Liquid-based Cervical Cytology Specimens Research Articles

Evaluation of dual p16/Ki-67 immunostaining on anal cytology specimens.

Dual immunostaining for p16/Ki67 is FDA-approved for use on liquid-based cervical cytology specimens; however, the utility of dual staining in anal cytology especially for ASCUS risk stratification is not well established. We investigated dual staining performance on anal cytology specimens and correlated with subsequent cytologic interpretation, high-risk HPV status, and anal biopsy results. Dual staining for p16/Ki-67 was performed on all liquid-based anal cytology specimens from December 2021 to June 2022 (n = 43). Three patients had high grade squamous intraepithelial lesion (HSIL/AIN2-3) on biopsy; dual staining was positive in all three cases. All HR-HPV negative cases were negative for dual staining. Among the 12 ASCUS samples with subsequent anal biopsy results all also had HR-HPV testing. Due to small sample size of cases with squamous intraepithelial lesion (SIL) diagnosed on biopsy, the sensitivity and positive predictive value was not calculated. However, the specificity and negative predictive value of p16/Ki-67 dual staining for SIL of any grade on biopsy were 1 (95% CI: 0.66-1) and 0.9 (95% CI: 0.65-0.97) respectively, whereas the specificity and negative predictive value of HR-HPV testing for SIL of any grade on biopsy were 0.44 (95% CI: 0.14-0.79) and 0.8 (95% CI: 0.41-0.96) respectively. Dual p16/Ki-67 staining indicates transforming HPV infection and could help serve as an ancillary test for risk stratification for atypical anal cytology specimens. Among ASCUS samples, dual staining was specific for SIL of any grade with a high negative predictive value and therefore could be useful in clinical practices with limited availability for follow-up care.

A piezoelectric immunosensor for the rapid detection of p16INK4a expression in liquid-based cervical cytology specimens

A piezoelectric immunosensor was developed for the detection of p16INK4a, a protein that is expressed in liquid-based cervical cytology specimens and has been proved to be linked cervical cancer. Mouse polyclonal antibody against p16INK4a was bound onto the piezoelectric crystal surface in an ordered orientation using protein A. The antibody on the crystal could specifically adsorb p16INK4a and changes in the mass of the crystal would led to frequency shift. The supernatants from different clinical samples, including various cervical lesions, containing different concentration of p16INK4a were used for testing. Under the optimized conditions, the frequency shifts were linearly dependent on the antigen concentration in the range of 50–1200ngmL−1. The device showed a short analysis time (less than 30min), good reproducibility (could be reused six times without any detectable loss of activity), stability (the immunosensor was stable for more than eight weeks when stored over silica gel blue at 4°C), and specificity. Attempts have been made to use the sensor for the initial screening for cervical lesion in cervical cytology specimen, and the relationship between cytologic and histologic findings, p16INK4a expression in various cervical lesions, including carcinoma been investigated.

The p16INK4a Antibody Immobilization Method for Immonosensor Application.

The p16INK4a is a protein that expressed in Liquid-based cervical cytology specimens and has been proved link to cervical cancer. The p16INK4a could be detection by piezoelectric immunosensor and the immobilization of the p16INK4a antibody influence the sensitivity of the piezoelectric immunosensor. 5μL mouse polyclonal antibody against p16INK4a was bound onto the surface of immonosensor through two methods. (directly immobilized method; protein A method). Absorb of the p16INK4a antibody on the surface of immonosensor caused a shift in the resonant frequency of the immunosensor and The frequency changes recorded showed a better reproducibility. The activity of the immobilization antibody with the directly method and protein A method was tested with p16INK4a antigen. The resonant frequency for different antibody immobilization methods were different, and the sensitivity for p16INK4a detection also different. The protein A method was found to be much more better than the directly method for the immobilization of the p16INK4A antibody on the gold electrode of the quartz crystal for cervical lesion detection. The Protein A method created more reproducible and stable immobilization antibody layers with p16INK4A antigen.

