Establishment of a gold nanoparticle-based DNA detection platform for rapid diagnosis of Mycobacterium tuberculosis

• Main Authors: Yung Mao Chang, 張詠貿
• Other Authors: H. C. Lai
• Format: Others
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/22113831237661236278

碩士 === 長庚大學 === 醫學生物技術暨檢驗學系 === 99 === Tuberculosis is a worldwide common chronic disease mainly caused by Mycobacterium tuberculosis (MTB), especially in developing countries and countries under development. Therefore, to develop the diagnosis assays for accurate M. tuberculosis detection is urgent. The traditional diagnostic method is mainly achieved by bacteriological examination of clinical sputum samples from patients, including culture and acid stain smear screening; however, sputum culture is time-consuming. Furthermore, although acid-fast stain smear screening is much faster, the precision and alccuracy is not satisfactory. In this study, the molecular diagnostics, nano-science technology, and engineering principles were combined to try to develop new detecting platforms for M. tuberculosis detection. GNPs and LSPF technique for nucleic acid rapid detection of M. tuberculosis. It‘s very powerful that detection limit of this model is about 10.8 pg/ml of amplified DNA for ,. Also, it can do high-through put test for many specimens at the same time. The original nested PCR for PCR-ICR costed almost 2hrs, and the modified nested PCR costed about 1hr. Besides, 1st PCR and 2nd PCR of modified nested PCR can be work subsequently in the same eppendorf. What’s more, the efficiency of modified PCR-ICT is similar to original PCR-ICT. Also, we develop the GNP and immunoprecipiation assay in 96 wells plate for detection of amplified DNA of M. tuberculosis. The detection limit of this model is between 1-2 pmole(72.6-145.2 ng), and this result is not only similar to the GNP probe assay, but also it can do high-through put test for many specimens at the same time.