Novaprep(®) Vial Test is a suitable liquid-based cytology medium for high risk human papillomavirus testing by Hybrid Capture 2.

BACKGROUND: Liquid-based cytology (LBC) for cervical cancer screening presents the advantage that cytological and virological investigations can be undertaken from the same specimen. Nevertheless, the fixative may alter DNA integrity and the sample may be inadequate for HPV DNA detection. The Novaprep(®) Vial Test (NVT) (Novacyt, Vélizy-Villacoublay, France) is a new device dedicated to LBC which permits an automated cell spreading over slides and an automated cell sampling for molecular analyses. OBJECTIVE: To determine whether the NVT was suitable for high risk (HR) HPV DNA detection with the Hybrid Capture 2 (HC2) assay (Qiagen, Courtaboeuf, France). STUDY DESIGN: Two cervical specimens were harvested. The first sample was taken with a Rovers Cervex Brush (Therapak Corporation, Buford, USA) placed in the NVT and the second sample was taken with a DNAPAP cervical sampler placed in the Specimen Transport Medium (STM) (Qiagen). This last sample served as gold standard for HPV detection. NVT and STM samples were analyzed for HR HPV DNA with HC2 assay. RESULTS: One hundred and thirty-one samples stored in NVT and STM were analyzed. The overall HC2 positivity determined from the 99 samples classified as satisfactory for cellularity (>5000 cells/slide) was 84% whatever the collection medium was. Agreement for HPV detection between NVT and STM was 94%, with a Kappa of 0.78. Moreover, we noted that HC2 values obtained from NVT samples were correlated to those obtained from STM samples. CONCLUSION: The Novaprep(®) Vial Test adequately preserves HPV DNA and is suitable for HPV testing with HC2 if cellularity is satisfactory.

[Progress report on the Bethesda System]. / Le point sur le Système de Bethesda 2001.

The 2001 Bethesda System is a uniform system of terminology for reporting results of pap smears. It is acknowledged by most cytopathologists worldwide as a standard for cervical cytology reports. In France, several national surveys have confirmed its current utilization. However, more specific analysis have shown that the Bethesda System may be routinely modified by individual laboratories or even individual cytopathologist working within the same department. The aim of this progress report was to emphasize the importance of fully understanding the Bethesda System and applying it in a rigorous and standardized way.

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.

Clinical and biological characteristics of cervical neoplasias with FGFR3 mutation.

BACKGROUND: We have previously reported activating mutations of the gene coding for the fibroblast growth factor receptor 3 (FGFR3) in invasive cervical carcinoma. To further analyze the role of FGFR3 in cervical tumor progression, we extended our study to screen a total of 75 invasive tumors and 80 cervical intraepithelial neoplasias (40 low-grade and 40 high-grade lesions). RESULTS: Using single strand conformation polymorphism (SSCP) followed by DNA sequencing, we found FGFR3 mutation (S249C in all cases) in 5% of invasive cervical carcinomas and no mutation in intraepithelial lesions. These results suggest that, unlike in bladder carcinoma, FGFR3 mutation does not or rarely occur in non invasive lesions. Compared to patients with wildtype FGFR3 tumor, patients with S249C FGFR3 mutated tumors were older (mean age 64 vs. 49.4 years, P = 0.02), and were more likely to be associated with a non-16/18 HPV type in their tumor. Gene expression analysis demonstrated that FGFR3 mutated tumors were associated with higher FGFR3b mRNA expression levels compared to wildtype FGFR3 tumors. Supervised analysis of Affymetrix expression data identified a significant number of genes specifically differentially expressed in tumors with respect to FGFR3 mutation status. CONCLUSION: This study suggest that tumors with FGFR3 mutation appear to have distinctive clinical and biological characteristics that may help in defining a population of patients for FGFR3 mutation screening.

Regression of low-grade cervical intraepithelial neoplasia in patients with HLA-DRB1*13 genotype.

