Дисертації з теми "Molecular Diagnosi"
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D'AVERSA, Elisabetta. "Innovative approaches for molecular diagnosis of genetic diseases." Doctoral thesis, Università degli studi di Ferrara, 2019. http://hdl.handle.net/11392/2488091.
Повний текст джерелаLa diagnosi di patologie genetiche fetali viene effettuata a partire da materiale biologico prelevato tramite tecniche invasive, le quali presentano un elevato rischio di aborto. La diagnosi prenatale non invasiva, effettuata a partire da ccffDNA (DNA circolante fetale), isolato da sangue materno, negli ultimi anni, ha avuto una crescita esponenziale. La diagnosi del sesso, in età prenatale, permette di definire il rischio di malattie legate al cromosoma X. La prima parte della tesi ha come obbiettivo, quindi, lo sviluppo di approcci diagnostici molecolari non invasivi, rapidi e sensibili per la determinazione del sesso fetale, in particolare a settimane di gestazione precoci. Per verificare la presenza del gene SRY, sul cromosoma Y, nel ccffDNA, estratto da plasma materno, è stata impiegata, in primis, la Real Time PCR. I risultati hanno confermato il limite di rilevabilità della tecnica (9 settimane) individuato in uno studio preliminare del gruppo di ricerca. Per settimane di gestazione precoci è stata impiegata la tecnologia innovativa digital droplet PCR (ddPCR): dopo aver ottimizzato le condizioni sperimentali, sono stati analizzati campioni a settimane di gestazione comprese fra le 12 e le 4.5, e per tutti la tecnica si è dimostrata accurata al 100% nel determinare correttamente il sesso fetale. La β talassemia è una patologia autosomica recessiva associata all’assenza o riduzione delle catene β globiniche dell’emoglobina. I soggetti che presentano la forma più severa della patologia non possono sopravvivere senza trasfusioni associate a ferrochelanti. L’unico vero trattamento definitivo è il trapianto di midollo, il quale però è associato a forti rischi. Sono stati studiati, quindi, approcci terapeutici innovativi con l’obbiettivo di una terapia personalizzata. L’identificazione della specifica alterazione molecolare è, quindi, di fondamentale importanza al fine di scegliere la corretta strategia terapeutica. La seconda parte della ricerca ha, quindi, come obbiettivo lo sviluppo di approcci molecolari diagnostici pre- e postnatali rapidi, sensibili e poco costosi per l’identificazione delle più frequenti mutazioni talassemiche nel Mediterraneo (β039, β+IVSI-110, β+IVSI-6, β0IVSI-1). Per la diagnosi postnatale, è stata impiegata, in primis, la tecnologia BiacoreTM: una sonda mutata e normale sono state immobilizzate sul chip, validate e, successivamente, è stato estratto il DNA genomico di soggetti sani e pazienti omozigoti ed eterozigoti per le mutazioni considerate, ed effettuata l’analisi. Per tutti i campioni analizzati è stato possibile discriminare il diverso genotipo. Un secondo approccio ha previsto il disegno di saggi di genotipizzazione da utilizzare in Real Time PCR. Ottimizzati e validati, i saggi hanno permesso una corretta diagnosi di tutti i pazienti considerati. Lo stesso approccio, basato sui saggi di genotipizzazione, è stato utilizzato per la diagnosi prenatale delle mutazioni ereditate per via paterna. I campioni di ccffDNA sono stati preamplificati e analizzati, mostrando come questo approccio sia stato in grado di identificare il genotipo fetale a partire dalla nona settimana di gestazione. Per poter estendere la diagnosi prenatale a settimane di gestazione precoci e a mutazioni ereditate sia per via materna che da entrambi i genitori è stata utilizzata la ddPCR come approccio molecolare. I saggi di genotipizzazione per le mutazioni β039 e β+IVSI-110 sono stati ottimizzati, validati e utilizzati per l’analisi del ccffDNA. Per tutti i campioni in cui la mutazione è ereditata per via paterna, il genotipo fetale è stato correttamente individuato fino alla quinta settimana di gestazione. Per i campioni in cui solo la madre o entrambi i genitori sono portatori, è stato possibile individuare due intervalli, sulla base del rapporto fra allele mutato e normale, statisticamente separati, necessari per considerare il feto non portatore o eterozigote.
Soler, Aznar Maria. "Nanoplasmonic biosensors for clinical diagnosis at the point of care." Doctoral thesis, Universitat Autònoma de Barcelona, 2015. http://hdl.handle.net/10803/298172.
Повний текст джерелаThis Doctoral Thesis focuses on the development of novel analytical methodologies in optical biosensors as alternative solutions for diagnosis or therapy monitoring of relevant diseases, such as allergy, celiac disease or cancer. In particular, we propose the use of nanoplasmonic biosensors for a rapid, sensitive and label-free detection of biomarkers present in human fluids. Both the well-known Surface Plasmon Resonance (SPR) biosensor and an innovative nanoplasmonic biosensor based on gold nanodisks surfaces have been evaluated for their real application in the clinical field. The different biosensor methodologies make use of antibodies, either as biorecognition elements in immunoassays or as specific disease biomarkers for diagnostics. First, an in-depth study of two site-directed antibody immobilization strategies is presented for the direct immunoassay of protein biomarkers in biological fluids. In second place, a novel immunosensing strategy is proposed for the detection of gluten-derivative peptides in urine as a rapid and non-invasive technique for dietary control in celiac patients. On the other hand, two assays have been developed employing the nanoplasmonic biosensor to detect blood circulating antibodies as disease biomarkers. First, we have designed an alternative approach for drug allergy diagnosis (in particular for amoxicillin) based on dendrimer-based receptors, which enable the detection IgE antibodies directly in serum. And second, a new biosensing strategy is assessed to quantify specific tumor-related autoantibodies for the early diagnosis of colorectal cancer. The work in this Thesis combines the wide knowledge of the research group in the design and fabrication of powerful biosensor technology with the development of surface activation chemistry and bioanalytical techniques to overcome current challenges related to costly and time-consuming clinical analysis. Besides, the strong experience of our research group in technological transfer and the established collaborations during this doctoral work with companies as Biomedal S.L. or Protein Alternatives S.L. open up interesting opportunities to facilitate the technology-transfer process for the real implementation of Point-of-Care biosensors.
