Immunocytochemical expression of p16 INK4A and Ki-67 in cytologically negative and equivocal pap smears positive for oncogenic human papillomavirus

This study was designed to analyze the cross-sectional comparison of the p16 INK4A and Ki-67 immunocytochemical expression in negative and equivocal (atyp-ical squamous cells of undetermined significance (ASC-US)) liquid-based cytology (LBC) samples testing positive for high-risk human papillomavirus (HPV) types with HC2 assay or polymerase-chain reaction (PCR). A series of 199 consecutive LBC speci-mens derived from the same number of women participating in the ongoing Latin American Screening Study at Leonor Mendes de Barros Hospital, São Paulo, were analyzed using immunocytochemistry for expression of p16 INK4A and Ki-67 in negative and equivocal LBC samples testing positive for high-risk HPV types with hybrid capture II test (HC2) or PCR. All patients with at least one test positive (cytology, PCR, and/or HC2) were followed each 6 months for 3 years. The follow-up procedure consisted of visual examination, colposcopic inspection, cytology, and HC2 assay. Among the neg-ative cytologic samples, 101 were HPV-positive and 55 HPV-negative. Of the HPV-pos-itive group, 59 of 101 cases (58.4%) were positive for both p16 and Ki67 immunostaining, and 17 of 101 (16.8%) were negative for both. The proportion of Ki-67-positivity increased almost in parallel with the increasing grade of p16-positivity (p = 0.0001 for linear trend). In the HPV-negative group, both markers were negative in 41 of 55 cases (74.5%), and no statistical relationship was observed between the two markers (Pearson, p = 0.595). HPV-positive ASC-US samples demonstrated a simulta-neous positive immunoreaction for p16 and Ki67 in 11 of 16 cases (68.7%), whereas 3 (18.7%) were concurrently negative. The relationship between the two markers was of borderline significance (Pearson, p = 0.053), but no linear relationship was found be-tween the graded p16 and Ki-67 expression (p = 0.065 for linear trend). In the HPV-negative ASC-US group, there was no statistical association between the graded p16 and Ki-67 positivity (Pearson, p = 0.281). After 36 months of follow-up of the ASC-US patients, 6 women still displayed ASC-US smear, of which 4 of 6 were HPV-positive and expressed both p16 and Ki-67 markers. Two of 43 ASC-US smears had high-grade squamous intraepithelial lesions diagnosed (4.6%), and 1 had low-grade squamous intraepithelial lesion (2.3%). All of those were positive for HPV, p16 and Ki-67. Patients with ASC-US diagnosis and positive high-risk HPV status and positive for p16 INK4A Ki67 should be carefully observed to exclude occurrence of a squamous intraepithelial lesion. The combination of these two markers can be a useful implement for manage-ment of women with equivocal cytology.European Commission (EC) - INCO-DEV Programme - Project #ICA4-CT-2001-10013

Squamous cervical intraepitelial bneoplasia grades II and III cytopathological and histopathological correlation: evaluation of the glandular involvement and of the cytophatic effects of human papillomavirus infection

O surgimento do Sistema de Bethesda para diagnosticos citologicos cervicovaginais trouxe varios questionamentos relativos a sua aplicabilidade em nosso meio, entre os quais: - agrupamento das alteracoes citologicas classificadas como NIC II e NIC III sob a denominacao unica de Lesao lntraepitelial Escamosa de Alto Grau; - utilizacao do diagnostico citologico de atipias em celulas glandulares de significado indeterminado (AGUS) na emissao de laudos citologicos. Outra questao amplamente discutida se refere a concordancia cito-histopatologica em relacao aos efeitos citopaticos da presenca de HPV nas Lesoes de Alto Grau. Como consequencia da utilizacao de criterios nao especificos para este diagnostico, o confronto de laudos incompativeis e bastante frequente em nosso meio. Na tentativa de elucidar estas questoes foram revistos 98 casos cuja citologia diagnosticou Lesao lntraepitellal Escamosa de Alto Grau e que foram submetidos a resseccao parcial do colo uterino atraves de cirurgia de alta frequencia, apos comprovacao diagnostica de NIC II ou NIC III por biopsia dirigida pela colposcopia. Os resultados mostraram que ha concordancia cito-histopatologica em 81,6 por cento dos casos, sendo maior nos casos de NIC III (87,1 por cento) do que nos de NIC II (67,9 por cento). Atraves da Analise de Regressao Logistica pudemos observar que a chance de se encontrar extensao glandular nas pacientes que apresentaram AGUS a citologia e 5,32 vezes maior em relacao aquelas que nao apresentaram e e 5,74 vezes maior nas lesoes NIC III do que nas NIC II. Os resultados tambem mostraram que a concordancia cito-histopatologica referente a presenca ou nao de efeitos citopaticos do HPV e de 60,7 por cento nos casos de NIC II, com valor preditivo positivo de 86,7 por cento e valor preditivo negativo de 30,8 por cento, e de 52,8 por cento nos casos de NIC III, com valor preditivo positivo de 72,2 por cento e valor preditivo negativo de 46,2 por cento, sempre utilizando coilocitos como criterio especifico para a presenca do efeito citopatico viral. O presente estudo pennite-nos concluir que e mais apropriado utilizar a nomenclatura sugerida pelo Sistema de Bethesda associada a nomenclatura ja estabelecida de NIC II e NIC III; o valor preditivo da citologia no comprometimento glandular das lesoes NIC II e NIC III e estatisticamente significante, quando utilizados criterios e objetivos para o diagnostico de AGUS; existe...(au)BV UNIFESP: Teses e dissertaçõe

