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Projektet har haft som syfte att i samarbete med Örsjö Belysning AB ta fram en vägghängd armatur för hotell som ska ha funktion som ljus till att läsa vid sängen. Lampan har haft krav på hållbar utveckling. Lampan har en energieffektiv ljuskälla, vald med hänsyn till sitt syfte, och ingående material har valts med stark miljöhänsyn. Form och funktion har varit viktiga begrepp och har parallellt följts åt i projektet. Produkten har konstruerats för en okomplicerad isärtagning så att delarna enkelt kan sorteras och återvinnas vid armaturens avslutade användning. / The project has served to, in cooperation with Örsjö Belysning AB, develop a wall mounted luminaire for hotel rooms that will function as a bedside reading light. The lamp has had requirements for sustainable development. The lamp has an energy-efficient light source, selected for its purpose, and the materials have been selected by strong environmental concerns. Form following function is an important concept for this project. The product is designed for a straightforward disassembly so that parts can be easily sorted and recycled at the end of use of the fixture.

O Zika vírus (ZIKV) é um vírus RNA de fita única, pertencente à família Flaviviridae. É transmitido entre os humanos geralmente pelos mosquitos da espécie Aedes, mas transmissão via sexual, perinatal e por transfusão sanguínea também foram relatadas. Os sintomas aparecem em 20% dos indivíduos infectados e incluem febre, dor de cabeça, rash cutânea, conjuntivite, mialgia e artralgia. Em 2016, durante a grande epidemia do ZIKV pelas Américas, o interesse pelo seu diagnóstico rápido se intensificou, devido a relação do vírus com o aumento da incidência de casos da síndrome de Guillain-Barré em adultos e da microcefalia em recém nascidos de mulheres grávidas infectadas. De acordo com o CDC (Center for Disease Control and Prevention) o diagnóstico dos pacientes sintomáticos deve ser realizado através da detecção dos ácidos nucleicos do vírus por PCR (Polymerase chain reaction) em amostras pareadas de sangue e urina. Estudos recentes têm postulado que a saliva é uma alternativa importante para detecção do ZIKV. A saliva requer menor complexidade no processamento quando comparada ao sangue, simplificando a reação. A amplificação Isotérmica mediada por Loop (LAMP) é um teste sorológico de alta sensibilidade e especificidade para detectar rapidamente DNA ou RNA de patógenos, incluindo o ZIKV. O fato de não requerer ciclos térmicos como o PCR, faz do LAMP uma reação mais simples, rápida e mais econômica por exigir menos energia. O objetivo deste estudo foi de avaliar e comparar a eficácia da saliva e da urina em diagnosticar a infecção pelo Zika vírus em indivíduos na fase aguda da doença, através da detecção do RNA viral por meio do LAMP. Ao todo, 131 amostras (68 saliva e 63 urina) de 69 indivíduos brasileiros apresentando sinais e sintomas específicos e confirmados positivamente para o ZIKV através da análise do sangue por PCR, foram coletadas e analisadas por LAMP. A média de idade dos indivíduos foi de 34,7 (±13,6), sendo 46 (66,7%) do sexo feminino. Das 68 amostras de saliva analisadas por LAMP, 45 (66,2%) foram positivas para o ZIKV com o Tempo de positividade (Tp) médio de 13,5 minutos. Enquanto que das 63 amostras de urina, 25 (39,7%) foram positivas com o Tp médio de 15,8 minutos. A saliva pôde diagnosticar mais indivíduos (p=0.0042) e em menor Tp (p=0.0176) quando comparada à urina. A saliva demonstrou ser uma alternativa viável no diagnóstico da infecção do ZIKV, em indivíduos na fase aguda da doença, através do LAMP. Nossos achados contribuem para o conhecimento do comportamento do Zika vírus no organismo, uma vez que pouco se conhece em relação à excreção do ZIKV na saliva. / Zika virus (ZIKV) is a single-stranded RNA virus, member of the Flaviviridae family. It is transmitted among humans usually by Aedes mosquito species, but sexual transmission, perinatal and blood transfusion have also been reported. Symptoms appear in 20% of infected individuals and include fever, cutaneous rash, headache, conjunctivitis, myalgia and arthralgia. In 2016, during the Americas ZIKV outbreak, the interest in a rapid diagnosis intensified due to a sudden