GRIONI, ANDREA. "Application of modern data science to genomics and clinical research." Doctoral thesis, Università degli Studi di Milano-Bicocca, 2020. http://hdl.handle.net/10281/279991.
Повний текст джерелаAfter the completion of the human genome project in April 2003, the continuous flow of sequencing data and the development of new databases began to transform the field of genomics into data-driven science. Bioinformatics analyses raw experimental data with the aim to obtain information describing biological processes, thus providing a powerful tool to investigate specific molecular and genetic mechanisms. This domain knowledge in combination with genomics allows to decipher the interrelationships between genes, regulatory elements, metabolic pathways, and protein interactions. Deep learning, a subdiscipline of machine learning, has been recently applied to the field of genomics, leading to remarkable results. The two main objectives of this study were: the development and application of bioinformatic tools for the study of the genetic basis of acute lymphoblastic leukaemia, and the usage of deep learning techniques for the identification of small non-coding RNA elements in the human genome. This dissertation provides a comprehensive overview of the recent evolution of genomics as an interdisciplinary field of research strongly associated with computer science and data analysis.
Shirahata, Mitsuaki. "Gene Expression-Based Molecular Diagnostic System for Malignant Gliomas Is Superior to Histological Diagnosis." Kyoto University, 2008. http://hdl.handle.net/2433/124241.
Повний текст джерелаRamiro, Juliana. "Detecção molecular de fungos fitopatogênicos associados às sementes de soja." Universidade de São Paulo, 2015. http://www.teses.usp.br/teses/disponiveis/11/11135/tde-28042015-143631/.
Повний текст джерелаSeed-born pathogenic fungi in soybean can cause serious damage to the crop, as well as direct damage by reducing seeds germination, vigor and emergence. The detection and accurate identification of plant pathogens is one of the most important strategies to initiate preventive and curative measures in the management of plant diseases. Particular attention should be taken in the detection of pathogens in seeds in order to avoid introduction and spread of pathogens in areas where they do not occur. The traditionally used methods for detection and identification of seed-born pathogenic fungi are often time consuming, laborious and require extensive knowledge of classical taxonomy. Molecular methods have been used to detect, identify and quantify a long list of plant pathogenic fungi. Quantitative real time polymerase chain reaction (qPCR) is currently considered the most efficient technique for the detection of pathogens because it does not require specialized taxonomical knowledge to interpret its results. Given the importance and implications of rapid and successful diagnosis of seed-born pathogenic fungi in soybean, this study aimed to establish a qPCR methodology to increase the detection efficiency of plant pathogenic fungi Sclerotinia sclerotiorum, Colletotrichum truncatum, Phomopsis spp. and Corynespora cassiicola, the most commonly occurring seed-born pathogenic fungi. Primers and TaqMan® hydrolysis probes were designed for these four pathogens and tested for specificity and sensitivity. Standard curves were established to quantify these pathogens, based on serial dilutions of the target DNA and specific primers and probes. Samples of naturally infected soybean seed from the states of Goiás, Minas Gerais and Paraná were subjected to detection tests using qPCR and traditional methods, for comparison purposes. From all primers and probes designed, only those for Phomopsis spp. and S. sclerotiorum showed up specificity and sensitivity, enabling their use to detect of these pathogens. Detection by traditional tests, resulted in a maximum Phomopsis spp. incidence of 2.75% in a sample from Minas Gerais and S. sclerotiorum was not detected in any of the samples. The detection and quantification of these pathogens by qPCR revealed the presence of Phomopsis spp. in all tested samples, the highest incidence level of 6.75% in a sample from Minas Gerais. S. sclerotiorum was not detected in any sample assessed by qPCR method. In comparison with traditional methods, qPCR was more sensitive in detecting Phomopsis spp. in soybean seeds.
Wang, Lei. "Molecular Probes for Pancreatic Cancer Imaging." PDXScholar, 2016. http://pdxscholar.library.pdx.edu/open_access_etds/3108.
Повний текст джерелаCATARINO, Aricléia de Moraes. "Detecção de vírus da videira por RT-PCR em tempo real e por extensão de primers alelo-específicos e caracterização molecular de isolados do Nordeste Brasileiro." Universidade Federal Rural de Pernambuco, 2015. http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/5995.
Повний текст джерелаMade available in DSpace on 2016-11-28T13:27:09Z (GMT). No. of bitstreams: 1 Aricleia de Moraes Catarino.pdf: 1031221 bytes, checksum: 5bd0ffb507daa4b9b04ebd494fd2269c (MD5) Previous issue date: 2015-02-27
The grapevine (Vitis spp.) belongs to the family of Vitaceae, being the botanical species V. vinifera L. and V. labrusca L. the most and widely cultivated, due to their products consumed as fresh fruits, jam, juices and wines. Despite the high economical importance, several factors may severely affect this crop, including the diseases caused by viruses. This study aimed to verify the incidence of virus present in commercial vineyards of two producing areas in Northeastern Brazil and evaluate the efficiency of some molecular methods for detecting and identifying viral species associated with grapevine. Materials showing or not symptoms were collected from grapevine genotypes in vineyards of Pernambuco, Paraiba, Bahia and Rio Grande do Sul, Brazil, and Locorotondo, Province of Bari, of the Puglia Region, Italy. The first part of the work was conducted at the Virology Laboratory of the Embrapa Uva e Vinho, RS, Brazil. For the identification of viral agents in the samples collected in Brazil, the extraction of total RNA was performed, cDNAs were obtained and tested by real time RT-PCR, using primers and probes specific for the following viruses: Grapevine rupestris stem pitting-associated virus (GRSPaV), Grapevine virus A (GVA), Grapevine virus B (GVB), Grapevine leafroll-associated virus 2, 3 e 4 (GLRaV-2, -3 e -4), Grapevine fleck virus (GFkV), Grapevine rupestris vein feathering virus (GRVFV) and Grapevine fanleaf virus (GFLV). DNA fragments, products of the RT-qPCR, corresponding to the CP gene of each virus were eluted, linked to pGEM-T Easy vector (Promega) and used to transform bacteria. The plasmid DNA was extracted from transformed bacterial colonies, confirming the presence of the cloned fragments, which were sequenced. The grapevine material collected in Locorotondo was processed at the Istituto per la Protezione Sostenibile delle Piante (CNR-IPSP) and at the Dipartimento di Scienze del Suolo, della Pianta e degli Alimenti, Università degli Studi “Aldo Moro”. In order to detect in multiplex test the most relevant viruses involved in the aetiology of fanleaf degeneration and the complexes of leafroll and rugose wood of grapevine, amplification techniques based on Allele Specific Primer Extension (ASPE) were tested by using the obtained cDNAs. The results showed that the techniques aggregate some advantages, such as reduction in time and relative simplicity of implementation, completely eliminating the use of toxic reagents, such as the ethidium bromide. The use of multiplex facilitates amplification of multiple targets in a single reaction, reducing the time and cost of the analyzes.