Genotypes and prevalence of HPV single and multiple concurrent infections in women with HSIL

The contribution of human papillomavirus (HPV) types to the carcinogenesis of cervical cancer has been established for a long time. However, the role of phylogenetically related and rare variants remains uncertain, as well as the influence of concurrent multiple HPV genotypes infection. We aimed at studying the prevalence of several HPV genotypes infecting women with single versus concurrent multiple HPV genotypes infection with a HSIL diagnosis in a cervical cytology. We conducted a cross-sectional study using Thin-Prep(®) liquid-based cervical cytology specimens with the diagnosis of high-grade squamous intraepithelial lesion (HSIL), in which HPV genotype was sequentially tested. Genotypes were determined with a PapilloCheck(®) system, a DNA-Chip for the type-specific identification of 18 high-risk and six low-risk types of HPV. Of the total study population, 176 cases had a diagnosis of HSIL and positive HPV genotyping result, being HPV16 the most prevalent genotype (48.86%; 95%CI: 41.58-56.19) followed by HPV31 (14.20%; 95%CI: 9.75-20.18). Concurrent multiple HPV genotypes were detected in 36.93% (95%CI: 30.15-44.27) of the patients. The prevalence of the 10 most common HPV genotypes detected varied significantly according to the presence of single vs. concurrent multiple HPV genotypes (P = 0.022). Moreover, women with concurrent multiple HPV genotypes were on average 3.53 (95%CI: 0.43-6.64) years younger than women with single genotype infection. Our results suggest that women with multiple genotype HPV infection differ in terms of age and distribution of the most prevalent HPV genotypes. Additionally, we provide further evidence of the predominance of HPV16 in HSIL lesions of the uterine cervix.

L1 C-terminal common sequence polypeptide antibody in detecting human papillomavirus in liquid-based cervical cytology specimens

Objective To determine whether the polypeptide antibody induced by human papillomavirus( HPV) L1 Cterminal conserved sequence polypeptide can be used to detect HPV in cervical exfoliated cell specimens. Methods Cervical exfoliated cell specimens were collected and divided into two groups: one was subjected to nested polymerase chain reaction( Nest-PCR) for detection of HPV DNA,and the other was subjected to sandwich enzyme-linked immunosorbent assay( ELISA),in which rabbit antipolypeptide purified antibody and mice anti-polypeptide antiserum were used to determine HPV L1 in specimens. The detection rates of HPV were compared between the two groups. Part of the specimens were used for immunocytochemistry detection. Results Nest-PCR with primer MY09 /11 detected 30 HPV DNA positive specimens out of a total of 296 ones,with a detection rate of 11. 15%. Then the HPV DNA negative specimens were further amplified with primer GP5 + /6 +,and 51( 21. 34%) more positive cases wereidentified,making the total detection rate of Nest-PCR being 30. 11%( totally 81 cases). ELISA identified 91 positive cases out of263 specimens,with a detection rate of 34. 60%,which was not significantly different from that by Nest-PCR. Immunocytochemistry detection results displayed that the polypeptide antibody could specifically reveal cervical exfoliated cells infected with HPV.Conclusion The HPV L1 polypeptide antibody can satisfactorily detect the HPV infection in cervical exfoliated cells,indicating that HPV L1 polypeptide antibody has great potential in developing HPV detection kit for cervical cancer preventive screening.

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.

Cytopathologic Features of Molluscum Contagiosum in a Liquid-Based Cervical Cytology Specimen

Cytopathologic Features of Molluscum Contagiosum in a Liquid-Based Cervical Cytology Specimen

Promoter methylation of DAPK1, RAR-β and MGMT in exfoliated cervical cytology and its clinical application

