Дисертації з теми "Nucleic-acid Amplification and Quantification"
Оформте джерело за APA, MLA, Chicago, Harvard та іншими стилями
Ознайомтеся з топ-50 дисертацій для дослідження на тему "Nucleic-acid Amplification and Quantification".
Біля кожної праці в переліку літератури доступна кнопка «Додати до бібліографії». Скористайтеся нею – і ми автоматично оформимо бібліографічне посилання на обрану працю в потрібному вам стилі цитування: APA, MLA, «Гарвард», «Чикаго», «Ванкувер» тощо.
Також ви можете завантажити повний текст наукової публікації у форматі «.pdf» та прочитати онлайн анотацію до роботи, якщо відповідні параметри наявні в метаданих.
Переглядайте дисертації для різних дисциплін та оформлюйте правильно вашу бібліографію.
Hernández-Neuta, Iván. "Nucleic acid analysis tools : Novel technologies and biomedical applications." Doctoral thesis, Stockholms universitet, Institutionen för biokemi och biofysik, 2017. http://urn.kb.se/resolve?urn=urn:nbn:se:su:diva-146334.
Повний текст джерелаAt the time of the doctoral defense, the following paper was unpublished and had a status as follows: Paper 1: Manuscript.
Lee, Dong-Hun. "Nucleic acid amplification testing for screening of individual blood units." [S.l. : Amsterdam : s.n.] ; Universiteit van Amsterdam [Host], 2007. http://dare.uva.nl/document/48208.
Повний текст джерелаDaher, Rana. "Recombinase polymerase amplification technology : Assessment for nucleic acid-based acid-based point-of-care diagnostics." Doctoral thesis, Université Laval, 2015. http://hdl.handle.net/20.500.11794/26269.
Повний текст джерелаThis dissertation consists of an exhaustive study on an emerging technology for isothermal amplification of nucleic acids called recombinase polymerase amplification (RPA). The introduction of this thesis is a detailed description of the RPA. This review documents and discusses the various applications of this technology by pointing to the current knowledge about RPA for diagnostic applications. Despite the complex composition of RPA (6 to 7 proteins in the same reaction mixture), the latter was shown to be rapid (generating results in < 20 min), specific and sensitive (detecting few target genome copies), and applied widely in different fields. Based on these advantages, we assume that RPA has a flexibility allowing it to be used for the rapid diagnosis of infectious diseases thus reducing time-to-result to less than an hour. Consequently, it will be possible to integrate RPA in microfluidic platforms providing a lab-on chip system. The first part of this doctoral project generated additional guidelines for RPA primers/probes design to develop specific RPA diagnostic assays. Second, we developed an RPA diagnostic test for the detection of group B streptococci, responsible for sepsis and meningitis in newborns. This assay was the first to evaluate RPA with human clinical samples. This diagnostic test was compared to a reference method, the polymerase chain reaction (PCR). This demonstration with clinical samples served to carry out the final objective of this project that was to automate RPA in a miniaturized microfluidic centripetal system. Collaboration with engineers and experts in materials has generated the microfluidic device called "blade" and the instrument involved in the operation of various mechanistic tasks. These preliminary results suggested that it will be important to provide an automated system applicable at bedside. Consequently, it will be possible to perform a complete analysis of infectious agents in less than an hour without the need for complex procedures for the preparation and transport of clinical specimens or the assistance of qualified personnel.
Matinyenya, Brian. "Novel and newer nucleic acid amplification tests for the diagnosis of TB." Master's thesis, University of Cape Town, 2016. http://hdl.handle.net/11427/20680.
Повний текст джерелаSyed, Shahida Nina. "Electrochemical control of reversible DNA hybridisation : for future use in nucleic acid amplification." Thesis, University of Edinburgh, 2014. http://hdl.handle.net/1842/9617.
Повний текст джерелаThomas, Alistair Owen. "Detection of bacterial gene expression by a novel isothermic nucleic acid amplification technology." Thesis, University of Bath, 2004. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.410924.
Повний текст джерелаXiao, Linlin. "Detection of Viable Foodborne Pathogens and Spoilage Microorganisms by Nucleic Acid Amplification Based Platforms." The Ohio State University, 2011. http://rave.ohiolink.edu/etdc/view?acc_num=osu1308284180.
Повний текст джерелаCORAL, LUCIA. "HIGH-RESOLUTION NUCLEIC ACID ANALYSIS WITH A DNA NANOTECHNOLOGY APPROACH." Doctoral thesis, Università degli Studi di Trieste, 2017. http://hdl.handle.net/11368/2908115.
Повний текст джерелаChoi, Kwan-yue. "A molecular epidemiology study on conjunctivitis using conventional nucleic acid amplification technologies and resequencing microarray." Click to view the E-thesis via HKUTO, 2009. http://sunzi.lib.hku.hk/hkuto/record/B44248465.
Повний текст джерелаChoi, Kwan-yue, and 蔡君如. "A molecular epidemiology study on conjunctivitis using conventional nucleic acid amplification technologies and resequencing microarray." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2009. http://hub.hku.hk/bib/B44248465.
Повний текст джерелаBowers, Katherine. "Development and clinical performance of nucleic acid amplification techniques for the diagnosis of Strongyloides stercoralis." Thesis, University of Westminster, 2017. https://westminsterresearch.westminster.ac.uk/item/q56v6/development-and-clinical-performance-of-nucleic-acid-amplification-techniques-for-the-diagnosis-of-strongyloides-stercoralis.
Повний текст джерелаHenriksson, Sara. "Application of Padlock Probe Based Nucleic Acid Analysis In Situ." Doctoral thesis, Uppsala universitet, Institutionen för genetik och patologi, 2010. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-128446.
Повний текст джерелаLehnus, Massimiliano. "Bio-BCA (Bio-Barcode Cascade Amplification) : development of a photosensitive, DNA-based exponential amplification platform technology for the detection of nucleic acid biomarkers." Thesis, University of Cambridge, 2018. https://www.repository.cam.ac.uk/handle/1810/277915.
Повний текст джерелаDann, Louise Claire. "Nucleic acid sequence-based amplification : relative performance and applications in HIV-1 disease monitoring and patient management." Thesis, University College London (University of London), 2002. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.272347.
