Rotavírus têm sido identificados mundialmente como o mais importante agente etiológico de diarréias agudas não-bacterianas em animais jovens de várias espécies, incluindo a humana. Foi desenvolvido e avaliado um método de ELISA tipo “duplo-sanduíche” para a detecção de rotavírus a partir de material fecal. Para tanto, a amostra NCDV de rotavírus do grupo A foi propagada em cultivo celular com células MA-104. O vírus foi concentrado por ultracentrifugação e inoculado em coelhos e carneiros. Em seguida, as frações IgG, oriundas de amostras de soro dos animais, foram purificadas por cromatografia de troca iônica e absorvidas com soro total de ambas espécies animais, utilizando-se polímero de glutaraldeído, de modo a eliminar reações inespecíficas. A presença do rotavírus foi detectada pela IgG de carneiros e revelada pela IgG de coelho, usando como conjugado IgG de cabra anti-IgG de coelho conjugada à peroxidase. Os valores de diluição dos componentes do ELISA e o valor do ponto-de-corte foram definidos usando-se 26 amostras fecais (13 positivas e 13 negativas) de leitões, tendo como prova padrão a eletroforese em gel de poliacrilamida (PAGE). Aplicado a um painel constituído de 86 amostras fecais diarréicas de leitões, os resultados do ELISA foram: 100% de sensibilidade; 98,79% de especificidade, com uma concordância de 98,83%. A variância entre 86 repetições da mesma amostra foram 0,001 (para a amostra positiva) e 0,0002 (para a amostra negativa). Estes resultados demonstram que este ELISA é um teste sensível e específico para o diagnóstico de rotavírus a partir de material fecal. / Rotaviruses have been identified worldwide as a major etiologic agent of acute nonbacterial diarrhea in the young of many species, including humans. In this investigation was developed and evaluated a “double-sandwich” antibody ELISA method for detection of rotavirus from stool specimens. For that, the NCDV strain of rotavirus group A was serially cultivated in MA-104 cell culture. The virus was concentrated by ultra-centrifugation and inoculated in rabbits and sheeps. After that, the IgG of serum samples of the animals was purified by ion-exchange chromatography and absorbed with whole serum of both animal species using a glutaraldehyde polymer, in order to eliminate inespecific reactions. The presence of rotavirus was detected by the sheep’s IgG and revelated by the rabbit’s IgG, using a anti-rabbit IgG peroxidase conjugate developed in goat. The values of diluition of the components of the ELISA and the cut-off value were defined using 26 fecal samples (13 positive and 13 negative) of piglets. Following this procedure, the test was employed in a panel of 86 fecal samples from piglets with diarrhea, using as standard the polyacrilamide gel electrophoresis (PAGE) test. The results of the ELISA were: 100% of sensivity; 98.79% of specificity, with an agreement of 98.83%. The variance between 86 repetitions of the same sample were 0.001 (for one positive sample) and 0.0002 (for one negative sample). These results showed that this ELISA is an sensitive and specific screening test for rotavirus diagnosis from fecal material.

Thesis (Licenciado en Nutriciʹon y Dietʹtica)--Universidad Tʹecnica del Norte. Facultad Ciencias de la Salud Escuela de Nutriciʹon y Dietʹeica. / Abstract in Spanish and English.

Worldwide, diarrheal disease is the second leading cause of death in children under 5 years old. Data on diarrhea of extended duration is limited. We described the characteristics associated with acute, prolonged acute and persistent diarrhea in Kenyan children less than 5 years of age participating in the Global Enterics Multicenter Study. Children presenting at a clinic were enrolled if they met the case definition for acute moderate-to-severe diarrhea defined as >3 loose stools in the last 24 hrs, within 7 days of illness onset, with >1 of the following: sunken eyes, skin tenting, dysentery, IV rehydration, or hospitalization. To determine diarrhea duration, the child’s caretaker was asked to recall the number of days the child had diarrhea in the 7 days pre-enrollment, and to record each day of diarrhea post-enrollment on a form for 14 days. Stool specimens were collected at enrollment, and the post-enrollment form was collected during a home visit. We defined acute diarrhea (AD) as <6 days>duration, prolonged acute diarrhea (ProD) as 7-13 days, and persistent diarrhea (PD) as >14 days. From January 31, 2008 to January 24, 2010, 557 children with acute moderate-to-severe diarrhea were enrolled. Using the Wilcoxon rank-sum test, Kruskal-Wallis test, and Cox Proportional Hazards Model we examined the relationship between the duration of diarrhea by gender, age, and various etiologic agents. We found no association between gender and the duration of diarrhea. Age was associated with diarrhea of extended duration; children less than or equal to 11 months of age were 1.3 times more likely to experience diarrhea of longer duration than their counterparts. We found Cryptosporidium to be more associated with ProAD and PD. Children infected with Cryptosporidium were 1.5 to 1.7 times more likely to have diarrhea with a longer duration than their counterparts. Based on these results, interventions related to diarrhea and diarrhea of extended duration should focus more closely on young children, especially children less than 24 months of age.

Background: Rotavirus is one of the most common diarrheal diseases in children less than 5 years of age. Rotavirus vaccines have greatly reduced this burden in the United States. An examination was conducted to determine possible disparities in RV vaccination rates compared to DTaP. Methods: Children were actively enrolled during two rotavirus seasons from January-June of 2010 and 2011 in the Emergency Departments (ED) and inpatient floors from all Children's Healthcare of Atlanta (CHOA) sites (Scottish Rite, Egleston, and Hughes Spalding) with acute gastroenteritis (AGE). Data and a stool sample were collected from enrolled children and samples were tested for presence of rotavirus using an enzyme immunoassay (EIA) kit (Rotaclone). Vaccination records were abstracted from the state immunization registry and primary healthcare providers to examine complete and incomplete vaccination status. This cohort of children with vaccination records were used for this analysis. Cases were identified as children receiving a complete RV dose series and controls were identified as children with incomplete RV doses. A logistic regression model was used to determine disparities seen amongst children with incomplete vaccination status. Results: Of the 660 patients that were approached for this study, 414 participants were included in this retrospective cohort analysis. 46.9% had incomplete rotavirus vaccination status and were more likely to be positive for rotavirus AGE (OR 1.76, 95% CI 1.46-2.13). Black infants had a higher rate of incomplete RV compared to whites (p-value 0.0006). When controlling for covariates, racial differences were no longer significant (OR 1.37 95% CI 0.77-2.57); however household size (p-value 0.0343), age at onset of illness (p-value 0.0061), and DTaP vaccination status (p-value < 0.0001) were all significant in determining vaccination status for children. Conclusions: Racial disparities and socioeconomic differences are not evident in determining rotavirus vaccination rates; however, household size, a possible social determinant, has an effect on RV status. In addition, timely vaccinations are important in preventing incomplete RV vaccination status, due to RV vaccine age restrictions.

Diss. (sammanfattning)--Linköping : Linköpings universitet, 2006. / Härtill 5 uppsatser.

Thesis--Auburn University, 2009. / Abstract. Vita. Includes bibliographical references (p. 39-44).

Thesis (Ph.D.)--Murdoch University, 2006. / Thesis submitted to the Division of Health Sciences.

Thesis (DR. P.H.)--University of Michigan.

Thesis (M.S.)--Oregon State University, 2009. / Printout. Includes bibliographical references (leaves 54-58). Also available on the World Wide Web.

Thesis (MSc (Veterinary Science))--University of Pretoria, 2001. / Includes bibliographical references.