Arbovirus infections are widespread, and their disease burden has increased in the past decade. In Africa, arbovirus infections and fever with unknown etiology are common. Due to the lack of well-established epidemiologic surveillance systems and accurate differential diagnosis in most African countries, little is known about the prevalence of human arbovirus infections in Africa. The aim of this review is to summarize the available epidemiological data and diagnostic laboratory tools of infections with dengue, yellow fever, Zika, and chikungunya viruses, all transmitted by Aedes mosquitoes. Studies indicate that these arboviral infections are endemic in most of Africa. Surveillance of the incidence and prevalence of the infections would enable medical doctors to improve the diagnostic accuracy in patients with typical symptoms. If possible, arboviral diagnostic tests should be added to the routine healthcare systems. Healthcare providers should be informed about the prevalent arboviral diseases to identify possible cases.

Les entérovirus (EV) sont des agents étiologiques de nombreuses pathologies chez les adultes et les enfants, y compris la poliomyélite qui a été éradiquée en France. Aujourd’hui, la surveillance d’EV se déroule dans le cadre de la vigilance post-éradication, et fournit des données épidémiologiques importants sur des entérovirus non polio.A Marseille, la surveillance a amené à l’analyse de 654 souches isolées entre 1985 et 2005. Les EV de l'espèce B étaient prédominants, parmi lesquels l’echovirus 30 (E30) a été le plus fréquemment isolé et l’E13 a émergé lors de l’épidémie en 2000. Notre analyse des souches cliniques sur 20 ans renforce la stratégie de sérotypage par la VP1. L'analyse phylogénétique a identifié des souches de différents sérotypes, avec des régions nonstructurales génétiquement proches associées aux VP1 divergents. Ceci contredit le modèle actuel de la recombinaison et pourrait être à l’origine de l'émergence d’E13 épidémique.Deux épidémies majeures d’E30 ont été décrites en 2000 et 2005. Entre elles, le protocole de diagnostic d’EV a été modifié: en 2000, la détection s’est fait par la culture cellulaire et la RT-PCR classique; en 2005, la technique de la RT-PCR en temps réel a été utilisée. Ainsi, l’épidémie de 2005 a été caractérisée par une réduction significative du délai nécessaire de livrer les résultats du diagnostic et de la durée du séjour hospitalier.Nous avons également adapté un système moléculaire pour la détection d'EV71. EV71 est associé à des épidémies du syndrome pied, main, bouche en Asie, mais peut aussi engendrer des complications neurologiques fatales. Nous avons testé 365 échantillons positifs pour l’EV. 3 cas d'EV71 du génogroupe C2 ont été détectés entre 2009 et 2011 chez les enfants sans histoires de voyage récent, ce qui confirme la circulation actuelle de ce génogroupe en France.Les entérovirus ont le potentiel de provoquer des épidémies fréquentes, à cause de leur diversité génétique importante et de leur capacité de réémergence. Il est nécessaire de maintenir la surveillance d'EV par la détection et l'analyse des virus en circulation, et de développer d’autres techniques rapides de détection et d’identification. / Enteroviruses are single-stranded RNA viruses associated with a myriad of pathologies in adult and paediatric populations, the most notable of which, poliomyelitis, has been eradicated in France. Today, enterovirus surveillance is carried out in the context of post-eradication monitoring, and provides important epidemiological data for nonpolio enteroviruses.In Marseille, surveillance efforts culminated in the compilation and analysis of 654 strains isolated between 1985 and 2005. Predominant serotypes belonged to the B species: Echovirus 30 (E30) was the most frequently isolated serotype and E13 emerged during the 2000 epidemic. Our analysis of clinical strains over 20 years lends credence to the VP1 serotyping strategy. Phylogenetic analysis identified strains of different serotypes which were genetically similar in the nonstructural regions despite distinct VP1 regions. This observation contradicts the current model of recombination and could explain the emergence of epidemic E13.Two large outbreaks of E30 were described in 2000 and 2005, in between which EV diagnostic protocol was changed: in 2000, detection was performed using cell culture and classic RT-PCR techniques; in 2005, this was done via real-time RT-PCR. As a result, the 2005 outbreak was characterised by a significant decrease in the time needed to deliver diagnostic results, as well as in the length of hospital stay.We also adapted a real-time molecular assay for the detection of EV71. EV71 is associated with major outbreaks of hand, foot and mouth disease in the Asia-Pacific region, but can also cause fatal neurological complications. We screened 356 EV-positive samples and detected three cases of genogroup C2 EV71 infection between 2009 and 2011 in young children with no history of travel, confirming the current circulation of EV71 in France.Enteroviruses have the potential to cause frequent epidemics, due to their great genetic diversity and their propensity for re-emergence. This underscores the need to maintain EV surveillance by analysing past and present circulating viruses, as well as to develop more rapid detection and identification techniques.