Повний текст джерелаGrant, Paul Robert. "Development and application of nucleic acid amplification technology (NAT) for the detection of viruses in donated blood." Thesis, University College London (University of London), 2003. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.408734.
Повний текст джерелаGarneret, Pierre. "Microfluidique papier pour le diagnostic de terrain : préparation d'échantillon et multiplexage." Thesis, Paris Sciences et Lettres (ComUE), 2019. https://pastel.archives-ouvertes.fr/tel-03174261.
Повний текст джерелаSince the Ebola outbreak of 2014, the MMN Laboratory and the Pasteur Institute are working to conduct molecular biology tests on paper microfluidics for the diagnosis of infectious diseases. Tests made in Guinée in 2015, by the two teams, have shown the pertinence of the technology. Today, both teams are working on developing a multiplexed devices (simultaneous detection of multiple biological targets). The recent Zika Virus (ZIKV) outbreak transmitted by Aedes mosquitoes in geographic area where other arboviruses such as Dengue (DENV) and Chikungunya (CHIKV) were already spreading focused the development work on a Zika/Dengue/Chikungunya multiplexed test. The first goal is to get a point of care device, cheap and easy to use allowing a better management of the patients. The development of such a device would then be used by Pasteur Institut network to monitor more precisely the epidemiology of those diseases displaying the same the same symptomatology, and study the physiology of their co-infection
Masetty, Manaswini. "A Smartphone Enabled Molecular Diagnostic Toolkit to Detect Pathogens via Isothermal Nucleic Acid Amplification on Pre-Dried Disposable Paper Strips." University of Cincinnati / OhioLINK, 2021. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1627664394713446.
Повний текст джерелаHofmann, Jakob [Verfasser]. "Evaluation der klinischen Praktikabilität und prognostischen Aussagefähigkeit der intraoperativen Sentinellymphknotendiagnostik mittels One-step Nucleic Acid Amplification (OSNA) bei invasivem Mammakarzinom / Jakob Hofmann." Kiel : Universitätsbibliothek Kiel, 2014. http://d-nb.info/1053326211/34.
Повний текст джерелаBernander, Sverker. "Detection and epidemiologic subtyping of Legionella pneumophila using DNA-based molecular methods /." Stockholm, 2003. http://diss.kib.ki.se/2003/91-7349-745-2.
Повний текст джерелаNicolini, Ariana Marie, and Ariana Marie Nicolini. "Single-Step, Optical Biosensors for the Rapid and Sensitive Detection of Bacterial and Viral Pathogens." Diss., The University of Arizona, 2016. http://hdl.handle.net/10150/623158.
Повний текст джерелаFaltin, Bernd [Verfasser], and Roland [Akademischer Betreuer] Zengerle. "Mediator Probe PCR: a novel assay principle for universal real-time detection of nucleic acid amplification = Mediator Probe PCR: ein neuartiger Ansatz zur universellen Echtzeit-Detektion von Nukleinsäuren." Freiburg : Universität, 2013. http://d-nb.info/1123477000/34.
Повний текст джерелаHadersdorfer, Johannes [Verfasser], Dieter Richard [Akademischer Betreuer] Treutter, and Thilo [Akademischer Betreuer] Fischer. "Development of an isothermal nucleic acid amplification protocol for high-throughput monitoring of Plum pox virus infection in stone fruit production / Johannes Hadersdorfer. Gutachter: Dieter Richard Treutter ; Thilo Fischer. Betreuer: Dieter Richard Treutter." München : Universitätsbibliothek der TU München, 2013. http://d-nb.info/1032313498/34.
Повний текст джерелаGomez, Deborah Beltrami. "Prevalência de Chlamydia trachomatis em mulheres inférteis e gestantes assintomáticas." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2016. http://hdl.handle.net/10183/143385.
Повний текст джерелаBackground: Chlamydia trachomatis (CT) is the most prevalent sexually transmitted bacterial infection and affects mainly young, sexually active, women. Untreated infection may lead to reproductive complications due to tubal damage. Infections during pregnancy may cause preterm labor, low birth weight, perinatal death and neonatal conjunctivitis and pneumonia. There is little data on CT infection in Brazil. The aim of this study was to determine CT prevalence on infertile and pregnant women. Methods: A cross-sectional study included 77 infertile and 60 asymptomatic pregnant women. First void urine was tested to CT using PCR and blood samples were collected for CT IgG antibodies testing using Indirect Immunofluorescence. A questionnaire about medical, gynecological and sexual history was applied to all participants. Results: We found statistically similar prevalence of PCR and IgG antibodies between groups. This study observed a 61% prevalence of CT IgG antibodies in infertile women and 56,7% in pregnant women. PCR was positive in only one (1,3%) infertile woman and in none of the pregnant. Conclusion: A high prevalence of C. trachomatis IgG antibody in Brazilian pregnant and infertile women, but a low prevalence of positive PCR on urine samples were demonstrated. CT antibodies were associated with sexual behavior and smoking.
Pugnière, Pascal. "Contribution à l'amélioration de la quantification des acides nucléiques par qPCR et RT-qPCR." Phd thesis, Université de Grenoble, 2012. http://tel.archives-ouvertes.fr/tel-00870512.
Повний текст джерелаMaree, Hans Jacob. "Identification and characterisation of grapevine leafroll-associated virus 3 genomic and subgenomic RNAs." Stellenbosch : University of Stellenbosch, 2010. http://hdl.handle.net/10019.1/5417.
Повний текст джерелаIncludes bibliography.