increase in cases of Guillain-Barré syndrome in adults and microcephaly in newborns of infected pregnant women related with ZIKV. According to CDC (Center for Disease Control and Prevention) the diagnosis of symptomatic patients should be done through nucleic acid detection by PCR (Polimerase Chain Reaction) in paired samples of blood and urine. Recent studies have reported that saliva can be an important alternative to detect ZIKV. Saliva requires less processing than blood, which greatly simplifies the assay process. Loop-mediated Isothermal Amplification (LAMP) is a molecular test with high sensibility and specificity for rapid detection of DNA or RNA of pathogens, including ZIKV. The fact that LAMP does not require thermal cycling makes the assay simple, fast and cost effective compared to PCR assay. The aim of this study was to evaluate the efficacy of saliva and urine to diagnose ZIKV infection in subjects during the acute phase, through ZIKV RNA detection by LAMP. A total of 131 samples (68 saliva and 63 urine) from 69 subjects in the acute phase of ZIKV infection and confirmed positive for ZIKV by blood analysis through PCR were collected and analyzed by LAMP. The mean age of the individuals was 34.7 (±13,6) years old, of whom 46 (66.7%) were females. From the 68 saliva samples, 45 (66.2%) were positive for ZIKV with an average time to positivity (Tp) of 13.5 minutes, and from the 63 urine samples, 25 (39.7%) were positive with an average Tp of 15.8 minutes. Saliva detected more samples (p=0.0042) and had faster Tp (p=0.0176) as compared to urine. Thus, saliva proved to be a feasible alternative for diagnosis of ZIKV infection during the acute phase by LAMP. The findings of this study can contribute to the knowledge of the Zika virus behavior in the human organism, since this issue is not totally understood.

<p>Projektet har haft som syfte att i samarbete med Örsjö Belysning AB ta fram en vägghängd armatur för hotell som ska ha funktion som ljus till att läsa vid sängen. Lampan har haft krav på hållbar utveckling. Lampan har en energieffektiv ljuskälla, vald med hänsyn till sitt syfte, och ingående material har valts med stark miljöhänsyn. Form och funktion har varit viktiga begrepp och har parallellt följts åt i projektet. Produkten har konstruerats för en okomplicerad isärtagning så att delarna enkelt kan sorteras och återvinnas vid armaturens avslutade användning.</p> / <p>The project has served to, in cooperation with Örsjö Belysning AB, develop a wall mounted luminaire for hotel rooms that will function as a bedside reading light. The lamp has had requirements for sustainable development. The lamp has an energy-efficient light source, selected for its purpose, and the materials have been selected by strong environmental concerns. Form following function is an important concept for this project. The product is designed for a straightforward disassembly so that parts can be easily sorted and recycled at the end of use of the fixture.</p>

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2017-02-06T12:12:53Z No. of bitstreams: 2 Tese - Vanessa Duarte Dias - 2016.pdf: 2283005 bytes, checksum: 467160bb33424613a588c7251fabc187 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-02-06T12:13:29Z (GMT) No. of bitstreams: 2 Tese - Vanessa Duarte Dias - 2016.pdf: 2283005 bytes, checksum: 467160bb33424613a588c7251fabc187 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-02-06T12:13:30Z (GMT). No. of bitstreams: 2 Tese - Vanessa Duarte Dias - 2016.pdf: 2283005 bytes, checksum: 467160bb33424613a588c7251fabc187 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-06-30 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The sugarcane production areas are increasing in Brazil due to increased ethanol consumption by flex fuel cars. The planted area is growing, but productivity has been declining in recent years, and factors such as incidence of diseases in the crop may be contributing to this situation. Among the diseases, bacteria such as scald of the leaves and ratoon stunting disease are of great importance to the crop because they can reduce productivity by up to 30%. In addiction and the symptoms are not always displayed in the field, thus requiring advanced techniques to detect such bacterial