000			04906nas a2200457 a 4500
005			20260818160037.0
008			080821s2008 th uu|m rtt 0| a1eng d
035			_a.b12013602
099		9	_aAIT Thesis no.EV-08-6
100	1		_aOrtinero, Cesar
245	1	0	_aNanoparticle-based development of selected innovative nucleic acid-related microbiological methodologies
260			_aPathum Thani, Thailand : _bAsian Institute of Technology, _c2008
300			_a65 leaves : _bill. + _e1 online resource
490	1		_aThesis ; _vno. EV-08-6
500			_aA thesis submitted in partial fulfillment of the requirements for the degree of Master of Science in Environmental Engineering and Management Inter-University Program on Environmental Toxicology, Technology and Management
502			_aThesis (M.Sc.) - Asian Institute of Technology - Chulabhorn Research Institute - Mahidol University, 2008
520			_aFluorescence in situ hybridization (FISH) and amplification by polymerase chain reaction (PCR) are two of the most commonly applied molecular microbiological methods. A major limitation of FISH is its dependence on organic fluorophores whose susceptibility to light-induced degradation forces researchers to work in the dark. Frequent inefficiency and non-specificity are the major drawbacks of PCR. The main goal of this study was to develop alternative methodologies overcoming shortcomings of these two methods through an application of nanoparticles. An innovative FISH methodology with superior photostability and biological compatibility was developed through the use of the chitosan-capped Mn-doped ZnS quantum dots (QDs) as fluorescence label for the phylogenetic oligonucleotide probes. Stringent hybridization between the QD-labeled oligonucleotide probes and target rRNA site in E. coli was carried out in a formamide-free buffer under high temperature (55-65°C) incubation. Highly luminescent, the QDs enabled the efficient detection by epifluorescence microscopy of E. coli cells which were 75% brighter than the background fluorescence. Extremely photostable, the QDs eliminated the need to work in the dark and the use of antifade reagents. In its current form, the QD-FISH methodology is somewhat limited by the interference from autofluorescence, which requires additional research. Meanwhile, image processing and analysis can help overcome this limitation and demonstrate the effectiveness of the QD-FISH technique in detecting E. coli from a mixture of bacteria. Case-specific improvement of the efficiency and specificity of amplification of nucleic acids was attained through the addition of either 20-nm silica (SiO₂) or 15-nm gold (Au) nanoparticles in PCR systems (--10¹² particles in 25-uL PCR mixture). One of the proposed mechanisms for the enhancement of nucleic acid amplification by Au nanoparticles revolves around the excellent heat conductivity of Au nanoparticles which leads to efficient thermal cycling in PCR. In this study, similar patterns of inhibition and enhancement of nucleic acid amplification were observed in PCR systems supplemented with either Au or SiO₂ nanoparticles, despite the difference in ability of these nanoparticles to conduct heat. Thus, the role of thermal conductivity on nanoparticle-PCR, though important, only provides a partial explanation and the interaction between the nanoparticles and the components of the PCR system appears to play a more significant role in the nanoparticle-based enhancement of nucleic acid amplification
650		0	_aNanoparticles
650		0	_aNucleic acids
700	1		_aShippin, Oleg V., _eChairperson
700	1		_aDutta, Joydeep, _eExamination Committee
700	1		_aVisvanathan, C., _eExamination Committee
700	0		_aSkorn Mongkolsuk, _eExamination Committee
710	2		_aIFP, _eScholarship donor
710	2		_aFord Foundation, _eScholarship donor
710	2		_aAsian Institute of Technology Fellowship, _eScholarship donor
810	2		_aAsian Institute of Technology. _tThesis ; _vno. EV-08-6
856			_3Full-Text _uhttp://203.159.5.9/ait-thesis/detail.php?q=B02438
907			_a.b12013602 _bmnait _cu
902			_a240401
998			_b0 _c080821 _dm _ea _fu _g0
945			_lmnait
945			_lmnait
945			_lmnarc
942			_c20
942			_c40
909			_aBarcode : 30050120736490 _bCREATED : 2005-12-25 _cRECORD # : i12525832 _dLPATRON : 0 _eLCHKIN : - _f# RENEWALS : 0 _g# OVERDUE : 0 _hIUSE3 : 1 _iTOT CHKOUT : 0 _jTOT RENEW : 0
909			_aBarcode : 30050120736466 _bCREATED : 2007-03-24 _cRECORD # : i12570199 _dLPATRON : 1019522 _eLCHKIN : 2008-12-16 _f# RENEWALS : 0 _g# OVERDUE : 0 _hIUSE3 : 0 _iTOT CHKOUT : 2 _jTOT RENEW : 0
909			_aBarcode : 30050160049242 _bCREATED : 2016-05-31 _cRECORD # : i13005418 _dLPATRON : 0 _eLCHKIN : - _f# RENEWALS : 0 _g# OVERDUE : 0 _hIUSE3 : 0 _iTOT CHKOUT : 0 _jTOT RENEW : 0
999			_c79537 _d79537