Title page: Dept. of Genetics, Faculty of Science
ENGLISH ABSTRACT: Grapevine leafroll-associated virus 3 (GLRaV-3) is the type strain for the genus Ampelovirus, family Closteroviridae. There has been only one report that claimed the complete nucleotide sequence of GLRaV-3 (isolate NY-1, AF037268). Here we report the complete sequence of the South African GLRaV-3, isolate GP18 (EU259806) and show a significantly extended 5’ end. We used RLM-RACE to determine the 5’ end of GP18 and found the 5’ UTR to be 737 nt compared to 158 nt in the NY-1 sequence. This extended UTR was found in all other South African isolates of GLRaV-3 that were tested. In two collaborative studies the existence of the extended 5’ UTR was confirmed and further investigated. In the first study (Coetzee et al., 2010), metagenomic data generated by next generation sequencing (Illumina Genome Analyzer II) was analysed for GLRaV-3 specific sequences. Sequences similar to the GP18 isolate confirmed the sequence of the extended 5’ UTR. In the second study (Jooste et al., 2010), three genetic variants were identified and their respective 5’ UTRs studied. Great diversity was observed between the 5’ UTRs of the different genetic variants, however within a variant the 5’ UTR was found to be highly conserved. Grapevine leafroll-associated virus 3 is a positive sense, single stranded RNA virus that has been shown, like other closteroviruses, to produce subgenomic (sg) RNAs during replication. These sgRNAs are deployed for the expression of the ORFs on the 3’ half of the genome. In this study a dsRNA blot confirmed the presence of three, 3’ coterminal sgRNAs species [sgRNA(ORF3/4), sgRNA(ORF5) and sgRNA(ORF6)] in GLRaV-3-infected plant material when using a probe directed at the coat protein gene. The specific 5’ terminal nucleotides for these sgRNAs as well as four additional sgRNAs [sgRNA(ORF7), sgRNA(ORF8), sgRNA(ORF9) and sgRNA(ORF10-12)] were determined by RLM-RACE for GLRaV-3 isolate GP18. The construction of a GLRaV-3 mini-replicon, analogous to RNA1 of Lettuce infectious yellows virus, for the evaluation of putative sg-promoters is also described.
AFRIKAANSE OPSOMMING: Grapevine leafroll-associated virus 3 (GLRaV-3) is ‘n lid van die Closteroviridae familie en die hooflid vir die genus Ampelovirus. Tot dusver was daar net een studie wat die volledige nukleïensuurvolgorde van GLRaV-3 gerapporteer het (isolaat NY-1, AF037268). In hierdie studie rapporteer ons die volledige volgorde van ‘n Suid-Afrikaanse GLRaV-3, isolaat nl. GP18 (EU259806) wat noemenswaardig langer is aan die 5’ kant. RLM-RACE is gebruik om die 5’ eindpunt van GP18 te bepaal en daar is gevind dat die 5’ ongetransleerde streek (UTR) 737 nt lank is in vergelyking met die 158 nt van die NY-1 volgorde. Die verlengde 5’ UTR is gevind in alle Suid-Afrikaanse monsters wat getoets is. Die verlengde 5’ UTR is bevestig en verder bestudeer tydens twee samewerkingsprojekte. In die eerste studie (Coetzee et al., 2010), is metagenomiese data gegenereer deur volgende-generasie volgordebepaling (Illumina Genome Analyzer II) en geanaliseer vir GLRaV-3 spesifieke volgordes. Volgordes soortgelyk aan die GP18 isolaat het die verlengde 5’ UTR volgorde bevestig. In die tweede studie (Jooste et al., 2010), is drie genetiese variante van GLRaV-3 geidentifiseer en hulle onderskeie 5’ UTR volgordes bepaal en bestudeer. Daar is groot diversiteit tussen die 5’ UTRs van die verskillende genetiese variante gevind, maar tussen isolate van dieselfde variant is die volgordes gekonserveerd. Grapevine leafroll-associated virus 3 is ‘n positiewe-sin, enkelstring RNA virus wat al voorheen bewys is om, soos ander closterovirusse, subgenomiese (sg) RNAs te produseer tydens replisering. Hierdie sgRNAs word ingespan vir die uitdrukking van die ORFs op die 3’ helfte van die virusgenoom. In hierdie studie is ‘n dsRNA klad gebruik om die voorkoms van 3’ ko-terminale sgRNAs [sgRNA(ORF3/4), sgRNA(ORF5) and sgRNA(ORF6)] te bevestig in GLRaV-3 geinfekteerde plantmateriaal deur gebruik te maak van ‘n peiler teen die kapsiedproteïengeen. Die spesifieke 5’ terminale nukleotiedes vir hierdie sgRNAs sowel as vier additionele sgRNAs [sgRNA(ORF7), sgRNA(ORF8), sgRNA(ORF9) and sgRNA(ORF10-12)] is bepaal deur gebruik te maak van RLM-RACE op die GLRaV-3 isolaat GP18. Die konstruksie van ‘n GLRaV-3 mini-repliserings konstruk, analoog aan die RNA1 van Lettuce infectious yellows virus, vir die evaluasie van moontlike sg-promotors word ook beskryf.
Gualberto, Felipe Augusto Souza. "Valor disgnóstico da nested PCR em tempo real em pacientes com meningite tuberculosa." Universidade de São Paulo, 2014. http://www.teses.usp.br/teses/disponiveis/5/5134/tde-26082014-093325/.
Повний текст джерелаBackground: Tuberculous meningitis (TBM) is the most serious and lethal presentation of tuberculosis. Timely diagnosis and appropriated treatment are the main factors associated with good outcome. Methods used in the daily medical practice - clinical, radiological and cerebrospinal fluid (CSF) findings - have low accuracy. Search for Mycobacterium tuberculosis DNA in the CSF by polymerase chain reaction (PCR) using the nested methodology is promising, especially when combined with the practical approach of the real time DNA amplification. Objective: To evaluate the diagnostic value of a nested real-time PCR (nRT-PCR) in the investigation of patients with TBM. Methods: A two-phase observational study was carried out: prospective and retrospective. In the prospective phase, patients with suspected TBM hospitalized at \"Instituto de Infectologia Emílio Ribas\" (IIER) were included. Clinical, laboratory and radiological data were collected, as well as CSF samples of all patients. According to international standard criteria, patients were categorized as \"TBM Definite\", \"TBM Probable\", \"TBM Possible\" and \"Not TBM\". The nRT-PCR, using the mpt64 gene, was performed on all CSF sample in the Laboratory of Bacterial Meningitis, Adolfo Lutz Institute. Sensitivity, specificity and confidence intervals (95% CI) of the nRT-PCR were calculated based on the gold standard (culture positive for M. tuberculosis or AFB isolation on the central nervous system) and on patients with other established diagnoses (\"Not TBM\"). The proportion of patients with a positive nRT-PCR in each clinical category was also calculated. In the retrospective phase, medical chart review was performed in those patients who had the nRT-PCR requested in IIER and in the \"Centro de Referência e Treinamento em DST/AIDS\". The same diagnostic categorization and calculations of sensitivity and specificity were adopted. Results: 102 patients were included in the prospective phase, 92 of them HIV-infected. Nine of them had the gold standard positive and were classified as \"TBM Definite\" and 81 of them had other diagnoses established (\"Not TBM\"). The sensitivity and specificity of the nRT-PCR were 100% (95%CI: 70-100 and 95-100, respectively). The nRT-PCR positivity in category \"TBM Probable\" was 50% (4/8 patients) and 25% in \"TBM Possible\" (1/4). In retrospective phase, the nRT-PCR had a sensitivity of 83% (5/6) and specificity of 100% (0/45), among the 56 included patients (48 of them HIV infected). Positivity in \"TBM Probable\" category was 60% (3/5) and no patients were classified as \"TBM Possible\". Conclusion: The nRT-PCR showed good sensitivity and excellent specificity, showing its diagnostic value in the timely identification of TBM
Priyanka, V. "Droplet Isothermal Amplification For Nucleic Acid Quantification." Thesis, 2022. https://etd.iisc.ac.in/handle/2005/5643.