diseases For Leifsonia xyli subsp. xyli, which causes rickets in the ratoon cane sugar control measure most commonly used besides varietal resistance is thermally treating the billets that will serve seedlings. Thus, the purpose of the first study was to perform the heat water treatment of billets with the addition of kasugamycin antibiotic dosage 300mL/100L H2O in order to try to reduce bacterial escape from the standard treatments, as also in other time and temperature combinations proposed: T1 = 52°C/30 '; T2 = 52°C/1hr; T3 = 50°C/1hr; T4 = 50°C/2hrs; T5 = 52°C/30 '+ antibiotic; T6 = 52°C/1hr + antibiotic; T7 = 50°C/1hr + antibiotic; T8 = 50°C/2hrs + antibiotic; T9 = antibiotic and T10 = control. Moreover, among techniques for diagnosis of such diseases, the most used by laboratories are the serological tests which have the advantage of quantitatively detecting the presence of bacteria on the stems but, however, has the disadvantage of detecting only when the bacterial population is relatively high. PCR technique that is one of the techniques considered most sensitive, has not been used in practice, as this high sensitivity has not been used, the protocols do not detect bacterial diseases in the case of latent infection, where bacterial title is relatively low. So the other work aimed at improving the Xanthomonas albilineans detection developing a LAMP protocol, and compared to other techniques of detection and isolation in semiselective medium, PCR and nested PCR for both symptomatic samples and for asymptomatic and the latter where population in general pathogen is low, we improved Nested one protocol to detect the leaf scald in latent infections by comparing the vascular fluid extraction techniques combined with four different DNA extraction protocols. / O cultivo de cana-de-açúcar está em expansão no Brasil, devido principalmente ao crescente consumo de etanol por carros bicombustíveis. A área plantada está em crescimento, mas a produtividade nos últimos anos vem decrescendo, e fatores como incidência de doenças podem estar contribuindo para tal situação. Dentre as doenças, as bacterianas, como o raquitismo das soqueiras e a escaldadura das folhas, possuem grande importância para a cultura, pois podem reduzir a produtividade em até 30%. Nem sempre os sintomas são visualizados em campo, necessitando assim de técnicas avançadas de detecção ou controle. Para a bacteria Leifsonia xyli subsp. xyli (Lxx), que causa o raquitismo das soqueiras em cana-de-açúcar a medida de controle mais utilizada além da resistência varietal é tratar termicamente os toletes que servirão de mudas. Assim o objetivo do primeiro trabalho foi realizar o tratamento térmico dos toletes com a adição de antibiótico casugamicina à dosagem de 300mL/100L de H2O com o intuito de tentar reduzir o escape bacteriano dentre os tratamentos padrões, como também em outras combinações de tempo e temperatura propostos: T1= 52ºC/30’; T2= 52ºC/1hr; T3= 50ºC/1hr; T4= 50ºC/2hrs; T5= 52ºC/30’ + antibiótico; T6= 52ºC/1hr + antibiótico; T7= 50ºC/1hr + antibiótico; T8= 50ºC/2hrs + antibiótico; T9= antibiótico e T10= testemunha. Além disso, dentre as técnicas para diagnose de tais doenças, as mais utilizadas por laboratórios são os testes sorológicos, que tem como vantagem detectar quantitativamente a presença das bactérias nos colmos mas, no entanto, possui a desvantagem de detectar somente quando a população bacteriana está relativamente alta. Já a técnica de PCR que é uma das tecnicas consideradas mais sensíveis, não vem sendo utilizada na prática, pois esta alta sensibilidade não vem ocorrendo. Os protocolos não detectam as bacterioses no caso de infecção latente, onde o título bacteriano é relativamente baixo. Portanto, os demais trabalhos visaram aprimorar a detecção Xanthomonas albilineans desenvolvendo um protocolo LAMP, e comparando com outras tecnicas de detecção como isolamento em meio semi-seletivo, PCR e Nested-PCR, tanto para amostras sintomáticas quanto para assintomáticas. Para estas últimas onde a população do patógeno de maneira geral é baixa, aprimoramos um protocolo Nested para detectar escaldadura das folhas em infecções latentes, comparando técnicas de extração do fluído vascular combinado a quatro diferentes protocolos de extração de DNA.

