Human Papillomavirus Genotype Detection and Cytology Using a New Self-Sampling Method.

OBJECTIVE: High-risk human papillomavirus (hrHPV) testing using dry-type self-sampled vaginal specimens is becoming more widespread worldwide due to increased screening uptake. However, for the triage of hrHPV-positive women, a visit to a general practitioner is required for reflex cytology. This study aimed to evaluate the hrHPV detection capability of CellSoft®, a wet-type self-sampling method that also allows for cytology. METHODS: Thirty-eight women aged 20 years and older were included in the study. The women self-sampled using CellSoft® after using an Evalyn® Brush. PCR-based HPV genotyping was performed on both specimens and hrHPV detection results of both devices were compared. Additionally, cytological exam was performed on CellSoft® samples. RESULTS: Overall agreement between self-sampling devices for the detection of hrHPV in CellSoft® and Evalyn Brush was observed in 97.4% (37/38) of participants. More hrHPV genotypes were detected with Evalyn Brush than with CellSoft®. Among the 22 CellSoft® hrHPV-positive cases, 11 (47.6%) were atypical squamous cells of undetermined significance or worse. CONCLUSION: CellSoft® hrHPV genotype detection results were in good agreement with those of Evalyn Brush. CellSoft® provided a sufficient cell volume for HPV testing and cytological evaluation.

A study of the relationship between nuclear contour thickening, nuclear enlargement and human papillomavirus infection in squamous cells.

OBJECTIVE: To investigate the relationship between the morphological features of nuclear enlarged cells and human papillomavirus (HPV) infection in negative for intraepithelial lesion or malignancy (NILM) and atypical squamous cells of undetermined significance (ASC-US). METHODS: In total, 128 Papanicolaou specimens comprising 41 ASC-US cases and 87 NILM cases were examined. Cell morphological analysis was performed using both area ratio (nuclear area in cells with nuclear enlargement/nuclear area in normal intermediate cells) and nuclear contour thickening. High-risk HPV was detected using the Uniplex E6/E7 polymerase chain reaction assay and logistic regression analyses of factors related to high-risk HPV infection were performed. RESULTS: Nuclear contour thickening was present in 57.7% (64/111 cells) of high-risk HPV positive cases and 21.8% (69/317 cells) of high-risk HPV negative cases. There was a statistically significant association (P = 0.01) between high-risk HPV infection and nuclear contour thickening. Nuclear contour thickening was approximately one-third higher in NILM cases than in ASC-US cases (odds ratio, 0.371; 95% confidence interval, 0.208-0.662) and three times higher in high-risk HPV-positive cases than in high-risk HPV-negative cases (odds ratio, 2.831; 95% confidence interval, 1.591-5.039). CONCLUSION: Our results suggest that nuclear contour thickening in nuclear enlarged cells in NILM and ASC-US cases may be a cellular finding associated with HPV infection.

Significance of Compression in Binucleation while Differentiating Reactive Cellular Changes Between Human Papillomavirus and Candida Infections

Purpose: Binucleation is a reactive cellular change (RCC) in Pap smears due to Candida infection. However, the origin of these binucleated cells as RCCs remains unclear. The purpose of this study was to examine binucleation in patients negative for intraepithelial lesion or malignancy (NILM) and infected with Candida and those infected with high-risk human papillomavirus (hr-HPV) and to clarify the origin of the binucleated cells. Methods: A total of 115 endocervical swab specimens with a combined diagnosis of NILM, Candida infection, and RCCs were used for this study. Pap smears were used to identify binucleated cells and then separate them into two groups, compression-positive and compression-negative. In addition, hr-HPV was detected using polymerase chain reaction (PCR) with a specific primer on the DNA extracted from the remaining residual cytology specimens. To make the hr-HPV-infected binucleated cells visible, an in situ PCR assay was performed on the Pap smear. Result: Of the 115 specimens, 69.6% contained binucleated cells, 26 (32.5%) showed only the compressed form, 35 (43.8%) showed only the non-compressed form, and 19 showed both the compressed and non-compressed forms of binucleated cells. Also, 34 specimens (29.6%) were positive for hr-HPV. The sensitivity and specificity of compression-positive binucleated cells were 91.2% and 82.7% (p < 0.001), but they were not significant in the compression-negative group (p = 0.156). Also, 34 cases with hr-HPV contained 99 compression-positive and 24 compression-negative cells. The hr-HPV-positive cells accounted for 68 (68.7%) of the 99 compression-positive and 2 (8.3%) of the 24 compression-negative binucleated cells as determined by an in situ PCR assay for hr-HPV. The relationship between compression and hr-HPV was statistically significant (p < 0.001). Conclusion: Compression-positive binucleated cells may be present as a result of hr-HPV infection and not RCC, which is caused due to inflammation in NILM cases infected with Candida.