Повний текст джерелаJue, Erik Bradley. "Improved Tools for Point-of-Care Nucleic Acid Amplification Testing." Thesis, 2020. https://thesis.library.caltech.edu/13735/1/200529_erik_jue_2020_thesis_final.pdf.
Повний текст джерелаHsieh, Tsung-Min, and 謝宗閔. "Micro Thermocyclers for Nucleic Acid Amplification with High Thermal Uniformity." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/30145900653046174870.
Повний текст джерела國立成功大學
電機工程學系碩博士班
96
The trend of miniaturization of biomedical instruments began in the last century, and is crucial for furthering progress in medical technology. In this study, an integrated chip–PCR/RT-PCR system is implemented successfully, meeting the requirements for use in miniature thermocyclers – small in size, low power consumption, high heating and cooling rates, and even portability, with resistance temperature detectors and thin-film micro-heaters. Furthermore, due to the modifications made to the micro thermocycler, the thermal uniformity of a specific region has been greatly enhanced, improving the performance of the micro device. The initial approach of this study was to develop a block-type thin-film micro-heater and resistance temperature detectors on a glass substrate to form a micro thermocycler. Based on the techniques of feedback control and pulse width modulation, PCR thermal cycling was achieved, and the system was tested using Salmonella, with successful results. Two new approaches were then implemented and applied to increase DNA amplification efficiency and reduce non-specific PCR products. In order to achieve these aims, first, a novel kind of micro thin-film heater, an array-type micro-heater, was designed and developed. The main advantages of array-type micro-heaters are their lower resistance as compared with block-type heaters and their provision of greater thermal uniformity with a uniform heating density. Active compensating units were also added and applied in order to reduce the edge-effect, i.e., thermal loss from the edges. From these array-type micro-heaters and active compensating units, a miniature thermocycler with enhanced thermal uniformity was developed and the success of its performance verified with S. pneumoniae. The second approach was the design of a novel self-compensating array-type micro-heater. This kind of micro thin-film heater not only increases the thermal uniformity in the specific heating area, but also eliminates additional control themes on thermal compensating. Based on self-compensating array-type micro-heaters, this device is of a fully two-dimensional thermal compensating design used in thin-film heaters. Experimental results from infrared images showed that the percentage of the uniformity area with a thermal variation of less than 1�aC was 90.3%, 99.9% and 96.8% at PCR thermocycling temperatures of 94�aC, 55�aC and 72�aC, respectively, which represents a significant improvement over conventional block-type or serpentine-shaped micro-heaters. The performance of the chip–PCR system based on self-compensating array-type micro-heaters was assessed by amplifying a detection gene (171bp) associated with the dengue virus serotype 2, the results of which were successful, the amplification efficiency having been found to be statistically greater than that of other micro-heaters without a self-compensation function. The micro-heater proposed in this study was based on the aim of finding an efficient approach to greatly improve the thermal uniformity of a specific micro area requiring precise thermal conditions, and a micro thermocycler for nucleic acid amplification with high thermal uniformity was developed. This system not only satisfies the requirements of micro biomedical devices, but could be also useful in the design of a micro-total-analysis-system and a lab-on-a-chip, which require high thermal uniformity in order to improve the performance of the miniature devices.
Drake, Philip, Y.-C. Chen, I. Lehmann, and P.-S. Jiang. "Nanoparticle labels for pathogen detection through nucleic acid amplification tests." 2014. http://hdl.handle.net/10454/10333.
Повний текст джерелаMagnetic nanoparticles and surface-enhanced Raman scattering (SERS) active nanoparticles were coated with short chain DNA tags. These were then used to identify a target bacterial DNA sequence. The tags function as primers in a standard PCR with the reverse primers and forward primers on the SERS nanoparticles and magnetic nanoparticles, respectively. During the PCR cycles, a composite nanostructure is formed that is both magnetically responsive and SERS active. After magnetic trapping, the intensity of the SERS signal can be related back to the concentration of the target DNA. A test assay was performed that showed a detection limit (based on the signal to noise ratio) of less than 3 zeptomole (41 pg/L). For comparison, a PCR assay based on the standard SYBR Green method was performed. This used the same primers and target DNA and had a detection limit of 10 attomoles (138 ng/L), 3,000 times less sensitive. The work documents the proof of principle study and shows for the first time the use of SERS-NP labels in the quantification of nucleic acid amplification tests and PCR.
Ping-Hua, Teng, and 鄧秉華. "Application of isothermal nucleic acid amplification on shrimp viral disease diagnosis." Thesis, 2009. http://ndltd.ncl.edu.tw/handle/39938024179183265321.