OBJECTIVE: The human leukocyte antigen (HLA)-DRB1*13 allele frequency is lower in women with cervical carcinoma than in the general population, suggesting that this allele could exert a protective effect against progression of cervical intraepithelial neoplasia (CIN) associated with human papillomaviruses (HPV). To test this hypothesis, we designed a prospective study of low-grade CIN (CIN1) and analyzed the probability of regression of these lesions according to HLA-DR and HPV status. METHODS: The study sample was composed of 86 women with CIN1 who agreed to regular colposcopic follow-up and no immediate treatment. Biopsy specimens were taken under colposcopy for histology and for the determination of HPV and HLA status. Cases were classified into 3 groups: CIN1 regression, persistence for at least 12 months, or progression to CIN2 or 3. RESULTS: The rate of spontaneous regression (95% confidence interval) at 24 months was 51.6% (39-61.6%) overall compared with 34.7% (13.4-50.8%) in HPV16/18 positive cases and 59.9% (43.7-71.4%) in HPV16/18-negative cases (P =.051). The rate of regression was 71.8% (40.8-86.5%) in patients with HLA-DRB1*13 and 45.9% (31.5-57.2%) in patients with other genotypes (P =.03). Regression reached 90.5% (38.9-98.5%) at 18 months in DRB1*13 patients with HPV16/18-negative-associated CIN (15.1% of the cases). In multivariable analysis, HLA-DRB1*13 allele and HPV16/18-negative status were independently associated with an increased probability for regression (adjusted hazard ratio 2.1 [1.0-4.1] and 2.5 [1.2-5.4], respectively). CONCLUSION: A subset of approximately 15% of CIN1 highly likely to show spontaneous regression can be defined using 2 biologic parameters that characterize the viral causative agent and the host. LEVEL OF EVIDENCE: II-2

Efficiency of the hybrid capture 2 HPV DNA test in cervical cancer screening. A study by the French Society of Clinical Cytology.

The aim of this study was to determine the efficiency of the Hybrid Capture 2 (HC2; Digene, Gaithersburg, MD) human papillomavirus (HPV) assay for the detection of cervical neoplasia. Of the 1,785 patients recruited, 462 (25.88%) were referred for colposcopy owing to previously detected cytologic abnormalities, and 1,323 (74.12%) were voluntary candidates for screening. For all patients, a Papanicolaou smear and a monolayer smear (ThinPrep, Cytyc, Boxborough, MA) were done. HPV DNA was detected on the residual liquid-based material. False-positive results were observed in 111 cases and comprised 34 cross-reactions (1.90%) and 77 false-positive cases (4.31%) owing to a contiguous strong chemiluminescence signal. Interestingly, all these samples had a relative light units value of 1 to 3 and were contiguous to a sample with a very high HPV DNA load. The final results showed that high-risk and low-risk HPV DNA were detected in 480 samples (26.89%) and 135 samples (7.56%), respectively. Although HC2 can be considered a reliable and sensitive test for HPV DNA detection, we do not advocate its use for large-scale screening for cervical neoplasia.

Cross sectional study of conventional cervical smear, monolayer cytology, and human papillomavirus DNA testing for cervical cancer screening.

OBJECTIVES: To compare the sensitivity, specificity, and interobserver reliability of conventional cervical smear tests, monolayer cytology, and human papillomavirus testing for screening for cervical cancer. DESIGN: Cross sectional study in which the three techniques were performed simultaneously with a reference standard (colposcopy and histology). SETTING: Public university and private practices in France, with complete independence from the suppliers. PARTICIPANTS: 828 women referred for colposcopy because of previously detected cytological abnormalities and 1757 women attending for routine smears. MAIN OUTCOME MEASURES: Clinical readings and optimised interpretation (two blind readings followed, if necessary, by consensus). Sensitivity, specificity, and weighted kappa computed for various thresholds of abnormalities. RESULTS: Conventional cervical smear tests were more often satisfactory (91% v 87%) according to the Bethesda system, more reliable (weighted kappa 0.70 v 0.57), and had consistently better sensitivity and specificity than monolayer cytology. These findings applied to clinical readings and optimised interpretations, low and high grade lesions, and populations with low and high incidence of abnormalities. Human papillomavirus testing associated with monolayer cytology, whether systematic or for atypical cells of undetermined significance, performed no better than conventional smear tests. CONCLUSIONS: Monolayer cytology is less reliable and more likely to give false positive and false negative results than conventional cervical smear tests for screening for cervical cancer.

Cost of screening for cancerous and precancerous lesions of the cervix.

This paper is part of the cost-effectiveness study of cervical cancer screening conducted by the French Society of Clinical Cytology (SFCC). It describes the evaluation of costs of conventional smear tests, thin-layer smear tests (ThinPrep 2000 system), and viral typing by the HCS test. For 100,000 examinations per year, the average cost of a conventional smear test is 11.53 dollars in a private anatomo-pathology clinic. The cost of the thin-layer test for the same number of examinations and in the same type of clinic is 13.93 dollars. For 20,000 annual tests, the average cost of human papillomavirus (HPV) is 23.43 dollars in the public sector and 23.48 dollars in the private one. The higher price of the thin-layer method is only justifiable if this screening technique outperforms the conventional method. Furthermore, the high cost of the HPV test means that its integration into a population-based screening program must be carefully defined.