To assess the correlation of promoter methylation of DAPK1, RAR-β and MGMT with cervical lesions from cytology to histology, and to reveal the clinical value of DNA methylation in diagnosis of cervical intraepithelial neoplasia (CIN). A total of 103 random-selected cervical samples were collected from residual liquid-based cytology specimens after clinical use in cytopathological diagnosis in outpatient clinic of obstetrics and gynecology, Peking Union Medical Collage Hospital from March 2010 to October 2010. Informed consent was obtained from each woman before the initiation of the study. The methylation sensitive-high resolution melt (MS-HRM) assay was used to evaluate promoter methylation of three genes (DAPK1, RAR-β and MGMT) in 103 biopsy-confirmed liquid-based cervical cytology samples. Methylation levels and high-risk HPV DNA loading (HCII values) were analyzed in relation to both cytological and histological diagnosis. The methylation level of all three genes showed significant difference among the different cytological groups (P = 0.000, 0.011 and 0.002, respectively). The methylation level of DAPK1 and RAR-β showed significant difference among the different histological groups (P = 0.000 and 0.021), while there was no significant difference for MGMT. DAPK1 methylation levels was 1.47% in the CINII/high-grade precancerous lesions group, and 20.98% in the normal/CINI groups (P = 0.000), but there was no significant difference between CINII/high-grade precancerous lesions and normal/CINI groups for RAR-β and MGMT. The combination of DAPK1/HR-HPV loading showed a sensitivity of 0.825 and an area under the receiver operating characteristic curve (ROC) curve (AUC) of 0.695 as diagnostic methods for detecting CINII/high-grade precancerous lesions. DNA methylation such as DAPK1 and RAR-β, in combination with HR-HPV detection, may serve as biomarkers to detect CINII/high-grade precancerous lesions. Detection of methylated DNA from liquid-based cervical cytology specimens is technically feasible with the MS-HRM assay.

ProExC Triage of Atypical Glandular Cells on Liquid-Based Cervical Cytology Specimens

Morphologic distinction between atypical glandular cells not otherwise specified (AGC-NOS) and AGC-favor neoplasia (AGC-FN) can be difficult. Distinction between these entities is important as the American Society for Colposcopy and Cervical Pathology 2006 consensus guidelines state that management of AGC-NOS differs from that of AGC-FN. The objective of this study was to determine the potential role of ProExC immunocytochemical triage of AGC-NOS. Cytopathology records from a pathology practice were reviewed from January 2006 to December 2009 to identify AGC-NOS liquid-based Pap smears with subsequent biopsy correlation. Archival slides were examined, and ProExC immunocytochemistry was performed. The AGC groups were assessed for nuclear staining, and results were correlated with subsequent biopsy findings. Twenty-eight AGC-NOS cases with biopsy correlation were identified: 13 with subsequent high-grade neoplastic or malignant (positive) diagnoses and 15 with benign diagnoses. Of 13 AGC-NOS cases with positive diagnosis, 10 were ProExC-positive and 3 were ProExC-negative (metastatic tumors from distant sites). Of 15 AGC cases with benign follow-up, 13 were ProExC-negative and 2 were ProExC-positive (sensitivity, 77%; specificity, 87%). For patients with cervical intraepithelial neoplasia or carcinoma originating from the female genital tract, 100% (10/10) were ProExC-positive (sensitivity, 100%; specificity, 87%). Results suggest that ProExC-positive AGC-NOS may be classified as AGC-FN. Although positive immunocytochemical staining for ProExC requires management similar to AGC-FN, negative staining does not rule out malignancy such as metastatic tumor. Management for ProExC-negative AGC-NOS cases should proceed according to the current guidelines for AGC-NOS.

Quantification of intracellular HPV E6/E7 mRNA expression increases the specificity and positive predictive value of cervical cancer screening compared to HPV DNA

Objectives Current methods for HPV screening rely on the detection of L1 DNA from high risk genotypes (HRHPV). These assays have very high negative predictive values (~ 99%), however, the specificity and positive predictive value of HPV DNA tests for pre-cancerous and cancerous lesions (CIN 2+) is less than 50%. The purpose of this study was to compare HPV DNA with intracellular HPV E6, E7 mRNA quantification in an effort to improve the performance of cervical cancer screening. Methods Liquid-based cervical cytology specimens collected in either PreservCyt or SurePath were processed for routing cytology, HPV HRDNA detection by Hybrid Capture 2 and HPV E6, E7 mRNA quantification in cells using the same sample. We analyzed a total of 2049 samples including 73 with CIN 2, CIN 3, or squamous cell carcinoma by biopsy and 1694 samples from women with normal cytology. Results The positive predictive value of HPV E6, E7 mRNA quantification in cells for CIN2+ was 78% which was greater than HPV DNA alone (43%). The specificity of HPV E6, E7 mRNA quantification was 96% based on normal cytology compared to 82% for HC2 while the specificity of HPV E6, E7 mRNA quantification based on CIN 2− histology was 85% compared to 35% for HC2. Conclusions With similar sensitivity and greater specificity/positive predictive value, HPV E6, E7 mRNA quantification in cells is an improvement over HPV DNA for cervical cancer screening.