A videira (Vitis spp.) pertence à família Vitaceae, sendo as espécies botânicas V. vinifera L. e V. labrusca L. cultivadas em maior escala devido seus produtos, consumidos na forma de frutos in natura, geleias, sucos e vinhos. Apesar da grande importância econômica, vários fatores podem comprometer a produção desta cultura, incluindo as doenças causadas por vírus. O presente trabalho teve como objetivos verificar a incidência de vírus presentes em vinhedos comerciais de duas áreas produtoras do Nordeste do Brasil e avaliar a eficiência de alguns métodos moleculares para detecção e identificação de espécies virais associadas à videira. Amostras, apresentando ou não sintomas, foram coletados de genótipos de videira em propriedades situadas em Pernambuco, Paraíba, Bahia e Rio Grande do Sul, Brasil, e em Locorotondo, Província de Bari, Região da Puglia, Itália. A primeira parte do trabalho foi conduzida no Laboratório de Virologia da Embrapa Uva e Vinho, RS, Brasil. Visando a identificação dos agentes virais, nas amostras coletadas no Brasil, foram realizadas as extrações do RNA total, obtidos os cDNAs e testados por PCR em Tempo Real, empregando-se primers e sondas, específicos para os seguintes vírus: Grapevine rupestris stem pitting-associated virus (GRSPaV), Grapevine virus A (GVA), Grapevine virus B (GVB), Grapevine leafroll-associated virus 2, 3 e 4 (GLRaV-2, -3 e -4), Grapevine fleck virus (GFkV), Grapevine rupestris vein feathering virus (GRVFV) e Grapevine fanleaf virus (GFLV). Fragmentos de DNA, produtos da RTq-PCR, correspondentes ao gene da CP de cada vírus foram eluídos, ligados ao vetor pGEM-T Easy (Promega) e utilizados na transformação de bactéria. Foi extraído o DNA plasmidial das colônias bacterianas transformadas, confirmando-se a presença dos fragmentos clonados, os quais foram sequenciados. A segunda parte, realizada com o material coletado em Locorotondo, foi processada no Istituto per la Protezione Sostenibile delle Piante (CNR-IPSP) e no Dipartimento di Scienze del Suolo, della Pianta e degli Alimenti, Università degli Studi “Aldo Moro”. Foram utilizadas, a partir de cDNAs obtidos, técnicas de amplificação baseadas na Allele Specific Primer Extension (ASPE), visando detectar em teste multiplex os vírus mais relevantes envolvidos na etiologia da degenerescência e nos complexos do enrolamento das folhas e do lenho rugoso da videira. Os resultados obtidos mostraram que as técnicas agregam algumas vantagens, como a redução no tempo e relativa simplicidade de execução, eliminando completamente o uso de reagentes tóxicos, a exemplo do brometo de etídeo. O uso de multiplex facilita a amplificação de múltiplos alvos em uma única reação, reduzindo o tempo e o custo das análises.
顔鴻儀 and Hung-yee Ngan. "Molecular diagnosis of penicilliosis marneffei." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2001. http://hub.hku.hk/bib/B31970035.
Повний текст джерелаYau, Shu Ching. "Molecular diagnosis of neuromuscular disorders." Thesis, King's College London (University of London), 2003. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.402033.
Повний текст джерелаNgan, Hung-yee. "Molecular diagnosis of penicilliosis marneffei." Hong Kong : University of Hong Kong, 2001. http://sunzi.lib.hku.hk/hkuto/record.jsp?B23595978.
Повний текст джерелаCheung, Pik-shan, and 張碧珊. "Molecular diagnosis of soft tissue tumours." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2009. http://hub.hku.hk/bib/B42905370.
Повний текст джерелаCheung, Pik-shan. "Molecular diagnosis of soft tissue tumours." Click to view the E-thesis via HKUTO, 2009. http://sunzi.lib.hku.hk/hkuto/record/B42905370.
Повний текст джерелаGorton, Rebecca Louise. "Molecular diagnosis of invasive fungal disease." Thesis, University College London (University of London), 2018. http://discovery.ucl.ac.uk/10047377/.
Повний текст джерелаZetsma, Julia Wendelina. "Molecular diagnosis and epidemiology of Mycoplasma pneumoniae." [S.l. : Amsterdam : s.n.] ; Universiteit van Amsterdam [Host], 2000. http://dare.uva.nl/document/83311.
Повний текст джерелаEaton, Michael Campbell. "Assessment of CD44 and K19 as markers for circulating breast cancer cells using immunobead RT-PCR /." Title page, table of contents and abstract only, 1997. http://web4.library.adelaide.edu.au/theses/09MD/09mde14.pdf.
Повний текст джерелаForsgren, Eva. "Molecular diagnosis and characterization of honey bee pathogens /." Uppsala : Department of Ecology, Swedish University of Agricultural Sciences, 2009. http://epsilon.slu.se/200979.pdf.
