining cases are found to be benign (reactive, reparative/metaplastic). Previous studies have reported positive ProExC and IMP3 staining in neoplastic glandular lesions of the uterine cervix and corpus. We present our experience with the utility of these markers in the evaluation of AGUS cases in liquid‐based cervical cytology. The case cohort included 34 cases diagnosed as AGUS. ProExC and IMP3 immunocytochemical (ICC) stains were performed on ThinPrep® slides and the results correlated with subsequent biopsy findings. Positive expression was classified as strong diffuse nuclear immunostaining for ProExC and granular cytoplasmic for IMP3. The presence of AGUS cells on the ICC stained slides was confirmed in all cases. IMP3 was positive in 80% of glandular neoplasms and negative in 93% non‐glandular lesions/cases negative for squamous intraepithelial lesion (SIL). ProExC was positive in 60% of glandular neoplasms and negative in 83% non‐glandular lesions/cases negative for SIL. When used as a panel (ProExC + IMP3), at least one stain was positive in 100% of glandular neoplasm cases and they were both negative in 83% of non‐glandular lesions/cases negative for SIL. Based on this study, both ProExC and IMP3, when used as an immuno panel, can predict the presence of glandular lesions on subsequent biopsies and can serve as an aid in the diagnosis and management of AGUS cases. Diagn. Cytopathol. 2014;42:375–379. © 2013 Wiley Periodicals, Inc.     

Liquid‐based cytology using cytorich red/tripath is diagnostically equivalent to conventional smears for bronchial washings and brushings and reduces the cost

A split sample study was performed on 109 bronchial brushings and washings and evaluated from conventional slides (CS) and CytoRich Red/Tripath preparations (CRR/Tripath). Unassessable bronchial washings were significantly more frequent in CS (5 vs. 0), but as all brushings were assessable with both methods, no overall diagnostic advantage was found. CS and CRR/Tripath gave discordant diagnoses in one case with a final benign diagnosis and in six cases with final malignant diagnoses. In the benign case, atypia was assessed in CS. In the malignant cases, suspected malignancy was found in one CRR/Tripath and one CS, atypia vs. benign assessment was also balanced, with three atypias in CRR/Tripath and two in CS. The better preserved cells in CRR/Tripath facilitated correct diagnosis in some cases, but might also lead to false positive diagnoses. In small cell carcinomas diagnostic hints such as smearing and moulding were less pronounced in CRR/Tripath but this did not affect the diagnostic accuracy. Overall, the diagnostic performance with CRR/Tripath was at least as good as with conventional slides, although statistically no difference could be seen. The number of slides and screening time, and thereby cost was significantly reduced with CRR/Tripath, thus the liquid‐based method is preferred for bronchial washings and brushings. Diagn. Cytopathol. 2013;41:876–884.© 2013 Wiley Periodicals, Inc.     

A comparative study between conventional and liquid‐based cytology in screening for anal intraepithelial lesions in HIV‐positive patients

Anal intraepithelial neoplasia (AIN) is associated with HPV infection and can be detected by cytological screening. While conventional exfoliative cytology (CC) is a low‐cost and nonaggressive method, liquid‐based cytology (LBC) tends to give clearer readings. Although studies of the efficacy of anal cancer screening methods would be of great importance for groups at high risk for AIN, few such studies have been conducted. The aim of the present study was to assess the concordance of CC and LBC in diagnosing anal pre‐neoplastic lesions, and to compare cytological results with anoscopy, histopathological, and molecular biology findings. Comparative study involving 33 HIV‐positive patients, who underwent anoscopy and biopsy of suspected lesions. Concordance between the two cytology methods was calculated, as were the associations between cytology results and findings from other screening methods. A total of 54.5% of cases were considered AIN‐negative by CC and LBC, and concordance between the two methods was statistically significant (P < 0.05). Anoscopy was negative in 15 of the 18 CC‐ and LBC‐negative cases. CC identified 75% of patients with positive biopsy, while LBC identified 85.71% of these patients. Molecular biology results showed that patients with LSIL tested positive for the highest number of HPV subtypes. The associations between positive biopsy and high grade HPV, HPV 16, and multiple HPV infections were not statistically significant. Conventional and liquid‐based cytology are equally effective in screening for anal preneoplastic lesions. Diagn. Cytopathol. 2014;42:840–845. © 2014 Wiley Periodicals, Inc.     