Повний текст джерела東海大學
畜產與生物科技學系
97
Shrimp viral diseases are important issues in aquaculture industries. Until now, there have been not efficient vaccines or therapeutic strategies against these viral diseases. To face the threats, farmers tried to decrease cultivation risks with more efficient methods. However, they had no tools to early detect and prevent viruses into cultivation environment. For the purpose of pathogen detection before disease outbreak, a diagnostic platform with efficiency and accuracy plays an important role for health management of shrimp culture. Traditional strategies, such as histochemistry or immune-related techniques, only could be used as disease determination because of their insufficient sensitivities. To pathogen prevention, however, applications of molecular diagnostic technologies are feasible strategies to conquer the defects of sensitivities and complexities of histochemistry and immuo-related techniques. Polymerase chain reaction (PCR), for example, is a powerful diagnostic tool that can detect even 10 molecules in the reaction. PCR related techniques had been used to establish the culture system of specific pathogen free (SPF) and daily diagnosis for large-scale or integrated farms. However, the needs of equipments and technicians for a PCR laboratory operating are too expensive for medium or small-sized farms. Although some diagnostic centers may provide services of PCR diagnosis, the time to get results is always too long. With so many difficults to get efficient diagnostic tools, the pathogenic threats are accomplished with these smaller farms. In the study, some isothermal amplification techniques will be applied to develop a simple, economic and accurate molecular diagnostic platform to match farmers’ needs. Isothermal amplification techniques, including ramification amplification (RAM), nucleic acid sequence based amplification (NASBA) and loop-mediated isothermal amplification (LAMP), will be evaluated the feasibilities. In addition, real-time detection of these products amplified by isothermal amplifications is also tested to avoid post-amplification procedures and simplify data interpretation. The results showed that the sensitivity of RAM in detecting IHHNV was 100 copies per reaction, and the sensitivity of NASBA and reverse transcription LAMP in detecting TSV was 50 and 100 copies per reaction, respectively. Three kinds of isothermal amplification methods could be achieved high sensitivities comparable to that of PCR at 3 hours. The specificities and simplicities of these methods are also verified under different conditions to match the purpose of on-site use. For the purposes of real-time detection and quantification, LAMP coupled with fluorescence resonance energy transfer (FRET) techniques is studied and the results are available for follow-up developments. For further utilities, a novel real-time isothermal amplification detection machine should be developed to fit the purpose of diagnosis with simple, economic and accurate.
Panwar, Jatin. "Droplet Microfluidics for Nucleic Acid Quantification and Single Cell Analysis." Thesis, 2019. https://etd.iisc.ac.in/handle/2005/5018.
Повний текст джерелаLiao, Chia-Sheng, and 廖家陞. "Automatic Nucleic Acid Amplification Microsystems for Rapid Pathogen Diagnosis of Infectious Disease." Thesis, 2005. http://ndltd.ncl.edu.tw/handle/69312723014061329434.
Повний текст джерела國立成功大學
微機電系統工程研究所
93
Rapid advances in biochemistry and bio-genetics lead to molecular biology, which is revolutionizing scientific thought. A micro-electro-mechanical-system (MEMS) allows human manipulation of micrometer-scale molecules. Advancement in biotechnology will hopefully improve the quality of human life. This investigation presents an automatic rapid diagnosis microsystem based on nucleic acid amplification. The miniature system is fabricated using MEMS techniques, and consists of a micro temperature control module and a microfluidic control module. The heating and temperature sensing elements of micro temperature control module are both fabricated using platinum, and are located within the reaction chambers to generate rapid and uniform thermal cycling. The microfluidic control module, including the reservoir, micropumps, microvalves and microchannel, enables automatic testing with minimal human intervention. This study proposes a microsystem to detect several genes associated with the DNA-based upper respiratory tract infection microorganisms, their corresponding antibiotic-resistant gene and RNA-based virus. An efficiency multiplex amplification of many targets of interests in one reaction is feasible using the proposed micro device. Additionally, multiple PCR, a novel approach of processing of various samples with different thermal cycle conditions in parallel, is developed for fast diagnosis. Multiple PCR is much faster than the traditional bench-top PCR machine system. The proposed microsystem also realizes a two-step reverse transcription polymerase chain reaction. The complete detection process, including cell lysis, sample/reagents transportation, cDNA synthesis and amplification, can be automatically performed in a user-friendly manner. The experimental data demonstrate that the high heating/cooling rates of the microsystem (20℃/sec and 10℃/sec, respectively) permit successful amplification of DNA microorganisms within 15 minutes and of RNA virus within one hour. The portable microsystem is packaged and operated by a 9-volt battery. The developed microsystem provides a crucial tool for many applications such as genetic analysis, molecular biology and infectious disease detection.
Yang, Litao. "Coupling aptamer biosensors to signal amplification." Thesis, 2007. http://hdl.handle.net/2152/3098.
Повний текст джерелаYang, Litao 1976. "Coupling aptamer biosensors to signal amplification." 2007. http://hdl.handle.net/2152/13284.
Повний текст джерелаChou, Wen-Pin, and 周文彬. "Development of a Novel Nucleic Acid Amplification System-Capillary Convective Polymerase Chain Reaction." Thesis, 2011. http://ndltd.ncl.edu.tw/handle/19692785520165047927.
Повний текст джерела國立臺灣大學
機械工程學研究所
99
A typical polymerase chain reaction (PCR) is a process containing three temperature steps for denaturation, annealing and elongation. In a traditional thermal cycler, metal block is used to heat or cool the reaction tubes, and more than two hours are needed for 45 cycles of the repeat heating and cooling steps. However, expensive thermal controller and long time-consuming made thermal cyclers are not suitable in self-test for personal use. In this study we describe a new method for DNA amplification using a principle of convection. In this platform, only a dry bath is required for heating the bottom of the reaction tube. Then the tube is cooled by the surrounding air by which the reagent in the tube forming convection repeatly. Thus, reaction mixture in the tube should undergo the three steps of PCR in the convection. This kind of convection in the tube is just like the phenomenon during water is boiled and it makes the reagents in the tube should fluid spontaneously to achieve different temperatures. This technique can shorten the amplification time prominently and solve the traditional problem for long time-consuming process during heating and cooling. By this way, no expensive equipment is needed and it makes this new method feasible to be used in almost all the laboratories. The key point of successful working of this platform is how to make the fluid reagent mixture of PCR reaction in the tube to form a stable and efficient convection. Here we focus on two important parameters. One is the ratio between height and diameter of the reaction tube, which determines whether the temperature gradient from the bottom to top can fulfill the three-step temperatures needed in the PCR reaction. The other is the criterions of the reagents designed in convective PCR. Because reagents in convective PCR reaction undergo a continued temperature gradient which is different from traditional PCR, it requires particular criterions for reagents in order to improve the level of yield and the rate of success. In this study, we also provide the physical parameters and design criterions of particular reagents for successful convective PCR. Using this newly established method, DNA amplification could be completed within 25 minutes and the sensitivity was measured to reach 30 copies/per tube. Besides, specific reagents were also designed for various virues, including new influenza virus H1N1, white spot syndrome virus (WSSV), and polyomavirus (BKV). Moreover, we also explore whether primers conjugated with LNA should avoid side products in the amplification. We will focus on developing a cheaper and more convenient technique for examination using our established platform on site, where it could be operated by persons without well-training. We also believe this study will provide a new direction and will be helpful and useful in the future in controlling infectious disease for human, animal and plants.
