Molecular diagnosis of strongyloidiasis in tropical areas: a comparison of conventional and real-time polymerase chain reaction with parasitological methods

This study aimed to evaluate the use of conventional polymerase chain reaction (cPCR) and real-time quantitative PCR (qPCR) in the diagnosis of human strongyloidiasis from stool samples in tropical areas. Stool samples were collected from individuals and were determined to be positive for Strongyloides stercoralis (group I), negative for S. stercoralis (group II) and positive for other enteroparasite species (group III). DNA specific to S. stercoralis was found in 76.7% of group I samples by cPCR and in 90% of group I samples by qPCR. The results show that molecular methods can be used as alternative tools for detecting S. stercoralis in human stool samples in tropical areas.

Optimisation of a quantitative polymerase chain reaction-based strategy for the detection and quantification of human herpesvirus 6 DNA in patients undergoing allogeneic haematopoietic stem cell transplantation

Human herpesvirus 6 (HHV-6) may cause severe complications after haematopoietic stem cell transplantation (HSCT). Monitoring this virus and providing precise, rapid and early diagnosis of related clinical diseases, constitute essential measures to improve outcomes. A prospective survey on the incidence and clinical features of HHV-6 infections after HSCT has not yet been conducted in Brazilian patients and the impact of this infection on HSCT outcome remains unclear. A rapid test based on real-time quantitative polymerase chain reaction (qPCR) has been optimised to screen and quantify clinical samples for HHV-6. The detection step was based on reaction with TaqMan® hydrolysis probes. A set of previously described primers and probes have been tested to evaluate efficiency, sensitivity and reproducibility. The target efficiency range was 91.4% with linearity ranging from 10-106 copies/reaction and a limit of detection of five copies/reaction or 250 copies/mL of plasma. The qPCR assay developed in the present study was simple, rapid and sensitive, allowing the detection of a wide range of HHV-6 loads. In conclusion, this test may be useful as a practical tool to help elucidate the clinical relevance of HHV-6 infection and reactivation in different scenarios and to determine the need for surveillance.

A sensitive, specific and reproducible real-time polymerase chain reaction method for detection of Plasmodium vivaxandPlasmodium falciparum infection in field-collected anophelines

We describe a simple method for detection of Plasmodium vivaxand Plasmodium falciparum infection in anophelines using a triplex TaqMan real-time polymerase chain reaction (PCR) assay (18S rRNA). We tested the assay on Anopheles darlingi and Anopheles stephensi colony mosquitoes fed withPlasmodium-infected blood meals and in duplicate on field collected An. darlingi. We compared the real-time PCR results of colony-infected and field collected An. darlingi, separately, to a conventional PCR method. We determined that a cytochromeb-PCR method was only 3.33% as sensitive and 93.38% as specific as our real-time PCR assay with field-collected samples. We demonstrate that this assay is sensitive, specific and reproducible.

Polymerase chain reaction detection of LeishmaniaDNA in skin biopsy samples in Sri Lanka where the causative agent of cutaneous leishmaniasis is Leishmania donovani

Leishmania donovani is the known causative agent of both cutaneous (CL) and visceral leishmaniasis in Sri Lanka. CL is considered to be under-reported partly due to relatively poor sensitivity and specificity of microscopic diagnosis. We compared robustness of three previously described polymerase chain reaction (PCR) based methods to detectLeishmania DNA in 38 punch biopsy samples from patients presented with suspected lesions in 2010. Both, Leishmaniagenus-specific JW11/JW12 KDNA and LITSR/L5.8S internal transcribed spacer (ITS)1 PCR assays detected 92% (35/38) of the samples whereas a KDNA assay specific forL. donovani (LdF/LdR) detected only 71% (27/38) of samples. All positive samples showed a L. donovani banding pattern upon HaeIII ITS1 PCR-restriction fragment length polymorphism analysis. PCR assay specificity was evaluated in samples containing Mycobacterium tuberculosis, Mycobacterium leprae, and human DNA, and there was no cross-amplification in JW11/JW12 and LITSR/L5.8S PCR assays. The LdF/LdR PCR assay did not amplify M. leprae or human DNA although 500 bp and 700 bp bands were observed in M. tuberculosis samples. In conclusion, it was successfully shown in this study that it is possible to diagnose Sri Lankan CL with high accuracy, to genus and species identification, using Leishmania DNA PCR assays.