Outcomes of cervical liquid‐based cytology suggesting a glandular abnormality

To ascertain the positive predictive value of both ?glandular neoplasia (national standard code 6) and borderline change (national standard code 8) in glandular cells in liquid-based cervical cytology specimens in Cardiff and Vale NHS Trust and to outline the histological outcomes of these cases. Eighty-nine liquid-based (Surepath) cervical cytology cases were retrospectively identified from a 2-year period (January 2005 to December 2006) and correlated with histopathological diagnoses. Initial punch biopsy histology revealed 18 cases (21%) of cervical glandular intraepithelial neoplasia (CGIN). A further nine cases (10%) of CGIN were identified following local excision or hysterectomy. Ten cases of invasive malignancy were identified: four endocervical adenocarcinomas (all node negative, TNM stage T1b1), five endometrial adenocarcinomas and one squamous cell carcinoma. There were 10 with high-grade cervical intraepithelial neoplasia (CIN) alone. Women diagnosed with endometrial malignancy presented later with an average age of 64.6 years compared with 34.9 years for endocervical lesions. Taking high-grade CIN or worse as a positive outcome, the overall positive predictive value (PPV) of glandular abnormalities on cytology (both code 6 and 8) was 58.1% [95% confidence interval (CI) 47.8, 68.4]. PPV for borderline change in glandular cells alone was 24.1% (95% CI 8.5, 39.6) and for ?glandular neoplasia alone 75.4% (95% CI 64.3, 86.5). With our interpretation of the classification, women with cytological diagnoses of glandular neoplasia of the cervix should initially be investigated by local resection rather than punch biopsy, and those with borderline change in glandular cells with repeat cytology.

Rapid, high throughput determination of cervical cytology specimen adequacy using a capillary‐based cytometer

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.

Evaluation of candidate methylation markers to detect cervical neoplasia

Objective. Studies of cervical cancer and its immediate precursor, cervical intraepithelial neoplasia 3 (CIN3), have identified genes that often show aberrant DNA methylation and therefore represent candidate early detection markers. We used quantitative PCR assays to evaluate methylation in five candidate genes (TNFRSF10C, DAPK1, SOCS3, HS3ST2 and CDH1) previously demonstrated as methylated in cervical cancer. Methods. In this analysis, we performed methylation assays for the five candidate genes in 45 invasive cervical cancers, 12 histologically normal cervical specimens, and 23 liquid-based cervical cytology specimens confirmed by expert review as unequivocal demonstrating cytologic high-grade squamous intraepithelial lesions, thus representing the counterparts of histologic CIN3. Results. We found hypermethylation of HS3ST2 in 93% of cancer tissues and 70% of cytology specimens interpreted as CIN3; hypermethylation of CDH1 was found in 89% of cancers and 26% of CIN3 cytology specimens. Methylation of either HS3ST2 or CDH1 was observed in 100% of cervical cancer tissues and 83% of CIN3 cytology specimens. None of the five genes showed detectable methylation in normal cervical tissues. Conclusion. Our data support further evaluation of HS3ST2 and CDH1 methylation as potential markers of cervical cancer and its precursor lesions.