Cost-effectiveness of monolayers and human papillomavirus testing compared to that of conventional Papanicolaou smears for cervical cancer screening: protocol of the study of the French Society of Clinical Cytology

The French Society of Clinical Cytology is conducting a study to compare the cost-effectiveness of monolayers and human papillomavirus (HPV) testing with that of conventional Papanicolaou (Pap) smears for cervical cancer screening. The protocol of this study is presented. It includes 3,000 women who will be evaluated by the three methods (conventional Pap smears, or monolayers with or without HPV testing) and by the reference method: colposcopy followed, in cases with abnormalities, by cervical biopsy. Efficacy or performance of the methods will be compared on the basis of sensitivity. Cost comparisons and cost-effectiveness modeling will be based on the costs associated with methods themselves and also the costs of "false positives." This will require specific collection of data concerning the costs of the three methods, as these costs have not previously been accurately documented. Patient recruiting and data collection started in September 1999 and will be complete in June 2000. The first results are expected to be available in spring 2001.     

Effectiveness of the SurePath liquid-based Pap test in automated screening and in detection of HSIL

We have shown that SurePath when compared to conventional Paps fails to increase HSIL detection. In this study, assessment of test performance characteristics for the FocalPoint showed that sensitivity was 96% when manual screening was used as the "gold standard." When cervical biopsy, however, was used as the "gold standard" FocalPoint sensitivity decreased to 93%, which was the same as manual screening. Examination of the FocalPoint "no further review" cases showed that 4/296 were SIL. To understand better the implication of an SIL diagnosis, cervical biopsies generated from SurePath and conventional Paps were compared. Conventional Paps diagnosed as LSIL had a biopsy LSIL:HSIL ratio of 3.1/1, while SurePath Paps had a biopsy LSIL:HSIL ratio of 1.5/1. These results indicate that when cervical biopsy is used as the "gold standard," FocalPoint and manual screening of SurePath Paps have similar test performance but that the FocalPoint can fail to detect LSIL. Importantly, LSIL in a SurePath Pap is 1.6 times more likely to be HSIL on biopsy than LSIL in a conventional Pap.     

Overexpression of p16INK4A in liquid-based specimens (SurePath) as marker of cervical dysplasia and neoplasia

Human papillomavirus (HPV) is recognized as a causal agent for cervical carcinomas. Assimilation of HPV oncogenes E6 and E7 into the host DNA promotes upregulation of cyclin dependent kinase inhibitor (CDKI) p16(INK4A), detectable by monoclonal antibody in the developing cervical cancer cells. The aim of this study was to 1) develop a protocol for p16(INK4A) immunocytochemical staining on SurePath preparations, and 2) determine its utility as an HPV marker on a spectrum of cervical reactive and neoplastic lesions. Seventy-two specimens consisting of 28 nonneoplastic/nondysplastic cases (NN), one reactive glandular cells (RGC), 27 low-grade squamous intraepithelial lesions (LSIL), 10 high-grade squamous intraepithelial lesions (HSIL), one squamous cell carcinoma (SCCA), four atypical glandular cells (AGUS), and two adenocarcinomas (ADCA) were reprepped by SurePath and antibody to p16(INK4A) applied at 1:100 dilution using the Dako Envision + System on the Dako Autostainer. Expression of p16(INK4A) within the nucleus principally and cytoplasm of at least 10-15 cells was considered positive. All initial Papanicolaou-stained discrepant cases (p16(INK4A) positivity of NN and RGC cases and lack of reactivity in LSIL, HSIL, and AGUS) were reviewed. Nine of ten (90%) HSIL, one (100%) SCCA, 21/27 (78%) LSIL, and some reactive and inflammatory specimens demonstrated the presence of p16(INK4A). Reevaluation of discrepant cases revealed that several were underinterpreted (four NN were LSIL, one RGC was AGUS) or overinterpreted (one LSIL was NN). Following reassessment, false-positive staining was present in only 1/25 (1.4%) NN. Six of 30 (20%) LSIL lacked p16(INK4A) positivity. One of 10 (10%) HSIL had no staining. Two of four AGUS did not react with p16(INK4A) antibody. Both SCCA (1) and ADCA (2) had positive expression. This study confirms the intimate relationship between p16(INK4A) and HPV cytopathic effect. The p16(INK4A) immunocytochemical stain can be applied to liquid-based cervical preparations. This technique offers a more objective approach to deciphering "gray areas" of gynecologic cytopathology.     

PREPmate automated processor: comparison of automated and manual methods of liquid-based gynecologic sample preparation

The development of procedures for fully automated processing of liquid-based gynecologic samples has been the focus of considerable interest to the cytology laboratory. Liquid-based collection and processing technology has been shown to improve sample adequacy, resulting in an overall improvement in quality of sample preparations. PREPmate, an accessory to the PrepStain slide processor, automates the initial enrichment process of mixing and dispensing the specimen over a density gradient. This report describes a study evaluating cellularity and diagnostic reproducibility in SurePath samples processed using the PREPmate accessory compared to samples processed using a manual technique. Samples processed using the PREPmate accessory contained 8.3% more squamous cells. Exact diagnostic reproducibility between preparation types was 83.3%; when considering negative vs. abnormal (ASCUS+), in adequate samples, reproducibility was 100%.