The main focus of this work is the relationship between person and object. My initial hypothesis was that people are “creators of meaning”. It is one of our basic cognitive functions. We all need an understandable motivation to comprehend, consider and experience the everyday-life environment around us. We have all experienced a special connection with items that we own. This is also true for the subject of my analysis: luminaries. Light is the main protagonist of our life. Besides the obvious role in making us see the world around us, it has a major impact on modifying our feelings. Following deep literature research, I proposed to a selected group of volunteers a questionnaire to answer a concise but complex question: why we consider a special object that we care about. Despite this topic has been broadly analyzed in the field, it is my opinion that there is a lack of understanding about the influence that objects have on attachment bonds.  The purpose of my analysis was to understand what would make a person holding on to a lamp for all of his/her life and maybe giving it as an heirloom further on in the family. A variety of aspects related to the concept of bonding to a specific object have been identified: time, value to the person, love and care, irreplaceability, person-object interactions, an extension of self-identity, and emotions. In general, as a conclusion from my survey, I have identified three main aspects important for the person-luminaire bond:- the features of the person itself - the characteristic of a lamp in its duality - the relation bond between the two In general, people are focusing more broadly on the effect and the atmosphere created by the luminaire in the house and I can conclude that the connection is created through an important association between the psychological momentum and the home environment where they are immersed into. I finally proposed a guideline applicable to future design projects and to define the most important characteristics that an object should have to bond with a person for life.In future perspectives, an obvious implication would be to rethink our approach to design and drive professionals towards customer-oriented needs and expectations to extend the life cycle of products.

Waterborne or foodborne diseases are caused by consuming contaminated fluids or foods. The presence of pathogenic microorganisms can contaminate food or drinking water. These microorganisms can cause sickness even if they are present in minimal concentrations. The World Health Organization (WHO) has defined the standards for clean drinking water as the absence of E. coli in a 100 mL collected volume. Contaminated water or food can cause many diseases, and diarrhea is one of a prominent disease. Early detection of contamination in food or drinking water is critical. Conventional culture-based methods are time-consuming, labour intensive, and not suitable for on-site testing. Nucleic acid-based tests are sensitive and can rapidly detect pathogens. Microfluidic technology can play a significant role to develop low-cost, rapid, integrated, and portable nucleic acid-based detection devices. Microfluidic systems for isothermal amplification assays can be classified into two groups such as droplet-based and chamber-based systems. In this thesis, both droplet-based and chamber-based approaches were used to build the microfluidic methods for isothermal amplification assays. First, a simple electromechanical probe (tweezers) was developed that can manipulate a small aqueous droplet in a bi-layer oil phase. The tweezer consisted of two needles positioned close to each other and used polarization of the aqueous droplet in an applied electrical field to confine the droplet between the needles with minimal solid contact. AC electric potential was applied to the two metal electrodes. Droplet acquired a charge from the high voltage electrode and consequently performed an oscillatory motion with the same electrode. This droplet motion was controlled using two parameters of electric potential and frequency of the applied signal. Initially, electrically actuated droplet (0.3 µL) motion was investigated for a range of applied potential (400-960 Volts) and frequencies (0.1-1000 Hz). The droplet motion with high voltage electrode was characterized into three modes such as detachment, oscillation, and attachment. Mechanical motion of tweezer was used to transport droplet to various positions. Consequently, operations such as transportation, extraction, and merging were demonstrated. First, droplet (5 µL) transportation was characterized under the applied potential of 2000 Volts at various frequencies (5 to 1000 Hz). The droplet was successfully