Comparison of Classical and Secondary Cytologic Criteria Relative to Hybrid Capture for Diagnosing Cervical-vaginal Infection by Human Papillomavirus

Objective To compare the diagnostic accuracy of the classic Meisels cytologic criteria and the Schneider secondary criteria relative to the hybrid capture method for diagnosing HPV infection. Methods This was a retrospective study performed at a public university hospital. A total of 41 patients with a cytologic diagnosis of HPV infection and 40 HPV-negative patients were selected for review of the cervical-vaginal smears seeking to classical and secondary criteria. A single pathologist reviewed the slides in search of the criteria. The classical and secondary cytologic criteria were compared with the hybrid capture for diagnosing HPV infection. Bartleti test was applied for the age analysis, and Fisher's exact test was used to compare proportions. The tests were considered significant when the probability of rejecting the null hypothesis was less than 5% (p < 0.05). Results The Meisels criteria were less sensitive (34.0%) than the secondary Schneider criteria (57.5%) when compared with the hybrid capture (p < 0.0001), although the specificity of the former criteria was non-significantly higher (91.2% and 67.7%, respectively). In cases of moderate or intense inflammation, the sensitivity and specificity of the Schneider criteria were decreased, 33.3% and 50.0% respectively (p = 0.0115). Conclusions Compared with hybrid capture for diagnosis of HPV infection, the sensitivity of the secondary Schneider criteria was higher than the classical Meisels criteria.Moderate or intense inflammation reduces the sensitivity and specificity of the secondary Schneider criteria for diagnosing HPV infection using the hybrid capture as the gold standard.

Factors related to inadequate cervical cancer screening in two Brazilian state capitals

OBJECTIVE:To analyze factors associated with cervical cancer screening failure. METHODS:Population-based cross-sectional study with self-weighted two-stage cluster sampling conducted in the cities of Fortaleza (Northeastern Brazil) and Rio de Janeiro (Southeastern Brazil) in 2002. Subjects were women aged 25-59 years in the last three years prior to the study. Data were analyzed through Poisson regression using a hierarchical model. RESULTS: The proportion of women who did not undergo the Pap smear test in Fortaleza and Rio de Janeiro was 19.1% (95% CI: 16.1;22.1) and 16.5% (95% CI: 14.1;18.9), respectively. Higher prevalence ratios of cervical cancer screening failure in both cities were seen among women with low education and low per capita income, old age, unmarried, who never underwent mammography, clinical breast examination, and blood glucose and cholesterol level testing. Smokers also had lower screening rates compared to non-smoker women and this difference was only statistically significant in Rio de Janeiro. CONCLUSIONS:The study findings point to the need of intervention focusing particularly women in worse socioeconomic conditions and access to healthcare, old-aged and unmarried. Education activities must prioritize screening of asymptomatic women and early diagnosis for symptomatic women and access to adequate diagnostic methods and treatment should be provided.

A morphological protocol and guide-list on uterine cervix cytology associated to Papillomavirus infection

The present study was designed to further assess the validity of the cytological description of morphological lesions said to be related to Papillomavirus (HPV) infections in senior women. The casuistic comprised 196 cervical smears from a group of women with no clinical or morphological evidence of neoplasia, collected simultaneously with samples submitted to detection of HPV DNA by PCR in a previous study. Three experienced cytologists studied each slide in two different conditions, with an interval of 20 months between them. The first approach was performed under routine laboratory standards, whereas the second was guided by a list of 16 well-defined parameters indicative of HPV-related cytological lesions. When suspicious cases of HPV-related alterations were grouped with positive cases, they showed on average: sensitivity of 25.5%, specificity of 84.4% and positive predictive value (PPV) of 26.8%. When suspicious cases were grouped with negative cases, sensitivity decreased, whereas specificity and PPV increased, as expected. In the second reading, which followed a "guide-list", a decrease in sensitivity was observed, contrasting with a sharp increase of positive predictive value. Among the 16 cytomorphological criteria tested, "koilocytosis", "mild koilocytosis" and "condylomatous parabasal cells" yielded the best predictive value for HPV DNA detection by PCR. In conclusion, despite the low sensitivity, cytopathologic assessment of cervico-vaginal smears leads to a highly specific diagnosis of HPV infection in menopausal women, with PPV of 91.0% when directed by a guide-list of well-defined morphologic criteria.