Повний текст джерелаHemmings, Lance. "The molecular diagnosis of Echinococcus multilocularis in man." Thesis, Imperial College London, 1989. http://hdl.handle.net/10044/1/47473.
Повний текст джерелаDhorda, Mehul. "Molecular parasitology and diagnosis of Malaria in pregnancy." Paris 6, 2010. http://www.theses.fr/2010PA066405.
Повний текст джерелаLoukola, Anu-Maria. "Molecular diagnosis of hereditary nonpolyposis colorectal cancer (HNPCC)." Helsinki : University of Helsinki, 2000. http://ethesis.helsinki.fi/julkaisut/laa/haart/vk/loukola/.
Повний текст джерелаSchwarz, Emanuel. "Molecular diagnostic aids for neuropsychiatric disorders." Thesis, University of Cambridge, 2009. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.611585.
Повний текст джерелаSiu, Kit-hang, and 蕭傑恆. "Molecular characterization of multi-drug resistance mechanisms in mycobacterium tuberculosis." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2010. http://hub.hku.hk/bib/B46076219.
Повний текст джерелаMérola, Claudia Braida. "Molecular analysis of myotonic dystrophy type 1 patients with an unusual molecular diagnosis." Thesis, University of Glasgow, 2008. http://theses.gla.ac.uk/359/.
Повний текст джерелаBraida, Claudia. "Molecular analysis of myotonic dystrophy type 1 patients with an unusual molecular diagnosis." Thesis restricted. Connect to e-thesis to view abstract, 2008. http://theses.gla.ac.uk/359/.
Повний текст джерелаPh.D. thesis submitted to the Division of Molecular Genetics, Institute of Biomedical and Life Sciences, University of Glasgow, 2008. Includes bibliographical references. Print version also available.
Elliott, Suzanne Louise. "Molecular analysis as an adjunct in the diagnosis and management of leukaemia." Thesis, Queensland University of Technology, 1992.
Знайти повний текст джерелаSaleh, Meriam Naim. "Detecting Giardia: Clinical and Molecular Identification." Diss., Virginia Tech, 2017. http://hdl.handle.net/10919/89367.
Повний текст джерелаPHD
Zurkiya, Omar. "Magnetic Resonance Molecular Imaging Using Iron Oxide Nanoparticles." Diss., Georgia Institute of Technology, 2006. http://hdl.handle.net/1853/19848.
Повний текст джерелаYip, Kam-tong, and 葉錦棠. "Comparative study on molecular and serological diagnosis of tuberculosis." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2000. http://hub.hku.hk/bib/B31969902.
Повний текст джерелаAl-Jawabreh, Amer. "Molecular Epidemiology, Clinical Molecular Diagnosis and Genetic Diversity of Cutaneous Leishmaniasis in Jericho, Palestine." Doctoral thesis, Humboldt-Universität zu Berlin, Medizinische Fakultät - Universitätsklinikum Charité, 2006. http://dx.doi.org/10.18452/15391.
Повний текст джерелаIn this study we compared the sensitivity of the diagnosis of Giemsa-stained skin scrapings by standardized graded direct microscopy with that of ITS1-PCR. ITS1-PCR showed a sensitivity of 87% with positive predictive value of 100% and a specificity of 100% with negative predictive value of 85%. In-vitro cultivation using NNN medium and direct smear microscopy of Giemsa-stained slides, PCR amplifying region 1 of internal transcribed spacer (ITS1) using skin scrapings spotted on filter papers (FP) and unstained tissue smears (US) were compared. PCR using US was more sensitive than all other methods Molecular epidemiology was used to study the distribution of Leishmania species in Jericho. Spatial analysis showed three statistically significant clusters of CL, one cluster for L. major and two clusters for L. tropica. In the case of space-time, four clusters for CL, two for L. major and three for L. tropica were detected. A total of 106 strains isolated in different endemic regions of Central Asia, Middle East and Africa were analysed using 10 pairs of microsatellite markers under two cluster methods: distance and model-based. Markers were designed to amplify microsatellite loci identified in the genome sequence of L. major on chromosomes 1, 3, 5, 21 and 35. Seven discrete populations of L. major including two genetically isolated populations in the Middle East were revealed.
Dawson, Sarah-Jane. "Molecular biomarkers in breast cancer." Thesis, University of Cambridge, 2011. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.609742.
Повний текст джерелаYip, Kam-tong. "Comparative study on molecular and serological diagnosis of tuberculosis." Click to view the E-thesis via HKUTO, 2000. http://sunzi.lib.hku.hk/hkuto/record/B31969902.
Повний текст джерелаau, N. Phillips@murdoch edu, and Nyree Phillips. "Diagnosis, molecular epidemiology and control of avian intestinal spirochaetosis." Murdoch University, 2006. http://wwwlib.murdoch.edu.au/adt/browse/view/adt-MU20070129.142632.
Повний текст джерелаPhillips, Nyree Dale. "Diagnosis, molecular epidemiology and control of avian intestinal spirochaetosis /." Access via Murdoch University Digital Theses Project, 2006. http://wwwlib.murdoch.edu.au/adt/browse/view/adt-MU20070129.142632.
Повний текст джерелаKwong, Tsz-ching, and 鄺芷晴. "The role of molecular diagnosis of drug resistant tuberculosis." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2015. http://hdl.handle.net/10722/208588.
Повний текст джерелаpublished_or_final_version
Microbiology
Master
Master of Philosophy
Phillips, Nyree. "Diagnosis, molecular epidemiology and control of avian intestinal spirochaetosis." Thesis, Phillips, Nyree (2006) Diagnosis, molecular epidemiology and control of avian intestinal spirochaetosis. PhD thesis, Murdoch University, 2006. https://researchrepository.murdoch.edu.au/id/eprint/259/.
Повний текст джерелаPhillips, Nyree. "Diagnosis, molecular epidemiology and control of avian intestinal spirochaetosis." Phillips, Nyree (2006) Diagnosis, molecular epidemiology and control of avian intestinal spirochaetosis. PhD thesis, Murdoch University, 2006. http://researchrepository.murdoch.edu.au/259/.
