The Applications of Microarray－Diagnosis of Pathogenic Microorganisms

• Main Authors: Shang-Kai Hung, 洪尚楷
• Other Authors: Ruey-Shyang Chen
• Format: Others
• Language: zh-TW
• Published: 2002
• Online Access: http://ndltd.ncl.edu.tw/handle/86981952065529803063

碩士 === 國立嘉義大學 === 生物科技研究所 === 90 === Microarray is a novel and powerful tool for genomic studies. Based on its high capacity and specificity, the main objective of the present study is to develop a biochip for the diagnosis of pathogenic microorganisms. 71 EBV genomic DNA fragments of 1-3 kb in length were amplified by PCR using specific primers designed from EBV genome. These amplified products were spotted to make the EBV-chip by a microarrayer. To test the EBV-chip, mRNA extracted from EBV-infected P3HR1 cell line were transcribed into cDNA and labeled with biotin-16-dUTP. After hybridization and color development, the signals on EBV-chip were scanned and analyzed. The distribution of EBV in different tumor tissues were further assayed by the EBV-chip. On the other side, the Mycochip was made to detect and differentiate plant pathogenic fungi in the present study. Using the universal primers, ribosomal DNA (rDNA) including internal transcribed spacer (ITS) were amplified from 48 fungal isolates, and were spotted by a microarrayer. DNA extracted from tested fungal isolates and diseased plant tissues were PCR-amplified, labeled, and hybridized with the Mycochip. The results showed that different pathogenic fungi could be simultaneously detected and classified in diseased plant tissues. The Mycochip could also be used to differentiate fungal species form various sources. Based on the present study, the biochip has shown its great potential for the diagnosis of pathogenic microorganisms. Microarray technology also possessed several advantages over the current diagnostic methods, such as PCR and in situ hybridization, that were discussed in this study.