transported to the speed of 15 mm/s at higher frequencies (100 or 1000 Hz). Droplets of various volumes (12-80 µL) were extracted by increasing applied electric potential, from 0 to 6000 Volts at 5 Hz. Then, the operation of droplets merging was demonstrated using operational conditions for electrical tweezer. Finally, electrical tweezer was used to prepare samples for isothermal amplification assays. Two droplets consisted of various reagents of isothermal amplification assays, were transported and merged using the electrical tweezer. Then, a merged droplet (25 µL) was transported and immobilized in the amplification zone. The temperature of the amplification zone (~65°C) was maintained using an in-situ heater. DNA amplification was verified by measuring the off-chip end-point fluorescence intensity of isothermal assays. Second, an integrated microfluidic device has been developed to prepare a sample for isothermal amplification assays. And in-situ real-time amplification assays were performed to detect bacteria. The device consisted of two chambers (lysis and amplification) connected through a microchannel. A low-cost fabrication method was introduced to embed two resistive wire heaters around both chambers. Initially, bacteria cells were thermally lysed in the lysis chamber at 92°C for 5 min. Then, DNA was electrophoretically transported from lysis to the amplification chamber. The electric potential of 10 Volts was applied for 10 min for DNA transportation. Next, transported DNA was amplified at 65°C and DNA amplification was detected by measuring in-situ fluorescence intensity in the real-time format. The operation of the integrated microfluidic device was demonstrated in three steps. 1) Operation of individual components. 2) Operation of two components in a coupled format. 3) Integrated operation of three components with measurement of fluorescent intensity in a real-time format. The bacteria samples with the concentration of 100 CFU/mL were detected in less than one hour. / Thesis / Doctor of Philosophy (PhD)

The goal of this diploma’s thesis is to inform about present development of light sources, new technologies and about achieved parameters of these light sources. The thesis is mainly directed to describe efficiency of transformation electric energy to light at single types of light sources. There are described the concrete technical parameters of sources quoted by manufacturers and the contructional solution of single types of light sources. There is created a graphic comparation of electrotechnical and light parameters of the light sources.

Made available in DSpace on 2017-07-25T19:30:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-02-22 / Fundação Araucária de Apoio ao Desenvolvimento Científico e Tecnológico do Paraná / Sclerotinia sclerotiorum infect many plants, including crops of great economic importance as soybeans (Glycine max) and oilseed rape (Brassica sp.), responsible for great losses. The plant disease control is the most important practical objective of plant pathology, but the correct and rapid diagnosis are essential to define strategies for the diseases management. Molecular techniques are able to amplify specific fragments from small amounts of genetic material and powerful tools widely used in various areas, including the phytopathological diagnosis. Several techniques have been studied and designed for the amplification of nucleic acids, including the LAMP (Loop-mediated isothermal amplification), which has high specificity, sensitivity and is fast. The aim of this study was to develop and optimize a specific molecular test for S. sclerotiorum by LAMP, as well as its validation to oilseed pare and soybean seeds samples. A set of six primers was designed and evaluated for S. sclerotiorum sensitivity and specificity detection. The composition of the LAMP reaction was enhanced for real-time (SS-qLAMP) and direct analysis (SS-cLAMP). The DNA from 57 isolates of S. sclerotiorum, DNA from several other plant pathogens and DNA from different cultures was tested. The DNA of all isolates of S. sclerotiorum were detected but no the other DNA samples. When testing the limit of detection of reactions, a single copy detections was suggested. By SS-qLAMP two curves were generated which can be used to estimate the amount of mycelium and DNA of S. sclerotiorum present in the samples analyzed. Bothe developed tests (SS-qLAMP and SS-cLAMP) can be applied to several purposes, such as detection of the pathogen in plant, spore traps, soil and seeds samples. Using seed samples with different contamination level, the test was optimized for canola and soybean seeds, SS-qLAMP(Canola) and SS-qLAMP(Soybean), respectively, detecting