Commercial enzyme-linked immunosorbent assay versuspolymerase chain reaction for the diagnosis of chronic Chagas disease: a systematic review and meta-analysis

Chronic Chagas disease diagnosis relies on laboratory tests due to its clinical characteristics. The aim of this research was to review commercial enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) diagnostic test performance. Performance of commercial ELISA or PCR for the diagnosis of chronic Chagas disease were systematically searched in PubMed, Scopus, Embase, ISI Web, and LILACS through the bibliography from 1980-2014 and by contact with the manufacturers. The risk of bias was assessed with QUADAS-2. Heterogeneity was estimated with the I2 statistic. Accuracies provided by the manufacturers usually overestimate the accuracy provided by academia. The risk of bias is high in most tests and in most QUADAS dimensions. Heterogeneity is high in either sensitivity, specificity, or both. The evidence regarding commercial ELISA and ELISA-rec sensitivity and specificity indicates that there is overestimation. The current recommendation to use two simultaneous serological tests can be supported by the risk of bias analysis and the amount of heterogeneity but not by the observed accuracies. The usefulness of PCR tests are debatable and health care providers should not order them on a routine basis. PCR may be used in selected cases due to its potential to detect seronegative subjects.

An in-house real-time polymerase chain reaction: standardisation and comparison with the Cobas Amplicor HBV monitor and Cobas AmpliPrep/Cobas TaqMan HBV tests for the quantification of hepatitis B virus DNA

This study aimed to standardise an in-house real-time polymerase chain reaction (rtPCR) to allow quantification of hepatitis B virus (HBV) DNA in serum or plasma samples, and to compare this method with two commercial assays, the Cobas Amplicor HBV monitor and the Cobas AmpliPrep/Cobas TaqMan HBV test. Samples from 397 patients from the state of São Paulo were analysed by all three methods. Fifty-two samples were from patients who were human immunodeficiency virus and hepatitis C virus positive, but HBV negative. Genotypes were characterised, and the viral load was measure in each sample. The in-house rtPCR showed an excellent success rate compared with commercial tests; inter-assay and intra-assay coefficients correlated with commercial tests (r = 0.96 and r = 0.913, p < 0.001) and the in-house test showed no genotype-dependent differences in detection and quantification rates. The in-house assay tested in this study could be used for screening and quantifying HBV DNA in order to monitor patients during therapy.

The identification and differentiation of the Candida parapsilosis complex species by polymerase chain reaction-restriction fragment length polymorphism of the internal transcribed spacer region of the rDNA

Currently, it is accepted that there are three species that were formerly grouped under Candida parapsilosis: C. para- psilosis sensu stricto, Candida orthopsilosis, andCandida metapsilosis. In fact, the antifungal susceptibility profiles and distinct virulence attributes demonstrate the differences in these nosocomial pathogens. An accurate, fast, and economical identification of fungal species has been the main goal in mycology. In the present study, we searched sequences that were available in the GenBank database in order to identify the complete sequence for the internal transcribed spacer (ITS)1-5.8S-ITS2 region, which is comprised of the forward and reverse primers ITS1 and ITS4. Subsequently, an in silico polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed to differentiate the C. parapsilosis complex species. Ninety-eight clinical isolates from patients with fungaemia were submitted for analysis, where 59 isolates were identified as C. parapsilosis sensu stricto, 37 were identified as C. orthopsilosis, and two were identified as C. metapsilosis. PCR-RFLP quickly and accurately identified C. parapsilosis complex species, making this method an alternative and routine identification system for use in clinical mycology laboratories.

Scenarios for the milk production chain in Brazil in 2020

Brazilian milk production has grown steadily and in 2004 the country became self-sufficient in dairy production. This article develops possible scenarios for the milk production chain in Brazil for the year 2020 in order to contribute to decisions that must be made by stakeholders. A literature review on foresight and the use of scenarios was conducted, and a scenario writing approach based on Wright and Spers (2006) was adopted, which includes the use of the Delphi method, Michael Porter's Five Competitive Forces model, Interpretative Structural Modeling (ISM) (WRIGHT, 1991) and quantitative projections. This methodology provided four scenarios, with quantitative and qualitative elements: two exploratory scenarios ("milk, the new agribusiness star" and "a wasted future"), a most probable scenario ("continuous but uneven growth") and a desired scenario ("competitive family agriculture"). Overall, it is possible to note many market opportunities, as well as niche markets and the strengthening of cooperatives. Future prospects are also favorable to the dairy industry in general, but nearly all scenarios point to a concentration in the industrial sphere.