Procedeu-se ao estudo comparativo das técnicas - de Coloração de Sellers, Imunofluorescência Direta e Inoculação em Camundongos, utilizadas na rotina de diagnóstico laboratorial da raiva e praticadas sobre 2.242 amostras de cérebro de cães, enviadas ao Instituto Pasteur de São Paulo, durante o período de janeiro de 1974 a abril de 1976. As técnicas de Inoculação em Camundongos e de Imunofluorescência Direta foram as que apresentaram maior grau de Sensibilidade, sendo essa associação a que forneceu maior Concordância e valor significante mais alto para o x2 (Qui quadrado). A técnica de Coloração de Sellers foi a que apresentou menor grau de Sensibilidade e sua associação com as técnicas de Imunofluorescência Direta e de Inoculação em Camundongos forneceu resultados de Concordância e x2 (Qui quadrado) inferiores aos obtidos pela associação dessas duas técnicas. / A comparative study was made of three techniques: Coloration of Sellers, Direct Immunofluorescence, and Inoculation in Mice - used in the routine of laboratory diagnosis of rabies and applied on 2,242 samples of dog brains which were sent to the Pasteur Institute of Sã0 Paulo, between January 1974 and April 1976. The techniques of mice inoculation and of direct immunofluorescence were the ones which presented higher degree of sensitivity; this association resulted in a greater rapport and a higher significative value for the x2 (square Qui). The techniques of Sellers Coloration resulted in a lower degree of sensitivity, and its association with the techniques of direct immunofluorescence and inoculation in mice gave over results of rapport and x2 than those obtained by the association of the two latter techniques.

Procedeu-se ao estudo comparativo das técnicas - de Coloração de Sellers, Imunofluorescência Direta e Inoculação em Camundongos, utilizadas na rotina de diagnóstico laboratorial da raiva e praticadas sobre 2.242 amostras de cérebro de cães, enviadas ao Instituto Pasteur de São Paulo, durante o período de janeiro de 1974 a abril de 1976. As técnicas de Inoculação em Camundongos e de Imunofluorescência Direta foram as que apresentaram maior grau de Sensibilidade, sendo essa associação a que forneceu maior Concordância e valor significante mais alto para o x2 (Qui quadrado). A técnica de Coloração de Sellers foi a que apresentou menor grau de Sensibilidade e sua associação com as técnicas de Imunofluorescência Direta e de Inoculação em Camundongos forneceu resultados de Concordância e x2 (Qui quadrado) inferiores aos obtidos pela associação dessas duas técnicas. / A comparative study was made of three techniques: Coloration of Sellers, Direct Immunofluorescence, and Inoculation in Mice - used in the routine of laboratory diagnosis of rabies and applied on 2,242 samples of dog brains which were sent to the Pasteur Institute of Sã0 Paulo, between January 1974 and April 1976. The techniques of mice inoculation and of direct immunofluorescence were the ones which presented higher degree of sensitivity; this association resulted in a greater rapport and a higher significative value for the x2 (square Qui). The techniques of Sellers Coloration resulted in a lower degree of sensitivity, and its association with the techniques of direct immunofluorescence and inoculation in mice gave over results of rapport and x2 than those obtained by the association of the two latter techniques.

