Estudo clínico com o uso do itraconazol em tinha crural, tinha do corpo e tinha do pé interdigital

Os autores estudaram 31 pacientes portadores de tinhas crural, do corpo e do pé interdigital, tratados com 100 mg de itraconazol, administrados junto com o almoço, durante 15 dias. Foram realizadas 3 avaliações clínicas e micológicas: no pré-tratamento, no final do tratamento e 2 semanas após o tratamento. O itraconazol mostrou-se eficaz para o tratamento das dermatofitoses estudadas, na dose empregada, com 100% de cura micológica. Em relação a evolução clínica, houve diminuição estatística significante (teste de Friedman - p < 0,001) dos sinais e sintomas avaliados. Não foi observada ocorrência de efeitos colaterais.

Optimization of the Sybr Green real time PCR for the detection of Human Herpes Virus type 6 (HHV-6)

HHV-6 is the etiological agent of Exanthem subitum which is considered the sixth most frequent disease in infancy. In immuno-compromised hosts, reactivation of latent HHV-6 infection may cause severe acute disease. We developed a Sybr Green Real Time PCR for HHV-6 and compared the results with nested conventional PCR. A 214 pb PCR derived fragment was cloned using pGEM-T easy from Promega system. Subsequently, serial dilutions were made in a pool of negative leucocytes from 10-6 ng/µL (equivalent to 2465.8 molecules/µL) to 10-9 (equivalent to 2.46 molecules/µL). Dilutions of the plasmid were amplified by Sybr Green Real Time PCR, using primers HHV3 (5' TTG TGC GGG TCC GTT CCC ATC ATA 3)'and HHV4 (5' TCG GGA TAG AAA AAC CTA ATC CCT 3') and by conventional nested PCR using primers HHV1 (outer): 5'CAA TGC TTT TCT AGC CGC CTC TTC 3'; HHV2 (outer): 5' ACA TCT ATA ATT TTA GAC GAT CCC 3'; HHV3 (inner) and HHV4 (inner) 3'. The detection threshold was determined by plasmid serial dilutions. Threshold for Sybr Green real time PCR was 24.6 molecules/µL and for the nested PCR was 2.46 molecules/µL. We chose the Real Time PCR for diagnosing and quantifying HHV-6 DNA from samples using the new Sybr Green chemistry due to its sensitivity and lower risk of contamination.

A real-time quantitative assay for hepatitis B DNA virus (HBV) developed to detect all HBV genotypes

Hepatitis B virus (HBV) is a major cause of chronic liver disease worldwide. Besides genotype, quantitative analysis of HBV infection is extensively used for monitoring disease progression and treatment. Affordable viral load monitoring is desirable in resource-limited settings and it has been already shown to be useful in developing countries for other viruses such as Hepatitis C virus (HCV) and HIV. In this paper, we describe the validation of a real-time PCR assay for HBV DNA quantification with TaqMan chemistry and MGB probes. Primers and probes were designed using an alignment of sequences from all HBV genotypes in order to equally amplify all of them. The assay is internally controlled and was standardized with an international HBV panel. Its efficacy was evaluated comparing the results with two other methods: Versant HBV DNA Assay 3.0 (bDNA, Siemens, NY, USA) and another real-time PCR from a reference laboratory. Intra-assay and inter-assay reproducibilities were determined and the mean of CV values obtained were 0.12 and 0.09, respectively. The assay was validated with a broad dynamic range and is efficient for amplifying all HBV genotypes, providing a good option to quantify HBV DNA as a routine procedure, with a cheap and reliable protocol.

MULTIPLEX SYBR® GREEN-REAL TIME PCR (qPCR) ASSAY FOR THE DETECTION AND DIFFERENTIATION OF Bartonella henselae AND Bartonella clarridgeiae IN CATS

A novel SYBR® green-real time polymerase chain reaction (qPCR) was developed to detect two Bartonellaspecies, B. henselae and B. clarridgeiae, directly from blood samples. The test was used in blood samples obtained from cats living in animal shelters in Southern Brazil. Results were compared with those obtained by conventional PCR targeting Bartonella spp. Among the 47 samples analyzed, eight were positive using the conventional PCR and 12 were positive using qPCR. Importantly, the new qPCR detected the presence of both B. henselae and B. clarridgeiae in two samples. The results show that the qPCR described here may be a reliable tool for the screening and differentiation of two important Bartonella species.

