Molecular Signatures of High-Grade Cervical Lesions.

Cervical cancer is the fourth most common neoplasia in women and the infection with human papilloma virus (HPV) is its necessary cause. Screening methods, currently based on cytology and HPV DNA tests, display low specificity/sensitivity, reducing the efficacy of cervical cancer screening programs. Herein, molecular signatures of cervical cytologic specimens revealed by liquid chromatography-mass spectrometry (LC-MS), were tested in their ability to provide a metabolomic screening for cervical cancer. These molecules were tested whether they could clinically differentiate insignificant HPV infections from precancerous lesions. For that, high-grade squamous intraepithelial lesions (HSIL)-related metabolites were compared to those of no cervical lesions in women with and without HPV infection. Samples were collected from women diagnosed with normal cervix (N = 40) and from those detected with HSIL from cytology and colposcopy (N = 40). Liquid-based cytology diagnosis, DNA HPV-detection test, and LC-MS analysis were carried out for all the samples. The same sample, in a customized collection medium, could be used for all the diagnostic techniques employed here. The metabolomic profile of cervical cancer provided by LC-MS was found to indicate unique molecular signatures for HSIL, being two ceramides and a sphingosine metabolite. These molecules occurred independently of women's HPV status and could be related to the pre-neoplastic phenotype. Statistical models based on such findings could correctly discriminate and classify HSIL and no cervical lesion women. The results showcase the potential of LC-MS as an emerging technology for clinical use in cervical cancer screening, although further validation with a larger sample set is still necessary.

Interlaboratory quality control in gynecologic cytopathology using the novel CONQUISTADOR software. Interobserver reproducibility in the Latin American screening study.

OBJECTIVE: The recently developed software (CONQUISTADOR), capable of computing all intralaboratory and interlaboratory quality control (QC) indicators, was used to evaluate the diagnostic agreement among 4 cytology laboratories participating in the LAMS Study. STUDY DESIGN: The study was an interlaboratory exchange of specially designed 5 slide sets, each comprising 20 (conventional cytology) slides. At the first step, 80 slides (with "clear-cut" cases) were divided into four sets (A, B, C, D) of 20 specimens, each including inadequate and negative cases as well as in different proportions of all diagnostic TBS 2001 categories. In the second round, a fifth set (E) of 20 slides ("difficult cases") was designed, with all diagnostic categories, ASC and AGC included. Common measures of reproducibility (kappa and weighted kappa), accuracy (SE, SP, PPV, NPV) and 3 indices of diagnostic variability were calculated for sets A-D and set E, separately. RESULTS: For the 5 slide sets together, the weighted kappa was 0.8 (95% CI 0.76-0.85), which is the lower limit of the "almost perfect" ranking of kappa statistics, indicating an excellent interlaboratory agreement. The interlaboratory reproducibility was lower only for the difficult set (E). Similarly, the sensitivity for set E (70.0%) was lower than that (92.1%) for sets A-D. The diagnostic variability indices were not substantially different between the difficult (set E) and clearcut (sets A-D) cases. CONCLUSION: High interlaboratory reproducibility was obtained for sets A-D ("clear-cut" cases), while more interlaboratory variation was evident in the difficult samples. The new CONQUISTADOR software is a valuable tool in calculating the indicators needed in this intralaboratory and interlaboratory.

Fatores associados a resultados falso-negativos de exames citopatológicos do colo uterino / Factors associated with false-negative cervical cytopathological results

OBJETIVO: verificar se, no escrutínio de rotina, fatores relacionados com a adequabilidade da amostra, padrão celular e critérios citomorfológicos estão associados a resultados falso-negativos (FN) dos exames citopatológicos. MÉTODOS: trata-se de um estudo caso-controle, no qual o grupo de casos incluiu 100 esfregaços citopatológicos com um resultado FN que foi detectado pela sistemática de controle interno da qualidade com revisão rápida de 100 por cento. Para cada resultado FN detectado foram identificados, pelo mesmo citotécnico, dois esfregaços com um diagnóstico verdadeiro-positivo e este grupo foi considerado controle, totalizando uma casuística de 300 esfregaços. As variáveis analisadas foram estabelecidas de acordo com os critérios definidos para a análise da adequabilidade da amostra, padrão celular e critérios citomorfológicos. Os resultados foram avaliados por análise bivariada e regressão logística com critério de seleção de variáveis stepwise e expressos em OR (95 por cento). RESULTADOS: o número de células atípicas, aspecto da cromatina nuclear, distribuição e apresentação de células atípicas no esfregaço apresentaram risco maior para resultados FN, com OR de 9,6; 4,2; 4,4 e 3,6, respectivamente. Processo inflamatório e presença de sangue no esfregaço mostraram também risco para os resultados FN. CONCLUSÕES: a maioria dos fatores associados à liberação de um resultado FN é dependente das condições e técnicas de coleta de material, pois, em grande parte, a lesão pode não estar adequadamente representada no esfregaço, e também fatores obscurecedores como sangue e processo inflamatório podem prejudicar a análise. Quanto às alterações citomorfológicas, cromatina fina foi a característica que apresentou maior risco para resultados FN.

PURPOSE: to evaluate whether factors related to the adequacy of the sample, cell pattern and cytomorphological criteria are associated with false-negative (FN) results of cervical cytopathology during routine examinations. METHODS: this is a case-control study in which the study group included 100 cytopathologic smears with FN results detected during systematic internal quality control consisting of 100 percent rapid review. For each FN result detected, two smears with a true-positive diagnosis were identified by the same cytotechnician and these constituted the control group, making a total sample size of 300 smears. The variables were established in accordance with the criteria defined for the analysis of sample adequacy, cell pattern and cytomorphological analyzed criteria. The results were evaluated using bivariate analysis and logistic regression with stepwise variable selection criteria expressed in OR (95 percent). RESULTS: the number of atypical cells, the appearance of nuclear chromatin, and the distribution and presentation of atypical cells in the smear were the variables that showed the greatest risk for FN results with OR of 9.6, 4.2, 4.4, and 3.6, respectively. Inflammatory processes and the presence of blood in the smear were also identified as variables that influence the risk of FN results. CONCLUSIONS: the majority of the factors associated with FN results are dependent on the conditions and techniques of sample collection, since in the majority of cases, the lesion may not be adequately represented in the smear. Confounding factors such as blood and inflammatory processes may also impair analysis. With respect to cytomorphological alterations, thin chromatin strand was the variable that indicated the greatest risk of FN results.