Повний текст джерелаCheung, Nga-yin Annie, and 張雅賢. "Cervical cancer screening: evolution from Paptest to molecular markers." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2011. http://hub.hku.hk/bib/B46540465.
Повний текст джерелаTsang, Lai-ying, and 曾麗凝. "Rapid detection of mycobacterium tuberculosis using single-tube nestedreal time PCR." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2010. http://hub.hku.hk/bib/B44670680.
Повний текст джерелаVarawalla, Nermeen Y. "Molecular genetics of beta thalassaemia in Asian Indians : basis for prenatal diagnosis." Thesis, University of Oxford, 1992. http://ora.ox.ac.uk/objects/uuid:f3a2a0a7-3d14-4dcf-a6fc-616db75119bf.
Повний текст джерелаNascimento, Aline Kelly Queiroz do. "Technological innovations for diagnosis of plant viruses and characterization from biotypes of cowpea aphid-borne mosaic virus." Universidade Federal do CearÃ, 2014. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=13205.
Повний текст джерелаPlant virus identification and characterization can be achived by several methods based in the biological, morphological, cytological, serological and molecular virus properties. The molecular properties have been used with frequency for vÃrus identification and characterization and the reverse transcription polymerase chain reaction (RT-PCR) has constituted an efficient and precise method for researches with RNA plant viruses. On the other hand, the enzyme-linked immunosorbent assay (ELISA) is the most used serological method for virus detection in plant tissues. A techncal innovation developed for plant virus identification represents a great technological development and support for plant virus research. A new approach involving virus particle immune precipitation to be used for RNA amplification by RT-PCR named IP-RT-PCR was sucessefuly used for amplification of RNA fragments from five virus species from the genera Comovirus, Cucumovirus, Potyvirus and Sobemovirus. Considering that the immune biological Companies have developed several DAS-ELISA kits, but neither of them produce and commercialize PTA-ELISA kits, a simple and practical PTA-ELISA kit was developed and validated for plant virus detection. The second part of the research had the objective to study, comparatively, the biological, serological and molecular properties of four plant virus isolates obtained from naturally infected Passiflora edulis (PWV-PET and PWV-GUA) and from naturally infected Vigna unguiculata (CABMV-BV and CABMV-FOR) with the objective to elucidate the identity of the causal agent of passion fruit woodiness in Brazil. In host range studies onle Canavalia ensiformes and Macroptilium lathyroides were infected by virus isolates obtained from cowpea and from passion fruit. The isolate PWV-GUA was purified from systemically infected M. lathyroides plants and the virus purified preparation (18.24 mg of virus.ml-1) was used for rabitt immunization for polyclonal antiserum production, which showed a title of 1:128,000 in PTA-ELISA. The electrophoresis analysis of the purified virus showed a unique capsidial protein with 34 kDA. Plant virus interaction studies in C. ensiformis indicated unilateral cross protection between PWV-GUA and CABMV-FOR. On the other hand, the isolate PWV-PET did not cross protect passion fruit plants against PWV-GUA. Filogenetic analysis of nucleotiode sequencies from cDNA fragments corresponding to coat protein (CP) genes amplified by IP-RT-PCR from the genomic virus isolates compared with virus sequencies from the Genbank grouped according to the host specifities. Based on the biological, serological and mainly molecular results, the virus isolates studied were classified into two biotypes: Biotype CABMC-C (Cowpea) to include isolates obtained from cowpea that do not infect passion fruit, and biotype CABMV-P (Passion fruit) to include the virus isolates responsible for the passion fruit woodiness in Brazil.
A identificaÃÃo e a caracterizaÃÃo de vÃrus de planta podem ser realizadas por vÃrios mÃtodos envolvendo propriedades morfolÃgicas, biolÃgicas, citolÃgicas, moleculares e sorolÃgicas. As tÃcnicas moleculares tÃm sido usadas com frequencia para identificaÃÃo e caracterizaÃÃo de vÃrus, e a tÃcnica de âreverse transcription polymerase chain reactionâ (RT-PCR) tem se constituÃdo em mÃtodo eficiente e preciso para pesquisas com vÃrus de planta com genoma de RNA. De outra parte, a tÃcnica de enzyme-linked immunosorbent assay (ELISA) constitui o mÃtodo sorolÃgico mais usado para detecÃÃo de vÃrus em tecidos vegetais. Uma inovaÃÃo tecnolÃgica desenvolvida nesta pesquisa para diagnose de vÃrus de planta representa grande avanÃo tecnolÃgico e suporte para pesquisa em virologia vegetal. A inovaÃÃo envolvendo a imunoprecipitaÃÃo (IP) de partÃculas de vÃrus para uso na RT-PCR denominada de IP-RT-PCR foi usada com sucesso para amplificaÃÃo de fragmentos de RNA de cinco espÃcies de vÃrus dos gÃneros Comovirus, Cucumovirus, Potyvirus e Sobemovirus. Considerando que kits de DAS-ELISA tÃm sido produzidos e comercializados por companhias de imunobiologicos, mas nenhuma companhia produz kits de PTA-ELISA, um kit simples e prÃtico de PTA-ELISA foi desenvolvido e validado para detecÃÃo de vÃrus de planta. A segunda etapa da pesquisa teve como objetivo estudar as propriedades biolÃgicas, sorolÃgicas e moleculares de isolados de vÃrus do gÃnero Potyvirus obtidos de maracujazeiro (Passiflora edulis) (PWV-PET e PWV-GUA) e isolados de Cowpea aphid-borne mosaic virus (CABMV-FOR e CABMV-BV) obtidos de feijoeiro caupi (Vigna unguiculata), visando elucidar a identidade do agente causal do endurecimento dos frutos do maracujazeiro no Brasil. Em estudos de gama de plantas hospedeiras, somente Canavalia ensiformis e Macroptilium lathyroides foram infetadas por isolados obtidos de maracujazeiro e de feijoeiro caupi. O PWV-GUA foi purificado a partir de plantas de M. lathyroides sistemicamente infetadas e a preparaÃÃo viral purificada (18,24 mg de vÃrus.ml-1) foi usada para imunizaÃÃo de coelho com a produÃÃo de antissoro policlonal com tÃtulo de 1:128.000 em PTA-ELISA. AnÃlise eletroforÃtica da preparaÃÃo viral purificada revelou uma Ãnica proteÃna capisidial com peso molecular de 34 kDa. Experimentos de interaÃÃo entre os isolados virais em C. ensiformis indicaram proteÃÃo unilateral entre PWV-GUA e CABMV-FOR. De outra parte, o isolado PWV-PET nÃo protegeu plantas de maracujazeiro contra a super infecÃÃo de PWV-GUA. AnÃlises filogenÃticas das seqÃÃncias dos fragmentos de cDNA correspondentes Ãs capas protÃicas (CP), amplificados a partir dos genomas dos isolados virais de maracujazeiro e de feijoeiro caupi por IP-RT-PCR, agruparam-se com as seqÃÃncias de isolados virais de referidas culturas depositadas no GenBank, apresentando um agrupamento em funÃÃo da especificidade de hospedeiros. Com base nos resultados dos estudos biolÃgicos, sorolÃgicos e, sobretudo moleculares, os isolados virais estudados foram classificados em dois biÃtipos: BiÃtipo CABMV-C (Cowpea) incluindo os isolados obtidos de feijoeiro caupi e biÃtipo CABMV-P (Passion fruit) para incluir os isolados responsÃveis pelo endurecimento dos frutos do maracujazeiro no Brasil.