Loan, Thomas. "Cell Lysate as a Platform Technology for Biocatalytic Synthesis and Nucleic Acid Amplification." Phd thesis, 2020. http://hdl.handle.net/1885/204831.
Повний текст джерелаDAI, NING, and 戴寧. "Development of Thermostatic Nucleic Acid Amplification Technology for Detecting Acute Hepatopancreatic Necrosis Disease (AHPND)." Thesis, 2016. http://ndltd.ncl.edu.tw/handle/06651700425244830036.
Повний текст джерела國立高雄海洋科技大學
海洋生物技術研究所
104
The outbreak of acute hepatopancreatic necrosis disease (AHPND) from 2009 has caused mass losses in shrimp farms worldwide. The strain of Vibrio parahaemolyticus which obtain a plasmid and secret toxins has been determined as the infectious agent. PCR methods were published and used to perform diagnosis in laboratory. This study developed methods of Loop-mediated isothermal amplification (LAMP) and Recombinase polymerase amplification (RPA) to detect AHPND and aimed to carry out the diagnosis at the farm side. Compared to PCR, the nucleic acid amplification of LAMP and RPA is performed at a constant temperature which shortens the reaction time and excludes complex equipments. The results are directly color presented by fluorescent dye or lateral flow dipstick which substitute for the gel electrophoresis. The LAMP system for detection of AHPND have six primers targeting toxin A gene (333 bp) in the plasmid under 65 ℃condition. LAMP results were observed through the stain of fluorescent dye and showed the color change emitted by ultraviolet light. RPA have designed two methods for amplification of the target sequence, one is using two labeled primers, the other is added the probe to increase the specificity. After reaction at 37 ℃, both methods followed with lateral flow dipstick which antibody labeled recognize RPA amplicons. Combined with the DNA easy extraction method for template preparation, the time needed to complete the diagnosis is only 1-2 hours. The sensitivity and specificity of LAMP and RPA are better or comparable to PCR in this study. We also examined several white shrimps (Litopenaeus vannamei) from Kaohsiung and found positive signals. It revealed the distribution and consideration of AHPND.
Lien, Kang-Yi, and 連剛逸. "Miniature Microfluidic System Integrated with a Sample Pretreatment Device for Rapid Nucleic Acid Amplification." Thesis, 2007. http://ndltd.ncl.edu.tw/handle/84844620753199418940.
Повний текст джерела國立成功大學
奈米科技暨微系統工程研究所
95
Recently, purification and enrichment of bio-samples is crucial for the analysis of biosamples with an ultra-low concentration that the performance of the detection system can be efficiently increased. Apart from that, clinical samples usually contain biological medium that would normally inhibit the subsequent ribonucleic acid (RNA)/deoxyribonucleic acid (DNA) amplification process. Moreover, there still need several tedious purification and washing steps and labor-intensive processes to complete the sample preparation. In addition, a number of large-scale equipments are usually needed in the complex pretreatment procedures and the bio-samples may also be wasted and cause some contaminations during manual operations. To improve this, the concept of MEMS (micro-electro-mechanical-system)-based miniaturization is applied in the magnet beard-based sample purification and enrichment process. It is generally believed that the miniaturized bio-devices typically bring the advantages of shorter diagnosis times, less sample and reagent consumption, improved resolution, higher sensitivity, and lower cost. By the incorporation of the microfluidic system, the miniature bio-devices provide the powerful techniques to transport or mix the fluids in the system and make the biological diagnosis a quick and automatic process. Nevertheless, the integrated systems still need other detection chambers to perform biological diagnosis. And the on-chip microfluidic modules still need several sets of unidirectional microfluidic structures to complete the whole process. As a result, several electromagnetic valves (EMVs) are usually required to deliver the fluid in the microchannels and the manual operations are normally required since the lack of function of sample pretreatment in the micro system. Consequently, there is a need for a sample pretreatment process by utilizing a microfluidic system to automate the nucleic acid amplification process with less human intervention and also to improve sensitivity and selectivity. The study therefore proposes three new miniature reverse transcription polymerase chain reaction (RT-PCR) systems that integrate with bio-sample pretreatment devices using superparamagnetic beads into a single chip platform. In the first system, three modules were integrated including a microfluidic nodule consists of three sets of novel pneumatic micropumps, a bead collection module with Au wires and a micro temperature control module for the polymerase chain reaction (PCR) process. For the second system proposed, the microfluidic chip integrated two functional devices for bio-samples purification and enrichment including pumping, mixing and separation by utilizing a rotary microfluidic module and a bead-collection module consists of 2-dimension/3-dimension (2-D/3-D) microcoils. The original rotary microfluidic module can provide a rapid flow pumping rate and a high mixing efficiency and can pretreat the bio-samples in a short period of time. The third microsystem uses the novel microfluidic system including a two-way serpentine-shape (s-shape) micropump requiring only one EMV to transport and to mix the bio-samples in the microchannels. In addition, new bio-separators are developed either to perform the separation of magnetic beads or to control the temperature field for the subsequent RT-PCR process. The rapid heating/cooling rate of the micro-heating chambers can significantly shorten the pre-treatment and diagnosis processes. As a whole, the developed system may provide a powerful platform integrating the functions of sample pretreatment and fast disease diagnosis.
Chen, Wei-Jang, and 陳位彰. "The Design of Temperature Control Mechanism and Its Function on Nucleic Acid Quantification." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/fj79hp.