Empreendedorismo feminino e o conflito trabalho-família: estudo de multicasos no setor de comércio de material de construção da cidade de Curitiba

Os autores que analisam o conflito trabalho-família têm ressaltado a importância da busca do equilíbrio do tempo na dedicação para o trabalho sem que haja detrimento do tempo dedicado à família. Destacam-se três dimensões em que se podem encontrar conflitos entre o trabalho e a família: tempo, tensão e comportamento. Nesta pesquisa, tem-se como objetivo apresentar os conflitos trabalho-família percebidos por duas empresárias do setor de comércio de material de construção da cidade de Curitiba, bem como as ações dessas empresárias para lidarem com os conflitos identificados. Por meio de pesquisa qualitativa, do tipo estudo de multicasos, foram entrevistados tanto as empresárias quanto seus parceiros. Os achados da pesquisa corroboram os estudos citados na revisão teórica, sendo o fator tempo o mais frequentemente citado como gerador de conflitos trabalho-família e o controle emocional como a ação mais citada como atenuante desses mesmos conflitos.

Evidências da relação entre a evolução da gestão ambiental e a adoção de práticas de green supply chain management no setor eletroeletrônico brasileiro

Na pesquisa apresentada, teve-se como objetivo verificar se a evolução da gestão ambiental se relaciona positivamente com a adoção de práticas de green supply chain management (GSCM) por empresas do setor eletroeletrônico do Brasil. Para fazer frente ao objetivo exposto, foi realizada uma pesquisa quantitativa, por meio de um levantamento survey, com cem empresas do setor eletroeletrônico brasileiro. Os dados coletados foram processados por meio de técnicas estatísticas descritivas, análise fatorial exploratória e modelagem de equações estruturais. Os resultados mais importantes da pesquisa foram: na amostra, práticas de GSCM orientadas para a recuperação dos investimentos das empresas, como a revenda de materiais inservíveis e outros resíduos, além da adequação à legislação e à auditoria ambiental, obtiveram médias elevadas; e a hipótese de pesquisa (H1) foi confirmada e considerada estatisticamente válida, indicando que a evolução da gestão ambiental influencia a adoção de práticas de GSCM.

Ocorrência de acidente do trabalho por material perfurocortante entre trabalhadores de enfermagem de um hospital universitário da cidade de Campinas, estado de São Paulo

Com o objetivo de estudar aspectos da ocorrência de acidente do trabalho por material perfurocortante entre trabalhadores de enfermagem de um hospital universitário, foram entrevistados 46 trabalhadores que se acidentaram no período de junho a dezembro de 1994. Os resultados forneceram dados importantes que poderão subsidiar programas de prevenção.

Organização do trabalho na unidade centro de material

Este trabalho aborda a Unidade Centro de Material no contexto hospitalar, e os seus aspectos organizacionais, relacionados ao dimensionamento, zoneamento, recursos materiais e humanos.

A enfermagem frente a acidentes de trabalho com material potencialmente contaminado na era do HIV

Realizou-se esta investigação, com os objetivos de identificar a ocorrência de acidentes de trabalho com material pérfuro-cortante potencialmente contaminado, entre enfermeiros e auxiliares de enfermagem de um Hospital de ensino geral do interior do Estado de São Paulo; relacionar a ocorrência destes acidentes com a categoria profissional e anos de trabalho na instituição; identificar o tipo de material pérfuro-cortante causador do acidente e identificar as condutas tomadas após o acidente, pelo acidentado. Para a coleta de dados, através de entrevista individual, utilizou-se um formulário estruturado onde registrava-se, a causa atribuída ao acidente, o período de atuação do profissional acidentado, o objeto causador, a conduta tomada após o acidente. Evidenciou-se que as Precauções Padrão ou as antigas Precauções Universais não são utilizadas como deveriam, conferindo portanto, risco à equipe de enfermagem. Considera-se que esta equipe necessita de orientações específicas através de educação continuada sobre o assunto.

Ensino de enfermagem em nível médio: utilização de estratégia facilitadora com material reciclável

Este estudo teve como objetivo desenvolver estratégias facilitadoras no processo de ensino e aprendizagem, utilizando-se de materiais recicláveis na construção de estruturas e órgãos anatômicos. Foi desenvolvido na disciplina Enfermagem Médico-Cirúrgica, a 31 alunos no curso de nível médio para formação do Auxiliar de Enfermagem. Após leituras de referências bibliográficas relacionadas à Anatomia, Fisiologia e Patologias mais comuns dos diversos sistemas do corpo humano, os alunos selecionaram os materiais recicláveis disponíveis para a confecção das principais estruturas anatômicas, o que possibilitou a visualização das alterações morfológicas decorrentes das afecções, facilitando assim a compreensão das patologias. As estratégias utilizadas favoreceram a aquisição de conhecimentos ao se projetarem em algo concreto, resultado da troca de sugestões e experiências entre os estudantes, favorecido pelo trabalho em grupo.