Flora vaginal e correlação com aspectos citológicos

Com a finalidade de esclarecer a patogenicidade de microorganismos que compõem a microflora vaginal, particularmente nas chamadas vaginites inespecíficas e de determinar o valor da citologia como método para identificação da flora vaginal, foram estudadas 97 pacientes, realizando cultura, exame direto a fresco, Gram e citologia do material cérvico-vaginal. Entre os microorganismos de importância clínica reconhecida, Gardnerella vaginalis foi o mais freqüentemente isolado, 48,4%, seguido de Trichomonas vaginalis, 10,3%, Candida albicans, 7,2% e Neisseria gonorrhoeae, 1,1%. Alterações citológicas indicativas de cervicite e/ou vaginite estiveram presentes na maioria dos casos de G.vaginalis, C.albicans e em todos os casos de T.vaginalis. Foi ressaltada a importância da avaliação semi-quantitativa dos microorganismos nos meios de cultura. Na avaliação da citologia como método diagnóstico para a microflora vaginal, foi observada que este foi o melhor dos métodos utilizados na identificação de T.vaginalis, tendo sido detectado cerca de 50% dos casos de C.albicans. No diagnóstico de G.vaginalis a citologia foi positiva em 48,9% dos casos, destacando-se que dos 31 casos positivos ao exame citológico, oito tiveram culturas negativas para G.vaginalis, embora tenham sido isolados nestes últimos: Haemophilus sp. ou Corynebacterium sp.

Risk factors for the increasing trend in low birth weight among live births born by vaginal delivery, Brazil

OBJECTIVE: To identify risk factors for low birth weight (LBW) among live births by vaginal delivery and to determine if the disappearance of the association between LBW and socioeconomic factors was due to confounding by cesarean section. METHODS: Data were obtained from two population-based cohorts of singleton live births in Ribeirão Preto, Southeastern Brazil. The first one comprised 4,698 newborns from June 1978 to May 1979 and the second included 1,399 infants born from May to August 1994. The risks for LBW were tested in a logistic model, including the interaction of the year of survey and all independent variables under analysis. RESULTS: The incidence of LBW among vaginal deliveries increased from 7.8% in 1978--79 to 10% in 1994. The risk was higher for: female or preterm infants; newborns of non-cohabiting mothers; newborns whose mothers had fewer prenatal visits or few years of education; first-born infants; and those who had smoking mothers. The interaction of the year of survey with gestational age indicated that the risk of LBW among preterm infants fell from 17.75 to 8.71 in 15 years. The mean birth weight decreased more significantly among newborns from qualified families, who also had the highest increase in preterm birth and non-cohabitation. CONCLUSIONS: LBW among vaginal deliveries increased mainly due to a rise in the proportion of preterm births and non-cohabiting mothers. The association between cesarean section and LBW tended to cover up socioeconomic differences in the likelihood of LBW. When vaginal deliveries were analyzed independently, these socioeconomic differences come up again.

Isolamento, identificação e sorotipagem de candida albicans a partir de secreção vaginal

Foram estudadas 104 amostras de secreção vaginal de mulheres com suspeita de candidiase segundo observações clinicas, na cidade de Alfenas-MG. Encontrou-se 55,7% de positividade para Candida albicans .prevalecendo maior índice na raça negra (64% de 25 amostras), sendo de 53,1% (79 amostras), a positividade na raça branca. Em 14 gestantes, a pesquisa da levedura mostrou-se positiva na totalidade dos casos. A maioria das amostras positivas (93,1%) procedia de mulheres com idade compreendida entre 20 40 anos. O uso de anticoncepcionais, antibióticos e presença de displasias cervicais mostraram-se como fatores que contribuiram para maior incidência do fungo. Das 58 amostras de C. albicans isoladas, 50 (86,2%) pertenciam ao sorotipo "A", sendo 37 (74%) isoladas de mulheres da raça branca e 13 (26%) da raça negra. Apenas 08 amostras (13,8%) pertenciam ao sorotipo "B", sendo 05 (11,9%) isoladas a partir de mulheres da raça branca e 03 (18,75%) da raça negra.