QUANTITATIVE REAL-TIME PCR (Q-PCR) FOR SPUTUM SMEAR DIAGNOSIS OF PULMONARY TUBERCULOSIS AMONG PEOPLE WITH HIV/AIDS

Objective: To assess quantitative real-time polymerase chain reaction (q-PCR) for the sputum smear diagnosis of pulmonary tuberculosis (PTB) in patients living with HIV/AIDS with a clinical suspicion of PTB.Method: This is a prospective study to assess the accuracy of a diagnostic test, conducted on 140 sputum specimens from 140 patients living with HIV/AIDS with a clinical suspicion of PTB, attended at two referral hospitals for people living with HIV/AIDS in the city of Recife, Pernambuco, Brazil. A Löwenstein-Jensen medium culture and 7H9 broth were used as gold standard.Results: Of the 140 sputum samples, 47 (33.6%) were positive with the gold standard. q-PCR was positive in 42 (30%) of the 140 patients. Only one (0.71%) did not correspond to the culture. The sensitivity, specificity and accuracy of the q-PCR were 87.2%, 98.9% and 95% respectively. In 39 (93%) of the 42 q-PCR positive cases, the CT (threshold cycle) was equal to or less than 37.Conclusion: q-PCR performed on sputum smears from patients living with HIV/AIDS demonstrated satisfactory sensitivity, specificity and accuracy, and may therefore be recommended as a method for diagnosing PTB.

Comportamento do peso do coração e do corpo em chagásicos crônicos com e sem "megas"

O peso do coração é menor nos chagásicos com "megas" do que nos falecidos subitamente ou em I.C.C., sendo, porém, maior que nos controles. Já o peso corporal ésemelhante nos chagásicos falecidos subitamente e nos controles, havendo redução moderada no grupo com I.C.C., e rtiais acentuada nos chagásicos com "megas" (redução de 20% em relação aos controles). A relação percentual peso cardíaco/peso corporal é de 1,1% no grupo com I.C.C.; 0,71% no grupo de morte súbita; 0,77% no grupo com "megas"; 0,49% no grupo controle e 0,60% ne grupo de doenças caquetizantes. Com base nas médias dos pesos cardíacos e corporal dos grupos controle, de doenças caquetizantes e de megas, calculou-se que o emagrecimento ocorrido nos megas reduziria o peso cardíaco médio de apenas 6,8%.

Carga proviral do HTLV-1 e HTLV-2: um método simples através da PCR quantitativa em tempo real

Os vírus linfotrópicos de células T humanas, quando integrados ao genoma da célula hospedeira, provírus, têm como marcador de replicação seu DNA proviral. A carga proviral parece ser um importante fator no desenvolvimento de patologias associadas a estes retrovírus. Neste estudo foi desenvolvida uma metodologia para quantificação absoluta da carga proviral dos HTLV-1 e HTLV-2 através da PCR em tempo real. Cinqüenta e três amostras de doadores de sangue com teste de ELISA reagente foram submetidas à metodologia, que utilizou o sistema TaqMan® para três seqüências alvo: HTLV-1, HTLV-2 e albumina. A quantificação proviral absoluta foi determinada através da proporção relativa entre o genoma do HTLV e o genoma da célula hospedeira, levando em consideração o número de leucócitos. O método apresentado é sensível (215 cópias/mL), prático e simples para quantificação proviral, além de eficiente e adequado para confirmação e discriminação da infecção pelos tipos virais.

Evaluation of the use of real-time PCR for human T cell lymphotropic virus 1 and 2 as a confirmatory test in screening for blood donors

INTRODUCTION: HTLV-1/2 screening among blood donors commonly utilizes an enzyme-linked immunosorbent assay (EIA), followed by a confirmatory method such as Western blot (WB) if the EIA is positive. However, this algorithm yields a high rate of inconclusive results, and is expensive. METHODS: Two qualitative real-time PCR assays were developed to detect HTLV-1 and 2, and a total of 318 samples were tested (152 blood donors, 108 asymptomatic carriers, 26 HAM/TSP patients and 30 seronegative individuals). RESULTS: The sensitivity and specificity of PCR in comparison with WB results were 99.4% and 98.5%, respectively. PCR tests were more efficient for identifying the virus type, detecting HTLV-2 infection and defining inconclusive cases. CONCLUSIONS: Because real-time PCR is sensitive and practical and costs much less than WB, this technique can be used as a confirmatory test for HTLV in blood banks, as a replacement for WB.

Malacological survey of Biomphalaria snails in municipalities along the Estrada Real in the southeast of the State of Minas Gerais, Brazil

INTRODUCTION: The increasing practice of ecotourism and rural tourism in the State of Minas Gerais, Brazil, highlights the importance of studies concerning the occurrence of potential intermediate hosts of Schistosoma mansoni. This study aimed to identify species of Biomphalaria snails in municipalities along the Estrada Real, an important Brazilian tourism project. METHODS: The specimens were collected in different water collections of 36 municipalities along the Estrada Real in the southeast of the State of Minas Gerais. Biomphalaria species were characterized using both morphological and molecular approaches. The research was conducted between August 2005 and September 2009 and all the sites visited were georeferenced using GPS. RESULTS: Six Biomphalaria species were found in 30 of the 36 municipalities studied: glabrata, tenagophila, straminea, peregrina, occidentalis and schrammi. The first three species of Biomphalaria, recognized as intermediate hosts of S. mansoni, were present in 33.3%, 47.2% and 8.3% of the municipalities studied, respectively. The mollusks were found in different types of water collections and no infection by S. mansoni was detected. The highest occurrence of Biomphalaria concentration was verified in the area covered by the Caminho Novo route (Diamantina/MG to Rio de Janeiro/RJ). CONCLUSIONS: Considering the occurrence of schistosomiasis in the State of Minas Gerais and the socioeconomic repercussions involved in the Estrada Real Project, this work focuses on the vulnerability of water collections due to the presence of Biomphalaria mollusks and emphasizes the need for epidemiological surveillance and sanitary and educational measures integrated with the local community and tourism sectors.

