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.

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.

Standardization of fine needle aspiration cytology of the breast -- comparison of Auto Cyto Fix and conventional smears.

In Japan, there are some problems with fine needle aspiration (FNA) cytology of the breast, such as insufficient smeared cells, air-drying artefact and excessive erythrocytes. Liquid-based cytology has been found to solve these problems. Equipment for such preparations has been developed, but can be expensive to purchase and operate. We developed Auto Cyto Fix 1000 (ACF), which is inexpensive and automatically smears and fixes cells. The purpose of this study was to compare the various cytological features of conventional and ACF specimens. We evaluated whether the ACF method would be able to replace the conventional method. Forty-eight FNA specimens of breast were studied. All specimens were prepared by the direct smeared (DS) and ACF methods and evaluated for unsatisfactory cell collection, air-drying artefacts, background findings and epithelial cell findings. Although ACF specimens were prepared using the cells remaining in the needle and syringe after preparing DS specimens, the cellularity of two of the ACF specimens was better than that of the corresponding DS specimens. ACF specimens never showed air-drying artefact. Unlike DS specimens, which have many erythrocytes in the background, erythrocytes were filtered out and the background of ACF specimens was clean. We believe that many problems attributable to conventional FNA specimen preparation have been solved in this study. Preparation using the ACF apparatus can reduce running costs and can be used to prepare FNA specimens of the breast for cytological examination as an alternative to the conventional method.

Identification of "papillary-like" clusters on endometrial cytologic preparations.

The presence of papillary cell clusters is taken as a serious indication in the diagnosis of well-differentiated endometrial adenocarcinoma in Japan. However, papillary-like clusters have been observed in both normal and benign samples. Therefore, overdiagnosis may occur in the observation of cytopreparations. The present study attempts to prepare a histological sample from an Auto Cyto Fix (ACF) sample prepared by the automatic fixation apparatus (ACF1000) using a membrane filter method after cytological observation and to examine its usefulness. In an ACF sample, a papillary-like cluster was observed and the coverslip was then removed, the circumference of the membrane filter was cut off, and the target cell cluster was smeared, embedded, and sliced. In the cases in which a papillary-like cluster was observed, even if differential diagnosis of the derivation was difficult due to a resemblance between the respective morphological findings, it was easily made by histological observation of an ACF sample.

Usefulness of Auto Cyto Fix (membrane filter method) for the application of immunohistochemistry.

We developed the Auto Cyto Fix (ACF) 1000 as an experimental model, which was adapted as a membrane filter method for automatically making smear preparations fixed with 95% ethanol. In this study, immunohistochemistry was applied to the cells smeared on the membrane filter, and effectiveness was examined. Twenty-four effusions and HeLa 229 cells were used. These cell samples were produced by the ACF1000 on the membrane filter and stained by the Papanicolaou method. After observation, these were decolored and stained by immunohistochemical techniques. Antibodies included CEA, EMA, and MIB-1. CEA and EMA were detected by the indirect method, and were colored by DAB, AEC, and new fuchsin. MIB-1 was detected by the immunoperoxidase (LSAB) and the immunofluorescence method. A uniformly stabilized immunoreaction was obtained and was equivalent to or better than that of the conventional technique. In addition, there was no background staining on the membrane filter. Based on these results, the membrane filter preparation produced by the ACF1000 was also effectively applied to immunohistochemical observations.

Decreased nm23 expression, but not Ki-67 labeling index, is significantly correlated with lymph node metastasis of breast invasive ductal carcinoma.

The nm23 gene was originally identified by differential hybridization of metastatic murine melanoma cell lines. Some experimental studies demonstrated a significantly reduced metastatic potential of melanoma cell lines transfected with the nm23 gene. In this study, we clarified the relationship between lymph node status and nm23 immunoreactivity, as well as Ki-67 labeling index (LI), of human breast cancer. Of the 44 breast invasive ductal carcinomas, nm23-diffusely positive expression [nm23(+)] was detected in 17 (38.6%), and focally positive/negative expression [nm23(+/-/-)] in 27 (61.4%) cases. Lymph node metastasis was found at a significantly higher incidence in the nm23(+/-/-) cases (18/27, 66.7%) than in the nm23(+) cases (4/17, 23.5%) (p>0.001). In the lymph node metastasis-positive cases, mean LI of Ki-67 cells was 20.9% at the center of the tumors and 24.0% at the advanced margins. In the lymph node metastasis-negative cases, mean LI of Ki-67 cells was 12.4% at the center of the tumors and 27.2% at the advanced margins. Decrease of nm23 expression, but not Ki-67 LI, was significantly correlated with lymph node metastasis of breast invasive ductal carcinoma.