Cytological Features Associated with Ureaplasma Urealyticum in Pap Cervical Smear

Purpose: Ureaplasma urealyticum is associated with several obstetric complications and increases the importance of risk management in pregnant women. Furthermore, U. urealyticum has been identified as a cofactor that interacts with human papillomavirus infection in cervical cancer onset. The aim of this study was to assess specific cytological features of U. urealyticum infection in Pap smears to determine whether additional microbiological testing should be performed for pregnant women with a high possibility of U. urealyticum infection. Methods: Liquid-based cytology specimens (LBC) from cervical swabs of a total of 55 women, including 33 pregnant women who were negative for intraepithelial lesion or malignancy (NILM) on Pap testing and with U. urealyticum diagnosed without any other infectious microbes and 22 U. urealyticum-negative controls, were used in this study. We evaluated the localization of U. urealyticum by immunofluorescence, cytological features of secondary changes in squamous cells caused by inflammation, and the specimen background in Pap smears. Results: Based on analysis of Pap smears, a significant relationship was observed between U. urealyticum infection and cannonballs (p < 0.05) as well as predominance of coccoid bacteria (p < 0.05). A large number of U. urealyticum were detected in cannonballs by immunofluorescence. Conclusion: In the present study, cytological features in Pap smears of U. urealyticum infected samples, which have hardly been understood thus far, were assessed. The cytological features included cannonballs and predominance of coccoid bacteria. Our results might help in determining whether additional microbiological testing should be performed for pregnant women with a high possibility of U. urealyticum infection.

Evaluating the Existence of Small Compressed Binucleated Squamous Cells in ASC-H

Purpose: To evaluate the legitimacy of a diagnosis of ASC-H in 5 cases which were followed up monthly for over 2 years with both cytology and HPV testing. Methods: Some 5 cases out of a total of 25.0 self-sampled Pap test patients diagnosed as ASC-H provided 119 specimens over 2 years, with HPV-DNA testing perormed using a E6 primer. Results: Cases 1, 2 and 3 showed SIL after the ASC-H diagnosis, while cases 4 and 5 showed and maintained NILM. Cases 1, 2 and 3 were further characterized by small atypical compressed binucleated cells, in which HPV was detected by in situ PCR. Case 4 showed a high N/C ratio in cells in sheets with a mild increase in chromatin. Case 5 demonstrated a high N/C ratio in small cells with no increase in chromatin. Conclusion: The finding of a compressed binucleated cells can define the difference between degenerated endocervical columnar cells and small atypical cells suggestive of HSIL. When small compres

Improved accuracy of cytodiagnosis using the Kato self-collection devise: the usefulness of smear preparation in liquid-based cytology methods.

OBJECT: In the present study, we compared the positive cytodiagnostic test rates with discrepancies using self-collection devices for cervical cancer screening. We made this survey to examine whether or not our self- smear preparation method using the Kato self-collection device contributed to an improved rate of detecting atypical cells compared with existing recommended preparation methods. METHODS: Specimens were collected at 14 facilities handling self-collection methods, and samples were collected by a physician in 2 facilities. The chi- squared test was performed using the SPSS ver. 20 statistical software to determine the relationships between the positive cytodiagnostic rate, specimen preparation methods, and self-collection devices. RESULTS: Collecting cells using the Kato self-collection device and preparing liquid-based specimens, we obtained a significantly higher rate of positive cytodiagnosis and our results were equal to those obtained with the direct method. CONCLUSIONS: Taking into consideration increased needs for screening using the self-collection method in future, with even more improved test accuracy, a screening test that is acceptable to society needs to be established.