Santos, Patrícia Resplandes Rocha dos. "Patogenicidade de fungos associados à sementes de andropogon e caracterização morfológica e molecular de Curvularia lunata." Universidade Federal do Tocantins, 2016. http://hdl.handle.net/11612/384.
Повний текст джерелаAndropogon L. is a forage grass, widely distributed in Cerrado areas and with prolonged drought periods. It presents dense characteristics with large clumps and with plumose inflorescence, with high capacity in the seeds dissemination. In turn, seeds are considered the main sources of shelter and transport of pathogens to disease-free areas. In Tocantins, there are no studies records relate the fungi incidence in Andropogon grass seeds as diseases cause in agricultural importance crops. Likewise, there is no research about the transport, transmissibility and pathogenicity of fungi associated with their seeds. The work aim was evaluate the sanitary quality of Andropogon grass seeds, the fungi transmission by seed-seedlings and the fungis pathogenicity to other species plants of agricultural importance, and also perform the Curvularia sp. morphological and molecular characterization, isolated from Andropogon grass seeds. The experiments were conducted at the Phytopathology Research Laboratory, Biologic Control of Disease Laboratory and green house of Tocantins Federal University. In chapter 1, the blotter test method was used to evaluate seed health with and without disinfestation. The fungi incidence was evaluated from seeds individual analysis using stereoscopic and optical microscope. The seeds germination was evaluated after 10 days after test installation, together with the fungi identification associated with non-germinated seeds. For the detected fungus in the sanitary analysis was evaluated the transmission capacity of seed-seedling. The fungi pathogenicity from the Andropogon seeds grass was evaluated by inoculation in the plant itself and was also evaluated the ability of these fungi to infect other plants of economic interest. In chapter 2, the morphological identification was performed from macro and micromorphological observations using as basis the characteristics described in literature regarding the aspect of the colony and conidia of Curvularia sp. Molecular characterization was performed from DNA extraction, amplification and sequencing gene Clg2p region. Transmission was evaluated from seed sowing whitout treatment with fungicides, where at the end of 40 days typical leaf spot symptoms of Curvularia. The pathogenicity was evaluated from the inoculation of conidia suspension on leaves of healthy plants, observing at the end 10 days, if there were symptoms of the pathogen. Were identified and quantified in the seeds of Andropogon L. fungi of the genera Alternaria sp., Bipolaris sp., Curvularia sp., Fusarium sp., Phoma sp., Aspergillus sp., Cladosporium sp., Penicillium sp. and Rhizopus sp. The seed disinfestation reduced the fungi present in the seeds. The fungus Curvularia sp. it was transmitted seed to Andropogon plant. Andropogon L. seeds carried and spread fungi the once inoculated caused infection in the plant itself and other economically important crops, such as rice, cowpea, watermelon, melon, corn, sorghum and grasses marandu, mombaça, piatã and quicuia. Based on morphological and molecular markers, the fungus identified with high incidence associated with Andropogon seeds collected in different agricultural producing regions, is Curvularia lunata. C. lunata is transmitted to plants of Andropogon by seed, being pathogenic to this species of forage grass, causing foliar necrotic spots.
Cisek, Katryna. "Rational Optimization of Small Molecules for Alzheimer’s Disease Premortem Diagnosis." The Ohio State University, 2012. http://rave.ohiolink.edu/etdc/view?acc_num=osu1338325484.
Повний текст джерелаStolf, Beatriz Simonsen. "Identificação de marcadores moleculares para o câncer de tireóide por cDNA microarrays." Universidade de São Paulo, 2003. http://www.teses.usp.br/teses/disponiveis/46/46131/tde-21072008-101124/.