Rapid detection of Van genes in rectal swabs by real time PCR in Southern Brazil

INTRODUCTION: Laboratory-based surveillance is an important component in the control of vancomycin resistant enterococci (VRE). METHODS: The study aimed to evaluate real-time polymerase chain reaction (RT-PCR) (genes vanA-vanB) for VRE detection on 115 swabs from patients included in a surveillance program. RESULTS: Sensitivity of RT-PCR was similar to primary culture (75% and 79.5%, respectively) when compared to broth enriched culture, whereas specificity was 83.1%. CONCLUSIONS: RT-PCR provides same day results, however it showed low sensitivity for VRE detection.

Atitudes em relação ao corpo e à alimentação de pacientes com anorexia e bulimia nervosa

Objetivo: Avaliar como se relacionam as atitudes alimentares e corporais de pacientes com anorexia ou bulimia nervosa. Métodos: Pacientes adultas de um ambulatório especializado, com diagnóstico de anorexia (n = 48) ou bulimia nervosa (n = 58), responderam à Disordered Eating Attitude Scale (DEAS) para avaliação das atitudes alimentares e ao Body Attitude Questionnaire (BAQ) para atitudes corporais – ambos traduzidos e validados para mulheres jovens do Brasil. A correlação entre os escores da DEAS e do BAQ foi avaliada utilizando o coeficiente de Pearson. Modelos de regressão linear testaram preditores para atitudes alimentares e corporais. Resultados: Pacientes com bulimia apresentam relação com o alimento mais disfuncional – subescala 1 da DEAS (p < 0,001) e piores atitudes corporais (BAQ total e cinco fatores). Correlações entre a DEAS e BAQ foram mais fortes para a anorexia do que para bulimia e houve forte correlação (r > 0,6) para ambas apenas quando se analisou a relação com o alimento e o sentir-se gorda e entre atitudes corporais como um todo e a relação com o alimento. O escore total da DEAS foi preditor da BAQ total: cada um ponto na DEAS aumenta 0,788 na BAQ (R2 = 0,628). Conclusão: Pacientes com bulimia apresentam pior relação com o alimento e piores atitudes corporais. As atitudes corporais se correlacionaram com as atitudes alimentares, de maneira mais forte para pacientes com anorexia; atitudes alimentares mais disfuncionais predizem pior relação com o corpo para ambos os diagnósticos.

Preocupação com a forma do corpo de graduandos de Farmácia-Bioquímica

Objetivo Identificar a preocupação com a forma do corpo de estudantes de Farmácia-Bioquímica e sua relação com variáveis sociais e laborais e com o estado nutricional. Métodos Participaram 346 discentes com média de idade de 20,2 (DP = 2,4) anos, sendo 278 (80,3%) do sexo feminino. Utilizou-se o Body Shape Questionnaire (BSQ). As validades fatorial e convergente e a consistência interna (α) do BSQ foram estimadas. Utilizaram-se como índices de ajustamento o qui-quadrado pelos graus de liberdade (χ2/gl), o Comparative Fit Index (CFI), o Normed Fit Index (NFI) e o Root Mean Square Error of Approximation (RMSEA). O escore médio de preocupação com a forma do corpo foi obtido por meio de algoritmo gerado na análise fatorial confirmatória. Para comparar os escores médios segundo as variáveis de interesse, utilizou-se Análise de Variância (ANOVA). Resultados O BSQ apresentou, para a amostra de estudo, adequada validade (χ2/gl = 3,29; CFI = 0,87, NFI = 0,82, RMSEA = 0,08) e confiabilidade (α = 0,97) após ajustamento. Verificou-se que as mulheres (p < 0,001) apresentaram maior preocupação com a forma do corpo que os homens. Além disso, os estudantes que avaliaram o curso como pior que as expectativas iniciais (p = 0,048), que consomem medicamentos por causa dos estudos (p < 0,001), que já pensaram em desistir do curso (p = 0,002) e foram classificados com sobrepeso/obesidade (p < 0,001) também apresentaram alta preocupação com a forma do corpo. Conclusão As varáveis sexo, avaliação em relação ao curso, ingestão de medicamentos por causa dos estudos, pensamento em desistir do curso e o estado nutricional apresentaram relação significativa com a preocupação com a forma do corpo entre os estudantes.