Повний текст джерела國立臺北科技大學
能源與冷凍空調工程系碩士班
96
This Real Time PCR machine allows the detection of DNA amplification through out the detection of the fluorescence labeling dye in the PCR mix during the early phase of this reaction. In addition, the concentration of target DNA fragment in the PCR mix before thermal cycling can be obtained from the time recorded history of the fluorescence intensity by integrating thermal cycler and fluorimeter. The Real-Time PCR machine has higher sensitivity and consumes less time than those of the traditional PCR machine. This study modified the temperature control mechanism developed by the previous studying results for the optimization of DNA amplification. A novel PID control program coded by Visual Basic was proposed to achieve optimized control of thermocycling of PCR. By 2% Agarose gel electrophoresis and UV absorption detection system, the yield products from this novel machine and the ABI 9700 PCR instrument were compared to verify the stability and reliability of the machine developed by our team. This results show that the temperature control scheme proposed in this study can be successfully applied in nucleic acid qualitative analysis. The 103 copies / ml initial copies sample has been successfully amplified. The 108 and 106 copies / ml samples were amplified successfully with high reproducibility. Regarding to the fluorescent detection system, a novel optical design was also proposed in this study. Because of the very weak fluorescent from the labeling dye attached on the DNA double strands, the fluorescent detection device itself should have very low dark current and the environmental light interferences should be avoided. The optical structure with black body painting surface was constructed to adopt cooled CCD based fluorescent detection devices to reduce the noise of fluorescent signal detection and isolate the sample under detection for high signal to noise ratio measurement of the fluorescence during PCR. The detection limit of this fluorescent detection system is 1 fmol Fluorescein. The signal can be resolved with the readings of the water. To summarize, this study has constructed a Real Time PCR machine for DNA amplification and quantification. In the future, we this instrument can be commercialized for a low cost and high accuracy DNA quantification solution.
Chen, Lin [Verfasser]. "Miniaturised nucleic acid analysis systems : purification, amplification and real-time detection / vorgelegt von Lin Chen." 2007. http://d-nb.info/997471891/34.
Повний текст джерелаHuang, Fu-Chun, and 黃富駿. "An Integrated Microfluidic System for Nucleic Acid Amplification, Electrophoresis Separation and On-line Optical Detection." Thesis, 2007. http://ndltd.ncl.edu.tw/handle/38805165998727799529.
Повний текст джерела國立成功大學
工程科學系碩博士班
96
This dissertation presents an integrated microfluidic chip capable of performing DNA/RNA (Deoxyribonucleic acid/Ribonucleic acid) amplification, electrokinetic sample injection and separation, and on-line optical detection of polymerase chain reaction (PCR) products in an automatic mode. In the device introduced here, DNA/RNA samples are first replicated using a micromachine-based PCR module or reverse transcription polymerase chain reaction (RT-PCR) module and then transported by a pneumatic micropump to a sample reservoir. The samples are subsequently driven electrokinetically into a microchannel, where they are separated electrophoretically and then detected optically using an optical fiber integrated into the device. The various modules of the integrated microfluidic chip are fabricated from cheap biocompatible materials, i.e. polydimethylsiloxane, polymethylmethacrylate and soda-lime glass. The functionality of the device is demonstrated through its successful application to the DNA-based bacterial detection of Streptococcus pneumoniae and the RNA-based detection of Dengue-2 virus. It is shown that the low thermal inertia of the PCR/RT-PCR modules reduces the sample and reagent consumption and shortens the reaction time. In order to further decrease the sample volume and to enhance disposability, a uniform temperature chip for PCR and capillary electrophoresis (CE) chip sealed with a polyethylene/thermoplastic elastomer (PE/TPE) film are developed. The uniform-temperature PCR chip was fabricated on soda-lime glass using MEMS technologies. A fence-type design of micro-heaters was used to improve the thermal uniformity of the reaction chamber. Experimental data show that the temperature distribution of the reaction chamber (diameter is 3 mm) is less than 1.0℃ in the PCR chamber at 52.0℃. The PE/TPE film packaged CE chip is performed at atmospheric pressure and room temperature, which is a fast, easy and reliable bonding method to form a sealed CE chip. The fabrication of polymethylmethacrylate (PMMA) and polycarbonate (PC) microfluidic channels is accomplished by using an injection-molding process, which could be mass-produced for commercial applications. In addition to microfluidic CE channels, three-dimensional reservoirs for storing bio-samples, and CE buffers are also formed during this injection-molding process. The functionality of the mass-produced CE chip is demonstrated through its successful separation of �珴-174/HaeⅢ DNA markers within 2 minutes. The integrated microfluidic device proposed in this study represents an important contribution to the fields of molecular biology, genetic analysis, infectious disease detection, and other biomedical applications.
Wang, Tzu-Chia, and 王子嘉. "Integrated Nucleic Acid Extraction, Amplification, and Detection on Paperfluidics for Diagnostics of Orchid Virus Disease." Thesis, 2019. http://ndltd.ncl.edu.tw/handle/sk5w8v.
Повний текст джерела國立臺灣大學
機械工程學研究所
107
In the study, we combined the extraction, amplification, and detection of nucleic acids in molecular diagnostics, and integrated them into paper-based microfluidics (paperfluidics) which was low-cost, lightweight, and portable. Paperfluidics reduced the complicated process and operation time compared to traditional molecular diagnostic technology and did not require precise instruments. This study developed point of care (POC) device that combined known technology and innovational methods, which could diagnose two major orchid viruses. First, the results showed that elliptic paperfluidics made by cellulose-based paper could capture and concentrate nucleic acids rapidly. The extraction process took only 10 minutes which was 12 times faster than using normal column purification. The second step was to amplify targeted virus genes by using recombinase polymerase amplification (RPA) at a constant temperature of 39°C. Compared with conventional amplification, polymerase chain reaction, RPA merely takes 30 minutes to amplify. Finally, using commercially available lateral flow strips for detection, we demonstrated the ability of the device to detect the two viruses CymMV and ORSV. Meanwhile, the results of the amplification reaction were analyzed by agarose gel electrophoresis. The best buffer was adding guanidine thiocyanate to extraction. Moreover, isothermal amplification and lateral flow assay integrated on the paper-based microfluidics achieved simple and rapid molecular diagnostics. The study successfully transformed molecular diagnostics into a portable paperfluidic device. We hope that the paper-based device will be used not only in the laboratory but also in the environments with limited resources such as farmland.
