Avaliação de testes imunológicos para o diagnóstico da neurocisticercose

Introdução: O diagnóstico da neurocisticercose (NCC) deve ser feito pela associação de técnicas de imagem com métodos imunológicos sensíveis e específicos. Objetivos: Avaliar os métodos Elisa e Western blot (Wb), utilizando-se como antígeno extrato bruto salino da larva da Taenia solium, o Cysticercus cellulosae e Wb, empregando-se como antígeno cisticercos da Taenia crassiceps em amostras de soro, para o diagnóstico da NCC. Materiais e métodos: Foram avaliadas amostras de soro de 43 pacientes com diagnóstico de NCC: 21 por clínica, tomografia computadorizada de crânio (TC) e presença de anticorpos anticisticerco no líquido cefalorraquiano (LCR); 22 por clínica e TC e 229 pacientes com diferentes parasitoses. Para as análises desses materiais biológicos foram empregados os métodos Elisa, usando-se como antígeno C. cellulosae, e Wb, usando-se como antígeno C. cellulosae e Cysticercus longicollis. Resultados: O método Elisa utilizando C. cellulosae como antígeno apresentou especificidade de 95% e sensibilidade de 71%. O método Wb utilizando C. cellulosae ou C. longicollis como antígeno apresentou sensibilidade de 86% e especificidade de 99%. Conclusões: Os métodos imunológicos no LCR são importantes para a definição da NCC. Entretanto o método Elisa no soro ainda não é adequado pela sua baixa sensibilidade, mas o Wb apresentou alta especificidade e boa sensibilidade, podendo auxiliar no diagnóstico da NCC, possibilitando sugerir a existência de forma transicional da doença, não demonstrada pela TC.<br>Background: The diagnosis of neurocysticercosis (NCC) has been made by association of neuroimaging studies and use of sensitive and specific serological assays. Objectives: Evaluating Elisa and Western blot (Wb) tests using a crude extract of Cysticercus cellulosae (Taenia solium) as antigen and a Wb test using a glycoprotein of Cysticercus longicollis (Taenia crassiceps) as antigen for the diagnosis of NCC. Methods: Serum samples from 43 patients with NCC: 21patients presenting clinical manifestations, cerebral computed tomography findings compatible with cerebral lesions and high levels of anti-cysticercus antibodies in cerebrospinal fluid (CSF); 22 patients with clinical manifestations and cerebral computed tomography findings compatible with cerebral lesions and serum samples from 229 patients with other parasitic infections were tested by Elisa standardized with crude extract of C. cellulosae and Wb standardized with glicoprotein of C. cellulosae and C. longicollis. Results: The Elisa test using crude extract of C. cellulosae showed specificity of 95% and sensibility of 71%. Both tests using glycoproteins of either C. cellulosae or C. longicollis showed specificity of 99% and sensibility of 86%. Conclusions: The use of immunological techniques for antibodies detection in CSF was shown to be an important tool for NCC diagnosis. However, detection of antibodies in serum lacked sensibility when Elisa was evaluated. The Wb assay in serum samples was sensitive and specific and it can be helpful for the diagnosis of the transitional form of NCC frequently not detected by cerebral computed tomography

Differential Diagnosis of Entamoeba

Amoebiasis, a disease caused by Entamoeba histolytica, is usually diagnosed by microscopic examination, which does not differentiate the morphologically identical species of the E. histolytica/E. dispar complex. Furthermore, morphologically similar species such as Entamoeba hartmanni contribute to misidentification. Therefore, there is a need for more sensitive and specific methods. This study standardized a multiplex real-time PCR system for E. histolytica and E. dispar and a single real-time PCR for E. hartmanni. The multiplex protocol detected up to 0.0143 pg of E. histolytica DNA and 0.5156 pg of E. dispar DNA, and the average melting temperature (Tm) was 73°C and 70°C, respectively. For E. hartmanni, the Tm was 73°C and the amplification was successful down to 0.03 fg of plasmid DNA. Negative controls and other intestinal parasites presented no amplification. Among the 48 samples tested, E. dispar DNA was detected in 37; none exhibited E. histolytica DNA and 11 were negative in the multiplex protocol. In 4 of these 11 samples, however, E. hartmanni DNA was amplified. SYBR Green is demonstrated to be an interesting option and these combined PCR reactions can improve laboratory diagnosis of amoebiasis in developing countries

Differential Diagnosis of Entamoeba spp. in Clinical Stool Samples Using SYBR Green Real-Time Polymerase Chain Reaction

