Using a combination of MLPA kits to detect chromosomal imbalances in patients with multiple congenital anomalies and mental retardation is a valuable choice for developing countries

Conventional karyotyping detects anomalies in 3-15% of patients with multiple congenital anomalies and mental retardation (MCA/MR). Whole-genome array screening (WGAS) has been consistently suggested as the first choice diagnostic test for this group of patients, but it is very costly for large-scale use in developing countries. We evaluated the use of a combination of Multiplex Ligation-dependent Probe Amplification (MLPA) kits to increase the detection rate of chromosomal abnormalities in MCA/MR patients. We screened 261 MCA/MR patients with two subtelomeric and one microdeletion kits. This would theoretically detect up to 70% of all submicroscopic abnormalities. Additionally we scored the de Vries score for 209 patients in an effort to find a suitable cut-off for MLPA screening. Our results reveal that chromosomal abnormalities were present in 87 (33.3%) patients, but only 57 (21.8%) were considered causative. Karyotyping detected 15 abnormalities (6.9%), while MLPA identified 54 (20.7%). Our combined MLPA screening raised the total detection number of pathogenic imbalances more than three times when compared to conventional karyotyping. We also show that using the de Vries score as a cutoff for this screening would only be suitable under financial restrictions. A decision analytic model was constructed with three possible strategies: karyotype, karyotype + MLPA and karyotype + WGAS. Karyotype + MLPA strategy detected anomalies in 19.8% of cases which account for 76.45% of the expected yield for karyotype + WGAS. Incremental Cost Effectiveness Ratio (ICER) of MLPA is three times lower than that of WGAS, which means that, for the same costs, we have three additional diagnoses with MLPA but only one with WGAS. We list all causative alterations found, including rare findings, such as reciprocal duplications of regions deleted in Sotos and Williams-Beuren syndromes. We also describe imbalances that were considered polymorphisms or rare variants, such as the new SNP that confounded the analysis of the 22q13.3 deletion syndrome. (C) 2011 Elsevier Masson SAS. All rights reserved.

FALSE-NEGATIVE DENGUE CASES IN RORAIMA, BRAZIL: AN APPROACH REGARDING THE HIGH NUMBER OF NEGATIVE RESULTS BY NS1 AG KITS

Serum samples from 150 NS1-negative (Platelia ELISA) patients presumptively diagnosed with dengue were analyzed by the TaqMan probed real-time reverse transcription PCR (TaqMan qRT-PCR) method. The qRT-PCR positive samples were tested for serotype by semi-nested RT-PCR and a qualitative immunochromatographic assay for IgG and IgM. Molecular detection methods showed 33 (22%) positive samples out of 150 NS1-antigen negative samples. Of these, 72% were collected up to day 2 after the onset of symptoms, when diagnostic sensitivity of NS1-antigen test assays is significantly enhanced. Most of the cases were not characterized as secondary infection. Twenty-eight samples were successfully serotyped, 75% of which for DENV-4, 14% for DENV-2, 7% for DENV-3 and 4% for DENV-1. These findings reaffirm the hyperendemic situation of the state of Roraima and suggest a lower sensitivity of the NS1 test, mainly when DENV-4 is the predominant serotype. Health care providers should therefore be aware of samples tested negative by NS1 antigen assays, especially when clinical symptoms and other laboratory data results show evidence of dengue infection.

Evaluation of Lionex TB kits and mycobacterial antigens for IgG and IgA detection in cerebrospinal fluid from tuberculosis meningitis patients

To evaluate commercial Lionex TB together with four antigens of Mycobacterium tuberculosis (MPT-64, MT10.3, 16 kDa and 38 kDa) for IgG and IgA cerebrospinal fluid (CSF) detection in the diagnosis of tuberculosis meningitis (TBM) with CSF negative acid-fast bacilli staining, 19 cases of TBM, 64 cases of other infectious meningoencephalitis and 73 cases of other neurological disorders were tested by enzyme linked immunosorbent assay. IgA-MPT-64 and IgG Lionex showed the highest sensitivities, specificities, positive predictive value and negative predictive value (63.2%, 47.4%; 95%, 93.7%; 40%, 98% and 28.4%, 97.1%, respectively). However, while grey zone was 12.7% and 6%, respectively, lowering sensitivity but maintains high specificity (> 95%). High protein concentration in CSF was associated with antibody positivity CSF/HIV+ which did not influence the sensitivity of both tests. To our knowledge, this is the first description of IgA-MPT-64 and IgG Lionex antibodies in CSF-TBM and, although there is good specificity, adjustments are needed based on antigen composition to enhance sensitivity.

