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Mechanistic studies of reactions at the single-molecule level using microfluidics with applications in molecular diagnostics

Citation

Sun, Bing (2014) Mechanistic studies of reactions at the single-molecule level using microfluidics with applications in molecular diagnostics. Dissertation (Ph.D.), California Institute of Technology. http://resolver.caltech.edu/CaltechTHESIS:05052014-142031176

Abstract

Motivated by needs in molecular diagnostics and advances in microfabrication, researchers started to seek help from microfluidic technology, as it provides approaches to achieve high throughput, high sensitivity, and high resolution. One strategy applied in microfluidics to fulfill such requirements is to convert continuous analog signal into digitalized signal. One most commonly used example for this conversion is digital PCR, where by counting the number of reacted compartments (triggered by the presence of the target entity) out of the total number of compartments, one could use Poisson statistics to calculate the amount of input target.

However, there are still problems to be solved and assumptions to be validated before the technology is widely employed. In this dissertation, the digital quantification strategy has been examined from two angles: efficiency and robustness. The former is a critical factor for ensuring the accuracy of absolute quantification methods, and the latter is the premise for such technology to be practically implemented in diagnosis beyond the laboratory. The two angles are further framed into a “fate” and “rate” determination scheme, where the influence of different parameters is attributed to fate determination step or rate determination step. In this discussion, microfluidic platforms have been used to understand reaction mechanism at single molecule level. Although the discussion raises more challenges for digital assay development, it brings the problem to the attention of the scientific community for the first time.

This dissertation also contributes towards developing POC test in limited resource settings. On one hand, it adds ease of access to the tests by incorporating massively producible, low cost plastic material and by integrating new features that allow instant result acquisition and result feedback. On the other hand, it explores new isothermal chemistry and new strategies to address important global health concerns such as cyctatin C quantification, HIV/HCV detection and treatment monitoring as well as HCV genotyping.

Item Type:Thesis (Dissertation (Ph.D.))
Subject Keywords:microfluidics; molecular diagnostics; isothermal amplification; viral load; genotyping; single molecule mechanism
Degree Grantor:California Institute of Technology
Division:Chemistry and Chemical Engineering
Major Option:Chemistry
Thesis Availability:Public (worldwide access)
Research Advisor(s):
  • Ismagilov, Rustem
Thesis Committee:
  • Dervan, Peter B. (chair)
  • Cai, Long
  • Pierce, Niles A.
  • Ismagilov, Rustem F.
Defense Date:2 May 2014
Non-Caltech Author Email:sunbing50 (AT) gmail.com
Record Number:CaltechTHESIS:05052014-142031176
Persistent URL:http://resolver.caltech.edu/CaltechTHESIS:05052014-142031176
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/ja4062882DOIArticle adapted for ch. 2
http://dx.doi.org/10.1021/ja2060116DOIArticle adapted for ch. 3
http://dx.doi.org/10.1021/ac3037206DOIArticle adapted for ch. 4
http://dx.doi.org/10.1021/ac4030413DOIArticle adapted for ch. 5
Default Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:8218
Collection:CaltechTHESIS
Deposited By: Bing Sun
Deposited On:12 May 2014 22:46
Last Modified:09 May 2016 17:36

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