Huang, Hsuan-Shian, and 黃宣憲. "Nucleic acid detection, quantification and sequence analysis of canine distemper virus from clinical specimens." Thesis, 2005. http://ndltd.ncl.edu.tw/handle/95670168567838042013.
Повний текст джерела國立臺灣大學
獸醫學研究所
93
Canine distemper virus (CDV) infection in dogs can result in subclinical infection, gastrointestinal signs, and/or respiratory signs, frequently with central nervous system (CNS) involvement, high morbidity and mortality. Recently, the incidence of canine distemper (CD) both in unvaccinated and vaccinated dogs seemed increasing in Taiwan. In order to understand more about the current CDV infection in Taiwan, a rapid and sensitive diagnotic test for CD using a RT-PCR combined with nested PCR was applied to 440 dogs clinically suspected with CDV infection. 660 clinical specimens including CSF, whole blood, nasal swab, ocular swab, rectal swab and urine were collected from 440 dogs. The results showed that Nested PCR (37.7%, 166/440) increase 20.7% in the efficiency of the diagnosis than RT-PCR (17.0%, 75/489) (P<0.001). The seasonal distribution of the infection was mainly seen in late autumn to winter (P<0.001). Detection rate of conjunctival scraping (100.0%, 11/11) had a significant elevation compared with whole blood (64.4%, 38/59) (P<0.05). Quantification of the viral RNA concentration in clinical specimens from 27 cases by real-time PCR revealed that the highest concentration was as high as 1.8 x 1010 copies/mL. Different specimens from the same animal could vary up to 109 times. Nasal swab, ocular swab, oral swab and rectal swab had higher viral load, indicating that virus was shed in respiratory tract, digestive tract and urinary tract correlated to their clinical syndrome. Nasal swab was found to have the highest viral copies in cases with or without viremia. Therefore, the blood is not the best choice for clinical diagnosis. The result of H gene sequence analysis showed, that Taiwanese CDV is in the lineage of CDV Asia-1.
Ho, Ya-Lun, and 何亞倫. "Development of a Surface Acoustic Wave Based Micro-Droplet Control System and its Application of Nucleic Acid Amplification." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/43732841823831743871.
Повний текст джерела國立臺灣大學
應用力學研究所
99
In this thesis, an automatic micro-droplet control system applied to amplification of nucleic acid is accomplished by the combination of a surface acoustic wave (SAW) device, micro-heaters, micro-sensors of temperature, and a PI controller. The SAW device constituted of slanted finger interdigital transducers (SFITs) is utilized to actuate the micro-droplet by the acoustic streaming and to detect the micro-droplet by the frequency responses of the SAWs. With the development of PI controller, the micro-droplet can be manipulated automatically. In order to reduce the driving power and manipulate the mineral oil which is necessary for the reaction of the nucleic acid amplification, a perfluoroalkylsilane (PFAS) and tetraethoxysilane (TEOS) hydrophobic film is utilized for surface modification. Furthermore, with the PFAS/TEOS hydrophobic film, the cross-contamination can be prevented, and the micro-droplet control system is reusable for different DNAs and reagents. Utilizing the developed micro-droplet control system, the amplification of nucleic acid can be successfully achieved. The consumption of the biomedical reagent and the production cost of devices can be reduced, and the efficiency of reaction can be improved. The developed micro-droplet control system is suitable for various types of lab-on-a-chip system in biomedical fields.
Chen, Jiann-Mou, and 陳建謀. "A Rapid and Real-time Nucleic Acid Detection System by Fluorescence Resonance Energy Transfer and Isothermal RNA Amplification." Thesis, 2006. http://ndltd.ncl.edu.tw/handle/28118597521477734866.
Повний текст джерела國立臺灣大學
電機工程學研究所
94
In this thesis, we develop a detecting method with high accuracy and high sensitivity for specific RNA sequences in isothermal environment. We construct a low-cost and real-time system for detection specific RNA sequences by using this method. Isothermal RNA amplification is a powerful tool to amplify specific sequences of RNA. It can successfully amplify as low as 1 ng of total RNA in 90 minutes. Therefore, we integrate isothermal RNA amplification and Fluorescence Resonance Energy Transfer (FRET) methods to real-time detect fluorescent signals for specific RNA sequences. For increase specificity, two hybridization probes which can bind to the same gene are used in our experiments to increase the accuracy. Base on this study, a novel method with higher accuracy and sensitivity for detecting specific RNA sequence in isothermal environment can be achieved. The results shown that the FRET signal of TRIM28 and UCHL1 genes have been detected in 40-60 min. Therefore, we have successfully setup a new low-cost, real-time detection system, based on combined Isothermal RNA Amplification and FRET.
Goodrich, Terry T. "Ultrasensitive nucleic acid detection using surface plasmon resonance imaging measurements via a novel surface amplification process and microfluidic networks." 2004. http://www.library.wisc.edu/databases/connect/dissertations.html.
Повний текст джерелаCox, Philipp [Verfasser]. "Etablierung und Evaluation der NASBA (Nucleic-acid-sequence-based-amplification)-Technologie zum Nachweis von Aspergillus-RNA / vorgelegt von Philipp Cox." 2006. http://d-nb.info/97814404X/34.
Повний текст джерелаKosťun, Jan. "Využití molekulárně biologické metody One-Step Nucleic Acid Amplification (OSNA) při vyšetření sentinelových lymfatických uzlin u pacientek s karcinomem endometria." Doctoral thesis, 2018. http://www.nusl.cz/ntk/nusl-391393.
Повний текст джерелаOmar, S. V. (Shaheed Vally). "Innovative approaches to tuberculosis diagnosis with emphasis on nucleic acid amplification tests in a resource constrained high burden tuberculosis setting." Thesis, 2015. http://hdl.handle.net/2263/50754.
Повний текст джерелаThesis (PhD)--University of Pretoria, 2015.
tm2015
Medical Microbiology
PhD
Unrestricted