Evaluation of quantity and staining pattern of human papillomavirus (HPV)-infected epithelial cells in thin-layer cervical specimens using optimized HPV-CARD assay

Testing for human papillomavirus (HPV) is used in the triage of women with a cervical cytology of atypical squamous cells of undetermined significance (ASCUS). A fluorescent in situ hybridization assay was developed for the detection of HPV using the catalyzed receptor deposition technique (HPV-CARD). In this study, the utility of this assay was tested for the detection of HPV in liquid-based cervical cytology specimens. A total of 195 liquid-based cytology specimens were analyzed using the HPV-CARD assay. The results from the assay were compared with HPV polymerase chain reaction (PCR) and typing results. The number of HPV-infected cells and the staining pattern was correlated with the cytology classification. A 91% concordance between HPV-CARD and PCR was observed for the detection of high-risk HPV viruses. A 78% concordance was observed for specimens that were negative for HPV. In ASCUS, low-grade squamous intraepithelial lesion (LSIL), and high-grade squamous intraepithelial lesion (HSIL) categories, the average number of HPV-positive cells per slide was 19 cells, 127 cells, and 450 cells, respectively. The number of cells with a punctate staining, suggestive of HPV integration, was 21% in ASCUS, 34% in LSIL, and 46% in HSIL specimens. The results of the current study indicate positive correlations between the severity of the disease and the increased overall quantity of HPV-positive epithelial cells in cervical cytology specimens and accumulation of cells with punctate staining suggestive of integrated HPV. In summary, the developed HPV-CARD assay was found to provide novel information regarding the proportion and staining pattern of HPV-infected epithelial cells in different cytologic categories of cervical specimens.

Normal endometrial cells in liquid-based cervical cytology specimens in women aged 40 or older

Objective. The 2001 Bethesda System consensus statement directs the reporting of endometrial cells on cervical cytologic preparations in women aged 40 years or older. Our objective was to assess the significance of endometrial cells on cervical liquid-based cytology (LBC) specimens in this population. Methods. The population included women aged 40 years or older in whom endometrial cells were identified in otherwise normal LBC specimens from December 2001 through June 2005, and who subsequently underwent endometrial sampling within a 12-month period. Patient age, menopausal status, hormonal use, the presence or absence of co-incident symptoms and endometrial sampling results were recorded. Results. Endometrial cells were identified on LBC specimens in 2494 women during the study period, for an incidence of 0.4%. Of these, 370 women underwent endometrial sampling within 12 months of the incident LBC preparation and met inclusion criteria. In asymptomatic premenopausal women, sampled solely due to the presence of endometrial cells, 2.1% were identified with significant endometrial pathology, compared to 2.6% of symptomatic premenopausal women. In menopausal women, only those with symptoms (mainly bleeding) had significant pathology on subsequent sampling (25%); none of the asymptomatic menopausal women were found to have hyperplasia or carcinoma. No significant differences were identified in premenopausal women according to the time of sampling during the menstrual cycle or use of oral contraceptives. Conclusion. Endometrial cells on LBC preparations, even in the absence of symptoms in premenopausal women aged 40 years or older, are associated with significant uterine pathology.

Response to ‘Using a quality control approach to define an ‘‘adequately cellular’’ liquid‐based cervical cytology specimen’

CytopathologyVolume 18, Issue 5 p. 326-326 Response to ‘Using a quality control approach to define an ‘‘adequately cellular’’ liquid-based cervical cytology specimen’ First published: 26 February 2007 https://doi.org/10.1111/j.1365-2303.2007.00418_2.x Edward Duvall and Fiona McQueenRoyal Infirmary of Edinburgh, Edinburgh, UKE-mail: e.duvall@ed.ac.uk Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume18, Issue5October 2007Pages 326-326 RelatedInformation

Using a quality control approach to define an ‘adequately cellular’ liquid‐based cervical cytology specimen

To define a minimum acceptable total squamous cellularity for (ThinPrep) liquid-based cervical cytology (LBC) specimens using quality control techniques. Two hundred LBC preparations were made containing varying numbers (<200) of severely dyskaryotic squamous cells and with varying total cellularities. Ninety-eight per cent of the LBC preparations that were missed by one or more of three cytoscreeners had fewer than 16 abnormal objects (single dyskaryotic cells or clumps of cells) and 87 dyskaryotic cells. The minimum ratio of dyskaryotic to total squamous cells that, in a preparation of 5000 squamous cells has a probability of at least 0.98 that 87 or more dyskaryotic cells will be present is 1:47. Twenty-three preparations diagnosed as abnormal had ratios of dyskaryotic to total squamous cells of between 1:2.5 and 1:4596. There is thus no feasible minimum acceptable squamous cellularity that will give an acceptable probability of detection of all specimen vials containing abnormal cells in the observed proportions. It is suggested that the minimum acceptable cellularity for LBC specimens is set pragmatically by the screening programme to give a feasible percentage of repeat tests.