Kits para aerossol em um serviço de saúde: uma análise microbiológica após reprocessamento

Kits para aerossol são artigos utilizados na terapêutica de afecções do trato respiratório e requerem no mínimo desinfecção de nível intermediário para reuso. Os objetivos deste estudo foram verificar uma possível contaminação microbiana em kits para aerossol pós-reprocessamento e identificar os microrganismos isolados. Estudo transversal, experimental realizado na unidade pediátrica de um hospital em Goiânia-GO. Coletaram-se amostras de três segmentos (máscara, copo, interior da extensão) de 15 kits previamente desinfetados, que foram semeadas em diferentes meios de culturas e os microrganismos isolados foram identificados por provas bioquímicas. Dos 15 kits analisados, 13 copos, nove extensões e 13 máscaras estavam contaminados. Isolou-se no total 101 UFC, 39 provindos dos copos, 20 das extensões e 42 das máscaras. Dentre os patógenos isolados destaca-se: Staphylococcus coagulase positivo, Staphylococcus coagulase negativo, Bastonetes Gram negativo fermentadores, Bastonetes Gram negativo não fermentadores, micrococose leveduras. A detecção microbiana indica prováveis falhas no reprocessamento desses artigos.

Comparison of two commercial kits and two extraction methods for fecal glucocorticoid analysis in ocelots (Leopardus pardalis) submitted to ACTH challenge

The ocelot (Leopardus pardalis) is included in list of wild felid species protected by CITES and is part of conservation strategies that necessarily involve the use of assisted reproduction techniques, which requires practical and minimally invasive techniques of high reproducibility that permit the study of animal reproductive physiology. The objective of this study was to compare and validate two commercial assays: ImmuChem Double Antibody Corticosterone 125I RIA from ICN Biomedicals, Costa Mesa, CA, USA; and Coat-a-Count Cortisol 125I RIA from DPC, Los Angeles, CA, USA, for assessment of fecal glucocorticoid metabolites in ocelots submitted to ACTH (adrenocorticotropic hormone) challenge. Fecal samples were collected from five ocelots kept at the Brazilian Center of Neotropical Felines, Associação Mata Ciliar, São Paulo, Brazil, and one of the animals was chosen as a negative control. The experiment was conducted over a period of 9 days. On day 0, a total dose of 100 IU ACTH was administered intramuscularly. Immediately after collection the samples were stored at 20C in labeled plastic bags. The hormone metabolites were subsequently extracted and assayed using the two commercial kits. Previously it was performed a trial with the DPC kit to check the best extraction method for hormones metabolites. Data were analyzed with the SAS program for Windows V8 and reported as means ± SEM. The Schwarzenberger extraction method was slightly better when compared with the Wasser extraction method (103,334.56 ± 19,010.37ng/g of wet feces and 59,223.61 ± 12,725.36ng/g of wet feces respectively; P=0,0657). The ICN kit detected an increase in glucocorticoid metabolite concentrations in a more reliable manner. Metabolite concentrations (ng/g wet feces) on day 0 and day 1 were 66,956.28 ± 36,786.93 and 92,991.19 ± 28,555.63 for the DPC kit, and 205,483.32 ± 83,811.32 and 814,578.75 ± 292,150.47 for the ICN kit, respectively. The limit of detection for the ICN kit was 7.7 ng/mL for 100% B/Bo (25ng/mL for 88%B/Bo) and for the DPC kit it was 0.2ug/dL for 90.95% B/Bo (1ug/dL for 81.27% B/Bo). In conclusion it was confirmed that the Schwarzenberger extraction method and the ICN kit are superior for extracting and measuring fecal glucocorticoid metabolites in ocelot fecal samples.