the presence of the pathogen in samples up to 0.13% and 0.03% naturally contaminated for canola and soybean, respectively, and was able to detect contamination in samples not contaminated according incubation-based methods. The time require for the test was 4h and 30 minutes and 2 hours and 50 minutes for canola and soybeans, respectively, with no needs of large space for samples incubation, specialized analysts and able to analyzed many samples simultaneously. The proposed method SS-qLAMP was well-validated according the ISTA (International Seed Testing Association) rules to oilseed-rape and soybean seed samples. x / Sclerotinia sclerotiorum é um fungo que pode atacar diversas espécies vegetais, incluído culturas de grande importância econômica como soja (Glycine max) e canola (Brassica sp.), causando grandes prejuízos para as mesmas. O controle de doenças de plantas é o mais importante objetivo prático da Fitopatologia, mas, a correta e rápida diagnose da doença são pré-requisitos indispensáveis para definir as medidas para o manejo das mesmas. Técnicas moleculares capazes de amplificar fragmentos específicos a partir de pequenas quantidades de material genético são poderosas ferramentas amplamente utilizadas em diversas áreas, incluindo o diagnóstico fitopatológico. Diversas técnicas têm sido estudadas e criadas para a amplificação de ácidos nucleicos, entre elas a LAMP (Amplificação isotérmica mediada por “loops”), que apresenta alta especificidade, sensibilidade e é rápida. O objetivo do presente estudo foi desenvolver e otimizar um teste molecular específico para S. sclerotiorum a base de LAMP, com obtenção de resultados em tempo real e quantitativos (qLAMP) e análise visual direta de resultados (cLAMP), assim como a validação do mesmo para ser utilizado na análise de amostras de sementes de canola e soja. Um conjunto de seis primers foi desenhado e avaliado quanto a sensibilidade e especificidade de detecção de S. sclerotiorum. A composição da reação de LAMP foi otimizada quanto a concentração de diversos componentes tanto para a análise em tempo real como direta, compondo, respectivamente as reações de SS-qLAMP e SS-cLAMP, conforme necessário. Testou-se o DNA de 57 isolados de S. sclerotiorum, o DNA de diversos outros fitopatógenos e o DNA de diversas culturas. O DNA de todos os isolados de S. sclerotiorum foram detectados mas não o de outros fitopatógenos e de plantas. Ao testar o limite de detecção das reações, não houve um limite de detecção, sugerindo que a presença de qualquer molécula de DNA alvo seria detectada. Pelo método quantitativo foi possível gerar duas curvas pelas quais pode-se estimar a quantidade de micélio e de DNA de S. sclerotiorum presente na amostra analisada. Desenvolveu-se um teste específico para S. sclerotiorum a base de LAMP, denominada SS-LAMP que pode ser aplicado em diversos casos, como a detecção do patógeno em amostras de plantas, armadilhas de esporos, de solo e sementes. Otimizou-se o teste SS-LAMP para sementes de canola e soja, SS-qLAMP(Canola) e SS-qLAMP(Soja), respectivamente, detectando-se a presença do patógeno em amostras com até 0,13% e 0,03% de contaminação natural, para canola e soja, respectivamente, além da detecção em amostras que os métodos tradicionais não detectaram. O tempo total do método foi de 4h e 30 minutos para canola e 2h e 50 minutos para soja, sem a necessidade de amplo espaço para incubação das amostras, pessoal especializado para análise e com a possibilidade de diversas amostras serem analisadas concomitantemente. Assim validou-se o método proposto SS-qLAMP(Soja) segundo as regras da ISTA (International Seed Testing Association – Associação Internacional de Análise de Sementes).

Made available in DSpace on 2016-08-17T18:39:41Z (GMT). No. of bitstreams: 1 4098.pdf: 4555828 bytes, checksum: eeec91702a4d283144c291e30559f09c (MD5) Previous issue date: 2011-02-18 / In this paper we proposed and analyzed methodologies using the technology of Cell-Free Fetal Nucleic Acid-Free (cffDNA = Cell Free Fetal DNA) in noninvasive prenatal diagnosis (NIPD). Due to the modern technologies employed and their repercussion among the involved families, we sought to discuss some ethical, social and legal implications. Contrary to the popular belief that the placenta forms an impermeable barrier between mother and child, there is bidirectional traffic between the fetus and mother during pregnancy. Several studies have shown that not only intact cells but also fetal cell-free fetal nucleic acids (cffNA, ie, DNA and RNA) cross the placenta and travels in the mother's bloodstream. Four different applications of analysis technology were identified: cffDNA: a) Prenatal Sex Determination, used in pregnancies