Identification of Cryptosporidium spp. oocysts in fecal smears stained with Heidenhain's iron hematoxylin

There is no paucity of methods for diagnosing Cryptosporidium spp. infection. The merits of immunoassays notwithstanding, microscopic identification of Cryptosporidium spp. oocysts in fecal samples remains an important diagnostic procedure. It owes the persistence of its use to such characteristics as dispensing with expensive equipment and kits, requiring only basic laboratory facilities, and having a low probability of false positive results when permanent slides are prepared, which can be re-examined in case of doubt. Cryptosporidium spp. oocysts can be readily identified in fecal smears prepared according to a regressive iron hematoxylin staining technique. The number of steps and their duration, as well as costs, were reduced to a minimum without loss of image quality and permanence of the preparations.

EFFECT OF HIGHLY ACTIVE ANTIRETROVIRAL THERAPY ON VAGINAL Candida spp. ISOLATION IN HIV-INFECTED COMPARED TO HIV-UNINFECTED WOMEN

 Vulvovaginal candidiasis (VVC) in HIV-infected women contributed to the impairment of their quality of life. The aim of this study was to evaluate the effect of highly active antiretroviral therapy (HAART) use on the vaginal Candida spp. isolation in HIV-infected compared to HIV-uninfected women. This cross-sectional study included 178 HIV-infected (HIV group) and 200 HIV-uninfected women (control) that were studied at the Specialized Assistance Service (SAE) for sexually transmitted diseases (STD)/AIDS of the city of Maringá, Brazil, from April 1 to October 30, 2011. The yeasts were isolated and identified by phenotypic and molecular methods. The in vitro antifungal susceptibility to fluconazole, itraconazole, nystatin and amphotericin B was tested by the reference microdilution method. Higher frequencies of total vaginal Candida spp. isolation were found in the HIV-infected group than in the control group. However, both groups showed a similar frequency of colonization and VVC. Although C. albicans was the most frequent and sensitive to azolics and polyenes in both HIV-infected and uninfected women, the emerging resistance of C. glabrata to amphotericin B in the HIV-infected women was observed. Although higher frequency of vaginal Candida spp. isolation had been observed in the HIV-infected than in HIV-uninfected women, colonization and VVC showed similar frequency in both groups, indicating that HAART appears to protect against vaginal colonization and VVC.

A key for identifying faecal smears to detect domestic infestations of triatomine bugs

Early detection of residual populations of domestic triatomine bugs that survive insecticide treatment is a key component of successful evaluation and vigilance for Chagas disease control. We have recently demonstrated that sheets of paper, tacked on to the walls of infested houses, can become streaked with the faeces of triatomine bugs and thus reveal thepresence of an infestation. In thispaper, wepresent a simple key to differentiate the faecal streaks of triatomine bugs from those of other domestic arthropods such as cockroaches, ticks and cimicid bedbugs.

Quadro citológico vaginal, concentração plasmática de progesterona durante a gestação e medidas fetais em paca (Cuniculus paca Linnaeus, 1766)