Comparação de kits ELISA® comerciais para anticorpos no soro e leite com um teste coproparasitológico em bovinos naturalmente infectados por Fasciola hepatica

A fasciolose é uma enfermidade causada por um trematoda que acomete o fígado principalmente de ruminantes domésticos, podendo parasitar o homem e seu diagnóstico é realizado rotineiramente por exames coproparasitológicos. O objetivo do presente estudo foi comparar kits comerciais de ELISA para anticorpos no soro e leite com um teste coproprarasitológico em bovinos naturalmente infectados por Fasciola hepatica. Foram coletadas amostras de fezes (92) sangue (92) e leite (43) de bovinos provenientes de propriedades de gado leiteiro do município de Jerônimo Monteiro, sul do Estado do Espírito Santo. As amostras de fezes coletadas foram processadas pela técnica de sedimentação fecal para ovos de F. hepatica, utilizada como padrão ouro para as análises. Amostras de sangue e de leite foram processadas segundo a orientação do fabricante dos respectivos Kits ELISA comerciais testados. Utilizou-se o c² de McNemar para comparação estatística e calcularam-se a sensibilidade e especificidade, valores preditivos e kappa. Os resultados obtidos mostraram que as frequências de positividade pelo uso dos kits ELISA comerciais de soro e de leite diferiram significativamente (p<0,0001) em relação ao exame coproparasitológico. A sensibilidade dos Kits foi de 100%, porém possuíram baixa especificidade, 42,85 e 30% para o soro e leite respectivamente. O coeficiente de kappa mostrou concordância sofrível para os testes de soro (0,33) e de leite (0,21). Os valores preditivos positivos dos kits para soro e leite foram, respectivamente, 44,61 e 38,23% e, os valores preditivos negativos de 100% para ambos os testes. Apesar da maior sensibilidade dos kits ELISA comerciais e, destes apresentarem diferença em relação ao exame coproparasitológico na detecção dos animais positivos para F. hepatica, a escolha de um teste diagnóstico deve considerar o custo benefício. Quando se trata da presença de parasitismo em rebanhos, o tratamento é aplicado em todos os animais e, assim, o exame coproparasitológico para o diagnóstico da doença tem maior eficiência, já que é menos oneroso e de fácil execução.

Avaliação da eficiência de dois kits comerciais para detecção de aflatoxina B1 em amostras de milho, ração e amendoim e seus produtos

Foi avaliada a eficiência de dois kits ELISA disponíveis no mercado para detecção de aflatoxina B1, que permitem a triagem de milho contaminado com aflatoxina B1 aos níveis de 5 e 20 ppb. Eles foram comparados ao método de Soares & Rodriguez-Amaya que utiliza cromatografia em camada delgada (CCD) e tem como limite de quantificação 4,0μg/kg (ppb). Foram analisadas 53 amostras incluindo milho, ração para peixe e amendoim e produtos. Os resultados mostraram boa concordância entre as duas técnicas quanto à detecção de aflatoxina B1. O método ELISA mostrou-se mais rápido, menos trabalhoso, utilizando menos solventes orgânicos, porém apresentou resultados presuntivos positivos, tornando necessária a sua confirmação.

Immunoassays, methods for carrying out immunoassays, immunoassay kits and method for manufacturing immunoassay kits

The invention relates to immunoassays, methods for carrying out immunoassays, immunoassay kits and methods for manufacturing immunoassay kits. In particular, the invention has relevance to capillary (especially microcapillary) immunoassay technology.

March 2008: Teaching Kits

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Gerenciamento do risco sanitário hospitalar na área de tecnovigilância: análise retrospectiva e prospectiva das notificações de queixas técnicas, incidentes e eventos adversos relacionados ao uso de equipamentos, materiais médico-hospitalares e kits diagnósticos no Hospital das Clínicas da Faculdade de Medicina de Botucatu-UNESP

Pós-graduação em Bases Gerais da Cirurgia - FMB

A Qualitative Analysis of Youth Feedback of Nutrition School Enrichment Kits in Lincoln, NE

A Qualitative Analysis of Youth Feedback of Nutrition School Enrichment Kits in Lincoln, NE Richard A. Losey, M.S. University of Nebraska, 2010 Advisor: Wanda M. Koszewski The qualitative analysis of the responses given by students participating in classrooms that used the Nutrition Education Program‟s (NEP) school enrichment kits was the primary focus of this research. Data was collected from the participants in written form. Three major themes appeared during analysis of the data, healthy eating habits, cleanliness and change. The theme of healthy eating habits is comprised of the following sub-themes: breakfast, nutrients, label reading, Food Guide Pyramid/MyPyramid and healthy snacks. Cleanliness is comprised of hand washing and food safety sub-themes. The change theme is made up of change in healthy eating habits, change in cleanliness and change in both healthy eating habits and cleanliness. The data suggests that participating students have made or intend to make changes regarding healthy eating habits and cleanliness, which is the goal of nutrition education. The students‟ responses indicate that the NEP school enrichment kits are effective in increasing the knowledge of students and promoting healthy lifestyle changes.