under the risk of sexual transmitted diseases and performed through the detection of the Y chromosome b) The diagnosis of certain diseases of a single gene through the detection of paternally inherited mutation c) Fetal Aneuploidy, such as Down syndrome, where chromosomal abnormalities may occur d) identifying the type of fetal blood in pregnancies under the risk of incompatibility, especially RhD. To carry out this work it was used the pre-natal determination of sex in 53 pregnant women at different gestational periods. For that it was proposed and tested methods of DNA extraction, amplification of DNA obtained by PCR (Polymerase Chain Reaction) and a new methodology called LAMP (Loop Mediated Isothermal Amplification) and the analysis of final results. Furthermore, the efficiency of LAMP and PCR was compared by amplifying different segments of the Y chromosome: DSY14 and TSPY. In three cases the samples were discarded because there was no fetal sex confirmation after molecular tests due to loss of contact with pregnant women. In two other cases the results pointed to male fetuses whilst the ultrasound confirmed these fetuses as females due to contamination. Finally it was obtained 28 male samples (58.33%) with amplification of the sequences of the Y chromosome and 20 female samples (41.67%) that did not amplify the sequence of the Y chromosome but only for the control. These results showed that the amplification lamp is more efficient than PCR, in the analysis of DSY14, the limit of detection is 10 pg and 0.1 pg for LAMP and PCR respectively. It was concluded that the amplification using the LAMP method is faster (60 min) and has a high sensitivity and specificity and does not require sophisticated equipment for reaction if compared to the PCR method. These characteristics make this methodology feaseable in laboratories with limited resources. / Neste trabalho propusemos e analisamos metodologias empregadas no uso da tecnologia dos Ácidos Nucléicos Livres de Células Fetais (cffDNA = Cell Free Fetal DNA) no diagnóstico pré-natal não invasivo (DPIN = Non-invasive Prenatal Diagnosis). Dada a modernidade das tecnologias empregadas e a sua repercussão nas famílias envolvidas, procuramos discutir algumas implicações éticas, sociais e legais. Ao contrário da crença popular de que a placenta constitui uma barreira impermeável entre mãe e filho, há tráfego bidirecional entre o feto e a mãe durante a gravidez. Vários estudos têm demonstrado que tanto células fetais intactas e ácidos nucléicos livres de células fetais (cffNA, ou seja, DNA e RNA) atravessam a placenta e circulam na corrente sanguínea materna. Identificamos quatro diferentes aplicações da tecnologia de análise do cffDNA: a) Determinação pré-natal do sexo, utilizada em gestações com risco de uma doença ligada ao sexo e realizada através da detecção do cromossomo masculino Y; b) O diagnóstico de certas doenças de gene único, normalmente através da detecção de mutação herdada paternalmente; c) Aneuploidia fetal, tal como a Síndrome de Down, onde há alterações cromossômicas; d) Detecção do tipo de sangue fetal em gestações com risco de incompatibilidade, sobretudo RhD. Para realizar este trabalho utilizamos a determinação pré-natal do sexo em 53 gestantes em diferentes períodos gestacionais. Para tal, testamos e propusemos metodologias de extração do DNA; amplificação do DNA extraído através da técnica da PCR (Polymerase Chain Reaction) e de uma nova metodologia denominada LAMP (Loop Mediated Isothermal Amplification); e a análise final dos resultados. Além disso, comparamos a eficiência da PCR e LAMP amplificando diferentes segmentos do cromossomo Y: DSY14 e TSPY. Em três casos as amostras foram descartadas porque não houve a confirmação do sexo fetal após os testes moleculares devido à perda de contato com as gestantes. Em outros dois casos os resultados apontaram para fetos masculinos, entretanto, no ultra-som eles foram confirmados como femininos, provavelmente devido a uma contaminção. No final obtivemos 28 amostras (58,33%) masculinas, com a amplificação das sequências do cromossomo Y e 20 amostras (41,67%) femininas que não amplificaram a sequência do cromossomo Y e apenas para o controle. Nossos resultados demonstraram que a amplificação por LAMP é mais eficiente que a PCR na análise de DSY14, sendo que o limite de detecção é de 10 pg e 0,1 pg, para a PCR e o LAMP, respectivamente. Concluímos que a amplificação utilizando a metodologia de LAMP é mais rápida (60 minutos) e apresenta uma alta especificidade, sensibilidade e não requer equipamentos sofisticados para reação, comparada com a técnica da PCR. Tais características viabilizam esta metodologia em laboratórios com poucos recursos.