High-Throughput Cervical Cancer Screening Using Intracellular Human PapillomavirusE6andE7mRNA Quantification by Flow Cytometry

The Papanicolaou (Pap) test, based solely on the morphologic examination of exfoliated cells from the cervix, has reduced deaths due to cervical cancer by 74% in the United States during the past 40 years. During that time, the molecular mechanisms of cervical cancer have largely been elucidated. Taken together, these observations have identified a need for a high-throughput cervical cancer screening assay. We report the development of a high-throughput assay consisting of simultaneous immunophenotyping and ultrasensitive in situ hybridization for human papillomavirus (HPV) E6 and E7 messenger RNA (mRNA). This assay can be performed in less than 3 hours directly from liquid-based cervical cytology specimens. Overall, HPV fluorescence in situ hybridization (FISH) for E6 and E7 mRNA demonstrated 83.3% sensitivity and 91.3% specificity for high-grade squamous intraepithelial lesions compared with the Pap test in 231 liquid-based cytology samples from 2 cohorts. In a subset of these samples, HPV FISH demonstrated higher sensitivity and specificity than Hybrid Capture (Digene, Gaithersburg, MD) for high-risk genotypes.

A Novel Filtration-Based Processing Method of Liquid Cytology Specimens for Human Papillomavirus DNA Testing by Hybrid Capture II

We evaluated a more efficient method of processing liquid-based cervical cytology specimens for human papillomavirus (HPV) DNA testing by Hybrid Capture II (HCII). Aliquots were made from 701 specimens in the following sequence: 4.0, 2.0, 1.0, 0.5, and 1.5 mL. The 4.0-mL aliquot was processed by the standard method (STP), and half of the processed material was tested by HCII. Other aliquots were processed with a new, filtration-based processing method (NPM). The 2.0-mL NPM aliquot had HCII test performance most similar to the STP, ie, similar HCII positivity (P = .4) and good test agreement (κ = 0.85, 95% confidence interval [CI], 0.80–0.89). The 194 cytologic negatives had greater positivity by STP (P = .04) compared with the 2.0-mL aliquot processed by NPM; between-method agreement was modest (κ = 0.54, 95% CI, 0.36–0.72). A lower positive cut point for the 2.0-mL NPM aliquot partially abrogated this minor difference. In 241 specimens diagnosed as low-grade and 31 as high-grade squamous intraepithelial lesions, there were no significant differences in HPV positivity (>85% and 90%, respectively) between STP and NPM. NPM reduces specimen handling and decreases total testing time by approximately 33% without significant losses in HCII test performance.

A Novel Filtration-Based Processing Method of Liquid Cytology Specimens for Human Papillomavirus DNA Testing by Hybrid Capture II

We evaluated a more efficient method of processing liquid-based cervical cytology specimens for human papillomavirus (HPV) DNA testing by Hybrid Capture II (HCII). Aliquots were made from 701 specimens in the following sequence: 4.0, 2.0, 1.0, 0.5, and 1.5 mL. The 4.0-mL aliquot was processed by the standard method (STP), and half of the processed material was tested by HCII. Other aliquots were processed with a new, filtration-based processing method (NPM). The 2.0-mL NPM aliquot had HCII test performance most similar to the STP, ie, similar HCII positivity (P = .4) and good test agreement (kappa = 0.85, 95% confidence interval [CI], 0.80-0.89). The 194 cytologic negatives had greater positivity by STP (P = .04) compared with the 2.0-mL aliquot processed by NPM; between-method agreement was modest (kappa = 0.54, 95% CI, 0.36-0.72). A lower positive cut point for the 2.0-mL NPM aliquot partially abrogated this minor difference. In 241 specimens diagnosed as low-grade and 31 as high-grade squamous intraepithelial lesions, there were no significant differences in HPVpositivity (>85% and 90%, respectively) between STP and NPM. NPM reduces specimen handling and decreases total testing time by approximately 33% without significant losses in HCII test performance.