Amoebiasis, a disease caused by Entamoeba histolytica, is usually diagnosed by microscopic examination, which does not differentiate the morphologically identical species of the E. histolytica/E. dispar complex. Furthermore, morphologically similar species such as Entamoeba hartmanni contribute to misidentification. Therefore, there is a need for more sensitive and specific methods. This study standardized a multiplex real-time PCR system for E. histolytica and E. dispar and a single real-time PCR for E. hartmanni. The multiplex protocol detected up to 0.0143 pg of E. histolytica DNA and 0.5156 pg of E. dispar DNA, and the average melting temperature ( ) was 73 ∘ C and 70 ∘ C, respectively. For E. hartmanni, the was 73 ∘ C and the amplification was successful down to 0.03 fg of plasmid DNA. Negative controls and other intestinal parasites presented no amplification. Among the 48 samples tested, E. dispar DNA was detected in 37; none exhibited E. histolytica DNA and 11 were negative in the multiplex protocol. In 4 of these 11 samples, however, E. hartmanni DNA was amplified. SYBR Green is demonstrated to be an interesting option and these combined PCR reactions can improve laboratory diagnosis of amoebiasis in developing countries

Rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART): Study protocol for a randomized controlled trial

Background: Acute respiratory distress syndrome (ARDS) is associated with high in-hospital mortality. Alveolar recruitment followed by ventilation at optimal titrated PEEP may reduce ventilator-induced lung injury and improve oxygenation in patients with ARDS, but the effects on mortality and other clinical outcomes remain unknown. This article reports the rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART). Methods/Design: ART is a pragmatic, multicenter, randomized (concealed), controlled trial, which aims to determine if maximum stepwise alveolar recruitment associated with PEEP titration is able to increase 28-day survival in patients with ARDS compared to conventional treatment (ARDSNet strategy). We will enroll adult patients with ARDS of less than 72 h duration. The intervention group will receive an alveolar recruitment maneuver, with stepwise increases of PEEP achieving 45 cmH(2)O and peak pressure of 60 cmH2O, followed by ventilation with optimal PEEP titrated according to the static compliance of the respiratory system. In the control group, mechanical ventilation will follow a conventional protocol (ARDSNet). In both groups, we will use controlled volume mode with low tidal volumes (4 to 6 mL/kg of predicted body weight) and targeting plateau pressure &lt;= 30 cmH2O. The primary outcome is 28-day survival, and the secondary outcomes are: length of ICU stay; length of hospital stay; pneumothorax requiring chest tube during first 7 days; barotrauma during first 7 days; mechanical ventilation-free days from days 1 to 28; ICU, in-hospital, and 6-month survival. ART is an event-guided trial planned to last until 520 events (deaths within 28 days) are observed. These events allow detection of a hazard ratio of 0.75, with 90% power and two-tailed type I error of 5%. All analysis will follow the intention-to-treat principle. Discussion: If the ART strategy with maximum recruitment and PEEP titration improves 28-day survival, this will represent a notable advance to the care of ARDS patients. Conversely, if the ART strategy is similar or inferior to the current evidence-based strategy (ARDSNet), this should also change current practice as many institutions routinely employ recruitment maneuvers and set PEEP levels according to some titration method.Hospital do Coracao (HCor) as part of the Program 'Hospitais de Excelencia a Servico do SUS (PROADI-SUS)'Brazilian Ministry of Healt

Blastocystis sp. in splenic cysts: causative agent or accidental association? A unique case report

This work was financed by PAPES-VI/FIOCRUZ. The funders had no participation in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.Made available in DSpace on 2015-04-14T12:38:09Z (GMT). No. of bitstreams: 2 license.txt: 1914 bytes, checksum: 7d48279ffeed55da8dfe2f8e81f3b81f (MD5) sattamini_anaetal_IOC_2014.pdf: 9178458 bytes, checksum: 4ce1e679c31a9ef76f14b3610de46b7b (MD5) Previous issue date: 2014Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Avaliação e Promoção da Saúde Ambiental. Rio de Janeiro, RJ, Brasil.Universidade Federal Fluminense. Departamento de Patologia. Niterói, RJ, Brasil.Universidade Federal Fluminense. Departamento de Patologia. Niterói, RJ, Brasil.Background: Blastocystis sp. is one of the most prevalent parasites found in human stool and has been recently considered an opportunistic emerging pathogen in immunocompromised individuals. However, cases of invasive intestinal infections and skin rashes have been attributed to infection by Blastocystis sp in immunocompetent individuals, suggesting that it is an emerging parasite with pathogenic potential. Findings: We present a case of a 22 year old female patient who complained of pain in the left hypochondrium. Ultrasonography and abdominal computed tomography scans showed two splenic cysts. The cyst fluid analysis demonstrated numerous Blastocystis sp.; PCR and DNA sequencing analyses confirmed the presence of Blastocystis subtype 3. Conclusions: This is, to our knowledge, the first case report of the presence of Blastocystis subtype 3 in extra-intestinal organs and is strong evidence that Blastocystis sp. is potentially pathogenic and invasive. However, further studies are required to determine the pathogenicity of the parasite