Cervical biopsy-based comparison of a new liquid based TLP with conventional smears

Objetivo: Comparar a acuracia diagnostica de amostras citologicas em meio liquido UCM (Digene Corporation, MD, EUA) e de amostras citologicas convencionais utilizando-se o resultado histologico como padrao ouro. Material e Metodos: Estudo transversal em 788 mulheres submetidas a exame ginecologico durante o qual foram coletadas amostras cervicovaginais para a realizacao de citologia por ambos os metodos, seguidas de colposcopia e de biopsia se necessario. A avaliacao citologica pelos dois metodos foi separadamente feita. A concordancia entre os dois metodos citologicos foi verificada pela estatistica Kappa. Sensibilidade, especificidade e valores preditivos com intervalos de confianca de 95°lo foram calculados para ambos os metodos. O teste de McNemar foi empregado para determinar o nivel de coincidencia entre os dois procedimentos citologicos. Resultados: A concordancia entre os dois metodos em relacao ao diagnostico relatado mostrou um valor de Kappa de 0,774 (p < 0,0001). Apos ajuste para cada categoria de diagnostico histologico, o valor de Kappa encontrado para cada uma delas foi o seguinte: normal =0,733 (p < 0,0001); LSIL = 0,654 (p < 0,0001); HSIL = 0,735 (p < 0,0001). A sensibilidade e a especificidade da citologia UCM foram de 75,3 por cento e 86,2 por cento respectivamente, estatisticamente nao significante quando comparados aos 80,8 por cento e 85,0 por cento observados na citologia convencional. Conclusao: Este estudo demonstrou que nao houve diferenca entre a citologia em meio liquido UCM e o esfregaco convencional de Papanicolaou na deteccao de lesoes intra-epiteliais escamosas e do cancer do colo do uteroBV UNIFESP: Teses e dissertaçõe

Performance of 3 methods for quality control for gynecologic cytology diagnoses

Prova tipográfica (In Press)ObjectiveTo evaluate performance and viability of internal quality control QC) strategies in a public health laboratory of the state of Sao Paulo.Study DesignA retrospective study was per formed with 3 QC strategies to improve internal cytologic diagnoses: morphologic guided-list criteria (MGLC), 100% rapid-rescreening (100% RR) of negative slides ("turret" method) and 10% rescreening (10% R) of negative slides. Cases were examined at Adolfo Lutz Institute, Sao Paulo, Brazil, from 2002 to 2004. Histopathologic results, when available, were considered gold standard; cytologic consensus diagnosis was by 2 pathologists when histologic results were unavailable.ResultsMGLC selected 20.7% samples with cytologic atypias, 10% R selected 0.6% and RR selected 2.5%. Cytologic/histologic initial concordance was 57.4%, low-grade squamous intraepithelial lesion false negative rate was 34.9% and high-grade squamous intraepithelial lesion false negative rate was 12.2%. After diagnosis, consensus concordance was 97.2%.ConclusionThe 100% RR and 10% R QC strategies detected more false negative cases in liquid-based cytology than in conventional Pap smears. The 100% RR strategy reduced the false negative results and allowed evaluation of individual staff performance. The 10% R strategy did not offer significant results. We concluded that association of MGLC and 100% RR strategies might improve cytologic diagnostic quality

100% rapid rescreening for quality assurance in a quality control program in a public health cytologic laboratory

To verij5 the efficacy of the quality control (QC) program in a cytologic laboratwy with a rapid rescreening (RR) protocol. To verij5 the efficacy of the quality control (QC) program in a cytologic laboratwy with a rapid rescreening (RR) protocol.RR, according to the Turret RR method, of all samples initially screened as negative at the Laboratory of Cytology, Adolfo Lutz Institute, was performed. The slides were reviewed for 60 seconds. Suspect smears were fully checked by 2 reviewers to determine the final diagnoses. A total of 2954 sequential cytologic results were considered in this study. Of the 2954, 2568 (86.9%) were considered initially negative according to our internal QC, and these cases underwent RR. Also, 10% were randomly selected from these negative cases for full reviewing. The internal QC in our laboratory includes review of cases selected according to clinical and cytomorphologic criteria.Among the 2954 total cases, QC detected 386 (13%) atypias with final diagnoses reported according to The Bethesda System 2001 as follows: 82 (2.18%) low grade squamous intraepithelial lesions (LSILs), 35 (1.18%) high grade squamous intraepithelial lesions (HSILs), 2 (0.06%) squamous cell carcinomas, 105 (3.5%) atypical cells of undetermined significance (ASC-US), 4 (0.12%) atypical endocervical cells (AECs) and 158 (5.3%) unsatisfactory samples. RR of 2568 smears initially considered negative selected 194 (7.5%) slides. Of the 194, 146 (75.3%) were negative, 28 (14.4%) ASC-US, 5 (2.6%) AEC, 1 (0.5%) LSIL and 14 (7.2%) unsatisfactory. Full review of a 10% random fraction of the 2568 cases interpreted as negative did not detect lesions but did detect 5 (1.95%) unsatisfactory samples.Internal QC used in our laboratory based on clinical and cytomorphologic criteria to select cases for review proved to be an efficient method of detecting HSIL and cervical cancer. The consensus basis of this program strongly limits the false positive and false negative rates and also provides subjects with continuing education. One hundred percent RR is more efficient than 10% full reviewing in detecting cervical abnormalities