Повний текст джерелаThyroid diseases are very common and are usually benign. The causal relationships among the different types of disease, as well as their molecular aspects, are not well understood. The goiter (hyperplasia), for instance, is described by some as related to papillary carcinoma (a malignant tumor), while others say there is no causal relationship between the two diseases. The most defying question, however, concerns the distinction between adenoma (benign tumor) and follicular carcinoma, which is currently made only after surgery, not allowing distinct treatments for the two kinds of tumor. This work aimed to identify differentially expressed genes among normal thyroid tissue, goiter, adenoma and papillary carcinoma using microarrays. Follicular carcinomas were not included due to the reduced number and size of the samples. Two kinds of array were used: arrays in nylon membranes, with 213 clones isolated from thyroid samples by differential display (DDRT-PCR); and glass slide arrays containing 3800 ORESTES clones.Experiments using the first type of array identified three differentially expressed genes, whose expression was analyzed by RT-PCR in 10 samples of each kind of tissue. Two of these genes were able to differentiate papillary carcinomas from goiters and normal tissues with precisions of 89% for the malignant tumor and 80% for the non-malignant tissues. Glass slide arrays were used to evaluate gene expression profile of approximately 10 samples of each type of thyroid tissue. 160 clones differentially expressed between any two tissues were identified, and their sequences were determined and compared with databases. Among the most interesting genes are Na/K ATPase gene, whose expression is reduced in carcinomas compared to normal tissues and adenomas, the gene corresponding to PDCD4 protein, involved in program cell death, with elevated expression in adenomas and normal tissues than carcinomas and goiters, and the genes of calgizzarin (S100A11) and α1-antitrypsin, both more active in carcinomas than the other tissues. All these genes have already been described as differentially expressed in at least one type of human cancer. This work led to the standardization of glass slide microarray technology in our laboratory, and to the identification of genes that may clarify the alterations involved in the formation of goiter, adenoma and follicular carcinoma. The implementation of mRNA amplification technique in our laboratory allowed the utilization of 10 samples of follicular carcinoma, whose mass was insufficient for microarray hybridizations. These samples will be hybridized along with 10 samples of adenomas, with microarrays containing 4800 known human genes to search for differentially expressed genes, of great diagnostic interest.
Edvinsson, Benjamin. "Molecular diagnosis of infection with Toxoplasma gondii in immunocompromised patients /." Stockholm, 2006. http://diss.kib.ki.se/2006/91-7140-877-0/.
Повний текст джерелаAlhammadi, Mohammed Ali. "Molecular diagnosis and surveillance of community acquired respiratory viral infections." Thesis, University of Manchester, 2008. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.493520.
Повний текст джерелаOwen, Martin Richard. "Molecular approaches to the epidemiology and diagnosis of ovine toxoplasmosis." Thesis, University of Liverpool, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.243214.
Повний текст джерелаHughes, Kelvin J. D. "Molecular methods for the diagnosis of fungal quarantine plant pathogens." Thesis, University of Bristol, 2002. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.272031.
Повний текст джерелаHudspith, Karl Alexander Zhivojin. "Application of genomic technologies for molecular diagnosis of genetic diseases." Thesis, University of Oxford, 2015. https://ora.ox.ac.uk/objects/uuid:b4addaed-e9f9-4762-846a-87eb73f77235.
Повний текст джерелаAraújo, Edivânio Rodrigues de. "Diagnose molecular e estudos epidemiológicos da mancha-bacteriana do tomateiro." reponame:Repositório Institucional da UnB, 2014. http://repositorio.unb.br/handle/10482/16847.
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A mancha-bacteriana está entre os fatores que prejudicam a produção de tomate no Brasil. A doença é causada por um complexo de espécies do gênero Xanthomonas: X. euvesicatoria, X. vesicatoria, X. perforans e X. gardneri. Os objetivos dessa tese são: i) realizar um levantamento da frequência de ocorrência de espécies e raças causadoras da doença em lavouras brasileiras de tomate, tanto destinadas ao consumo in natura quanto ao processamento industrial; ii) estabelecer protocolos de detecção e identificação simultânea das espécies causadoras da doença; iii) compreender o efeito da temperatura e molhamento foliar no monociclo da doença; iv) identificar possíveis agentes causadores da mancha-bacteriana e a comunidade bacteriana presente em mudas assintomáticas cultivadas em viveiro comercial e sementes comerciais de tomateiro. Para alcance desses objetivos, testes de patogenicidade, técnicas moleculares baseadas na PCR (BOX-PCR, iniciadores específicos), além de sequenciamento do gene de avirulência avrXv3 foram utilizados. O efeito da temperatura (15; 20; 25; 30 e 35°C) e molhamento foliar (0; 6; 12; 24 e 48 horas) na severidade da doença foi avaliado para as duas espécies de maior ocorrência: X. perforans e X. gardneri. Da mesma forma, foi verificado o efeito in vitro da temperatura na taxa de multiplicação em meio de cultura dessas espécies. Por fim, por meio de coletas de água superficial no filoplano de mudas assintomáticas e maceração de sementes, buscou-se identificar possíveis agentes patogênicos, tentando identificá-los por isolamento direto, uso de iniciadores específicos e sequenciamento do rRNA de 16S. Com base nos resultados obtidos, verificou-se que X. perforans é a espécie predominante (≈ 92% dos isolados) nos campos brasileiros de produção de tomate, em ambos os segmentos produtivos: consumo in natura e processamento industrial. Entre os isolados de X. perforans, a raça T3 predomina, representando 97,3% desses isolados. Cinco isolado foram identificados como raça T4. Esses isolados apresentaram uma inserção de 858 pb no gene avrXv3, relacionada à família de proteínas YscJ/HrcJ. Por meio dos protocolos estabelecidos foi possível identificar as quatro espécies causadoras da doença com iniciadores específicos tanto utilizando PCR convencional como PCR multiplex. As espécies X. perforans e X. gardneri apresentaram comportamento distinto com relação ao ótimo de temperatura para seu desenvolvimento. Enquanto X. perforans incitou maior porcentagem de área foliar lesionada quando exposta a temperaturas acima de v 25°C, com agressividade crescente até o limite superior testado (35°C), observou-se para X. gardneri uma redução da agressividade nos limites inferior (15°C) e superior de temperatura. Essas espécies só incitaram doença quando o período de molhamento foliar foi igual ou maior que 12 horas. Do mesmo modo, a temperatura teve efeito direto na taxa de crescimento in vitro de X. perforans. Por fim, foi possível identificar um isolado de X. euvesicatoria associado a mudas assintomáticas de tomateiro, além dos gêneros Agrobacterium, Enterobacter, Microbacterium, Sphingobium e Sphingomonas. Com isso, estratégias de manejo da doença devem focalizar X. perforans, já que é a espécie predominante. No mesmo sentido, estratégias de controle denominadas evasivas devem ser adotadas, uma vez que a doença apresenta faixas limítrofes para as variáveis temperatura e molhamento foliar. Este é o primeiro registro da ocorrência da raça T4 no país. Os isolados bacterianos dos demais gêneros encontrados em sementes e mudas devem ser avaliados quanto ao seu potencial como agentes de controle biológico. Levantamentos sistemáticos das espécies/raças causadoras da doença devem ser continuados para o auxílio nas tomadas de decisão. _______________________________________________________________________________________ ABSTRACT
