Development of DNA probes for Mycobacterium tuberculosis and Mycobacteria other than Tuberculosis

• Main Authors: Lee, Mou-Shung, 李茂生
• Other Authors: Chin Tsung-Mei
• Format: Others
• Language: zh-TW
• Published: 1998
• Online Access: http://ndltd.ncl.edu.tw/handle/46798206639020728365

碩士 === 中國文化大學 === 應用化學研究所 === 86 === Mycobacteria infection are reemerging as important epidemic disease for the last several decades worldwide. The increased rate in mycobacterial infections has prompted the development of more rapid and efficient ways to detect and characterize mycobacteria spp., especially in the laboratory of clinical microbiology. Two procedures have long been adopted for the detection of acid-fast bacteria belonging to the family of Mycobacteriaceae. One is the direct microscopic examination of clinical specimen stained by Ziehl-Neelsen method, which is rapid, but is relatively insensitive and less specific. The other is a time-consuming culturing technique, which takes 4 to8 weeks of culturing time. We have developed a differential system based on the DNA-probing technology. The specificity and sensitivity of the DNA probes prepared from M. tuberculosis complex for detecting clinical samples are theoretically high. This non-isotope labeling DNA hybridization system should be suitable for laboratories that have a large number of specimens or in situation where the disposal of radioactive waste is a problem. The DNA probes, coupled to a non-radioactive tracer, e.g. digoxigenin-labeling and detection systems, has the potential to develop as a clinical assay for the rapid, specific, and sensitive identification of MTB or MOTT. Currently, we have isolated two mycobacterial DNA fragments: one is specific for the MTB complex and the other recognizes all other potentially pathogenic mycobacteria. These DNA fragments will be prepared as DNA probes to differentiate mycobacterial infection among the large amount of clinical specimens.