Em 27 pacas (Cuniculus paca Linnaeus, 1766) objetivou-se descrever aos 30, 60 e 90 dias (D) de prenhez diagnosticada por ultrassonografia (US), os tipos celulares do epitélio vaginal em esfregaços vaginais, relatar as condições de abertura da vulva e as características do muco vaginal, determinar a concentração plasmática de progesterona (P4) por radioimunoensaio, e ainda, mensurar por ultrassonografia (US) o diâmetro biparietal (DBP) fetal aos 60 e 90 dias de prenhez. No D30, 40% das amostras exibiram células (com características estrogênicas) superficiais e presença de núcleos nus. Nos D60 e D90, células parabasais, intermediárias, superficiais e naviculares estavam presentes nas mesmas proporções, mas células endocervicais foram descritas em apenas 73,9% e 69% das amostras daqueles dias, respectivamente. No D30 a maior proporção de células naviculares e superficiais diferiu (p < 0,05) em relação aos outros tipos celulares presentes. O muco vaginal apresentou-se cristalino e fluido em 100% e em 70% das fêmeas nos D30 e D60, respectivamente. Observou-se o vestíbulo vaginal aberto em torno de 50% das fêmeas em todos os dias de exames. Valores mínimos detectáveis de P4 foram obtidos em 72% e em 83% das fêmeas, enquanto que as médias das medidas dos DBP foram 1,25 cm (± 0,16) e 2,34 cm (± 0,25) nos D60 e D90, respectivamente. O quadro citológico vaginal nos D30, D60 e D90 e o DBP fornecem elementos que contribuem para diagnóstico de gestação em pacas. A concentração de P4 demonstra a necessidade de maiores estudos da endocrinologia da gestação em pacas.

Survival of tubercle bacilli in heat-fixed and stained sputum smears

We used a slide culture technique to detect tubercle bacilli surviving in sputum smears (n=46) after conventional heat fixation and Ziehl-Neelsen staining. In all heat-fixed sputum smears, tubercle bacilli survived after time 0 (n=22), 24 h (n=7), 48 h (n=7), 72 h (n=4), and seven days (n=6). None of the stained sputum smears showed growth on slide cultures. Viable tubercle bacilli remaining in heat-fixed sputum smears for at least seven days may present an infection risk to laboratory staff. Thus, sputum smears should be stained immediately by the Ziehl-Neelsen method or stored in a safe container to avoid transmission of tuberculosis.

First characterization of Candida albicans by random amplified polymorphic DNA method in Nicaragua and comparison of the diagnosis methods for vaginal candidiasis in Nicaraguan women

A total of 106 women with vaginitis in Nicaragua were studied. The positive rate for the identification of Candida species was 41% (44 positive cultures out of 106 women with vaginitis). The sensitivity of microscopic examination of wet mount with the potassium hydroxide (KOH) was 61% and 70% with Gram's stain when using the culture of vaginal fluid as gold standard for diagnosis of candidiasis. Among the 44 positives cultures, isolated species of yeast from vaginal swabs were C. albicans (59%), C. tropicalis (23%), C. glabrata (14%) and C. krusei (4%). This study reports the first characterization of 26 C. albicans stocks from Nicaragua by the random amplified polymorphic DNA method. The genetic analysis in this small C. albicans population showed the existence of linkage disequilibrium, which is consistent with the hypothesis that C. albicans undergoes a clonal propagation.

Identification of Mycobacterium tuberculosis complex based on amplification and sequencing of the oxyR pseudogene from stored Ziehl-Neelsen-stained sputum smears in Brazil

A cross-sectional analysis of stored Ziehl-Neelsen (ZN)-stained sputum smear slides (SSS) obtained from two public tuberculosis referral laboratories located in Juiz de Fora, Minas Gerais, was carried out to distinguish Mycobacterium bovis from other members of the Mycobacterium tuberculosis complex (MTC). A two-step approach was used to distinguish M. bovis from other members of MTC: (i) oxyR pseudogene amplification to detect MTC and, subsequently, (ii) allele-specific sequencing based on the polymorphism at position 285 of this gene. The oxyR pseudogene was successfully amplified in 100 of 177 (56.5%) SSS available from 99 individuals. No molecular profile of M. bovis was found. Multivariate analysis indicated that acid-fast bacilli (AFB) results and the source laboratory were associated (p < 0.05) with oxyR pseudogene amplification. SSS that were AFB++ SSS showed more oxyR pseudogene amplification than those with AFB0, possibly due to the amount of DNA. One of the two source laboratories presented a greater chance of oxyR pseudogene amplification, suggesting that differences in sputum conservation between laboratories could have influenced the preservation of DNA. This study provides evidence that stored ZN-SSS can be used for the molecular detection of MTC.