Bacterial spot is among the limiting factors in tomato production in Brazil. The disease is caused by a species complex of the genus Xanthomonas: X. euvesicatoria, X. vesicatoria, X. perforans and X. gardneri. The aims of this thesis were: i) to perform a survey the frequency of occurrence of species and races causing bacterial spot on Brazilian tomato crops; ii) to establish protocols for simultaneous detection and identification of the species causing disease; iii) to understand the effect of temperature and leaf wetness on the monocycle of tomato bacterial spot; iv) to identify possible causal agents of tomato bacterial spot and bacterial community present in asymptomatic seedlings and seeds. For achieving these objectives, pathogenicity tests, molecular techniques based on PCR (BOX-PCR, specific primers) and sequencing of avirulence gene avrXv3 were used. The effect of temperature (15, 20, 25, 30, and 35°C) and leaf wetness (0, 6, 12, 24, and 48 hours) on disease severity was assessed. Likewise, the in vitro effect of temperature on the population growth rate of X. perforans and X. gardneri was checked. Finally, by collecting surface water on the phylloplane of asymptomatic seedlings and soaking commercial seeds, the presence of potential pathogens were surveyed. Direct isolation, using PCR with specific primers and sequencing of 16S rRNA were the tempted methods applied. Based on the results, we found that X. perforans is the predominant species (≈ 92% of isolates) in Brazilian tomato fields, in both production segments: fresh market and processing. Among strains of X. perforans, the T3 race was predominant, representing 97.3% of these isolates. Five isolates were identified as T4 race. These isolates had an insertion of 858 bp in avrXv3 gene related to a protein YscJ/HrcJ family. The protocols enabled the identification of the four species causing the disease with specific primers using both conventional PCR and multiplex PCR. The species X. perforans and X. gardneri showed different behaviors with respect to the optimum temperature for development. Xanthomonas perforans caused more disease at temperatures above 25°C, with increasing aggressiveness up to the upper limit tested (35°C). On the other hand, X. gardneri decreased the disease severity from the optimal temperature for both lower and upper limit of temperature. Both species only incited disease vii when leaf wetness periods were longer than six hours. In the same way, the temperature had a direct effect on the in vitro growth rate of X. perforans. Finally, it was possible to identify an isolate of X. euvesicatoria associated with asymptomatic tomato seedlings in a commercial nursery production, besides the genus Agrobacterium, Enterobacter, Microbacterium, Sphingomonas and Sphingobium. Thus, disease management strategies should focus on the species X. perforans, since it is the predominant species. Similarly, control strategies based on evasion should be applied, since the disease has a borderline range for the temperature and leaf wetness. This is the first report of occurrence of race T4 in Brazil. Bacterial isolates from other genus found in seeds and seedlings should be evaluated to assess their biological control potential. Systematic surveys of species/races causing the disease should be continued to aid in the control decision taking.
Drury, Sarah L. "Molecular, morphological, and kinetic diagnosis of human preimplantation embryo vitality." Thesis, University of Warwick, 2016. http://wrap.warwick.ac.uk/88622/.
Повний текст джерелаINGLESE, NADIA. "Hepatitis E virus from molecular diagnosis to in vitro replication." Doctoral thesis, Università degli Studi di Roma "Tor Vergata", 2009. http://hdl.handle.net/2108/1122.
Повний текст джерелаHepatitis E virus (HEV) is a major cause of acute sporadic and epidemic viral hepatitis in several developing countries where sanitation is suboptimal, but it also occurs sporadically in many developed countries. HEV infects several animal species, mostly asymptomatically. HEV genotype 3 has been shown to have a high prevalence in pigs across Europe and to be closely related genetically to sympatric human HEV strains in developed regions, suggesting both a zoonotic and a possible foodborne transmission. In the first study, the prevalence of swine HEV in pigs from six farms in Northern Italy has been investigated, testing viral RNA in stools by nested RT-PCR. All farms were positive for HEV, with a prevalence ranging between 12.8% to 72.5%. The genome sequences identified were compared with HEV sequences of different origins reported worldwide followed by phylogenetic analysis. In particular, one group of seven Italian strains clustered close to human and swine European HEV strains (91.6-96.2% identity). The lack of a readily available virus propagation system for HEV has greatly hampered investigation of potential transmission routes and detailed analysis of the virus replication cycle in infected cells. In the second study, the rotary cell culture system (RCCS) has been used for in-vitro cultivation of HEV. RCCS is an optimized cell culture technology designed for growing cells in 3D under conditions that promote many features of in-vivo tissues. Cells used in these studies were: human intestinal epithelium interferon knocked-out (Int-407 IFN KO) engineered to constitutively block the cellular antiviral response, porcine kidney cells (PK-15 IFN KO), human hepatoma cell line (PLC/PRF/5) and Hep G2/C3a. These cell lines were inoculated with HEV derived from PCR-positive swine faecal suspensions, pig liver tissue and liver culture samples from experimentally infected pigs. Real time PCR was performed to detect and quantify HEV RNA in the cultures throughout the observation period and also to evaluate, together with RT-PCR, IPX, CM, SEM and TEM, differences in the expression of cell differentiation/infection markers (CK-8, CK-18, CK-19, IL-6, CytP450 and Albumin) between the cultures in 3D configuration compared to conventional 2D cultures of the same cells. The results suggest that the 3D cell aggregates show many of the specialized features of in vivo tissues offering a novel approach for studying viral replication and pathobiology compared to conventional 2D analysis of the same cells.