Using Real-Time Quantitative Reverse Transcriptase-Polymerase Chain Reaction to Screen Drug-Resistant Mycobacteria

• Main Authors: Wen-Cin Li, 李文欽
• Other Authors: Ming-Yi Liau
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/21821641777992182363

碩士 === 輔英科技大學 === 生物技術系碩士班 === 97 === Tuberculosis is a chronic infectious disease that has threatened human health for decades. Although it is curable and preventable, the emergence of drug-resistant Mycobateria tuberculosis strains have led to the low cure rate. Hence, effective drug treatments are very important. Traditional truberculosis drug susceptibility tests are specific but it takes 6-12 weeks to complete the culture and testing process and obtain the results. More infections and resistant strains may develop during this period. Therefore, the purpose of this study is to establish a rapid screening method using 16S rRNA and IS6110 real-time quantitative reverse-transcriptase polymerase chain reaction (real-time RT-PCR). The nonpathogenic Bacille Calmette-Gu�臆in vaccine (BCG) was used as the experimental materials. Our results showed, the detection range of 16S rRNA real-time RT-PCR is 1 x 107 to 1 x 102 BCG cells/mL. If the numbers of viable BCG are above 1 x 104 BCG cells/mL, after 14 days of incubation, the growth difference could be detected and the drug-resistant strains can be screened out. The detection range of IS6110 real-time RT-PCR is 1 x 107 to 1 x 105 BCG cells/mL. Similarily, if the numbers of viable BCG are above 1 x 104 BCG cell/mL, after 14 days of incubation, the growth difference could be detected and the drug-resistant strains can be screened out. However, the detection range of IS6110 real-time RT-PCR is narrow and the accuracy is low. Therefore, we suggested that 16S rRNA real-time RT-PCR system can be used in the rapid screening for drug-resistant M. tuberculosis, in contrast, IS6110 real-time RT-PCR system is not suitable for this purpose. In summary, the 16S rRNA real-time RT-PCR system established in this study can be combined with the traditional tuberculosis testing for the early screening of drug-resistant M. tubercolosis. It will be useful for the rapid diagnosis and reference for drug treatment.