This thesis deals with the problems of pertussis epidemiological problems in the Czech Republic, especially the possibilities of improvement of the epidemiological situation of pertussis. Pertussis is a serious infectious disease of the respiratory system and is monitored long-term in Czech Republic. The incidence of pertussis in its epidemic cycles continues to increase since the 90s. Last year's reported sickness rate was 11.7 of 100 000 people (1233 cases total).The thesis is divided into theoretical and practical part. The theoretical part shows the characteristics of pertussis and describes the preventive and repressive measures. Last part of the theoretical section describes epidemiological situation of pertussis from history to the present day while last four years are described in more detail. The practical part of the thesis consists of research.The research was conducted using qualitative method. As data collection method was chosen interviews with first contact physicians, which are pediatricians and physicians for adults. Two objectives were set in research area. The first objective was to map the compliance with the conditions laid down under the "case" definition of pertussis and its methods of diagnosis for pediatricians and physicians for adults (first contact physicians). The second objective was to create intervention for affecting the adverse epidemiological situation of pertussis. Intervention is based on the results of research aimed at compliance with procedures of "case" definition of pertussis and on knowledge of pediatricians and physicians for adults about the epidemiology of pertussis issue and their views on vaccination.

Mitochondria are the key source of vital ATP molecules, which are largely produced within cells by a system of oxidative phosphorylation (OXPHOS). Genetic defects affecting any of the components of the oxidative phosphorylation system or the structure and function of mitochondria lead to mitochondrial disorders, which occur at an incidence rate of 1 in 5000 live births. Cytochrome c oxidase (COX) is the terminal enzyme and electron acceptor of a respiratory chain that catalyses oxygen to produce a water molecule. In addition to complex I deficiency, isolated or combined COX deficiency is the most common respiratory chain defect in paediatric patients, and it can arise from mutations located either in mitochondrial DNA or in nuclear genes encoding the structural subunits or corresponding assembly factors of the enzyme complex. However, the molecular basis of COX deficiency remains elusive in many patients despite advances in the identification of an increasing number of mutations and genes involved in the disease. This thesis focuses on the identification of the genetic causes of mitochondrial diseases in a cohort of 60 unrelated Czech children with clinically and laboratory confirmed COX-deficiency. With the use of a high-resolution melting analysis mutation screen, four heterozygous sequence...

Mitochondria are the key source of vital ATP molecules, which are largely produced within cells by a system of oxidative phosphorylation (OXPHOS). Genetic defects affecting any of the components of the oxidative phosphorylation system or the structure and function of mitochondria lead to mitochondrial disorders, which occur at an incidence rate of 1 in 5000 live births. Cytochrome c oxidase (COX) is the terminal enzyme and electron acceptor of a respiratory chain that catalyses oxygen to produce a water molecule. In addition to complex I deficiency, isolated or combined COX deficiency is the most common respiratory chain defect in paediatric patients, and it can arise from mutations located either in mitochondrial DNA or in nuclear genes encoding the structural subunits or corresponding assembly factors of the enzyme complex. However, the molecular basis of COX deficiency remains elusive in many patients despite advances in the identification of an increasing number of mutations and genes involved in the disease. This thesis focuses on the identification of the genetic causes of mitochondrial diseases in a cohort of 60 unrelated Czech children with clinically and laboratory confirmed COX-deficiency. With the use of a high-resolution melting analysis mutation screen, four heterozygous sequence...

This diploma thesis deals with laboratory functional diagnostics of malabsorption syndromes. Especially the functional diagnosis of lactose intolerance. The golden standard for the functional diagnosis of lactose intolerance is the breath test. The aim of the study was to determine the prevalence of lactose intolerance (LI) using a lactose breath test when consuming a dose of 20 g of lactose in patients with gastrointestinal symptoms and in volunteers who do not show gastrointestinal problems. Part of the research was also to examine the symptoms that led the individual to see a doctor and undergo a lactose breath test. The method consisted in the evaluation of the results of breath tests, which were performed on patients and volunteers in the gastroenterological laboratory of the 1st Medical Faculty of Charles University and the General Hospital. A questionnaire was given to everyone to evaluate the symptoms of LI and the phenomena leading to malabsorption. Lactose intolerance was confirmed in 92 % of patients and 79 % of volunteers. The most reported symptoms in the patient sample were typical gastrointestinal LI-related problems: gastric and intestinal problems after milk consumption (85 % of patients), gastric and intestinal problems after consumption of dairy products (85 % of patients),...