Rapid Diagnosis of Mycobacterium tuberculosis and Drug Resistance by SPR-Based Method (I)

Tuberculosis (TB) remains one of the most life-threatening infectious diseases and causes two million deaths per year and eight million new case of TB infection. Mycobacterium tuberculosis is the causative agent of TB and remains one of the leading causes of respiratory infections and has posed critical threats to public health. Although the control of tuberculosis is significantly effective in recent years in Taiwan, Taiwan is still the high prevalence areas. Therefore, prevention of infection and surveillance of incomplete treatment are major factors in affecting the prevention and control of tuberculosis in Taiwan. The major pathogen for tuberculosis among Mycobacterium tuberculosis complex (MTBC) is M. tuberculosis. The major syndrome is pulmonary tuberculosis and the most often clinical specimens are sputum. Traditional methods of tuberculosis diagnosis are culture and acid-fast stain. It is time-consuming and the sensitivity and specificity is relatively low. Rapid detection of TB infection is important to the control of this disease. Hence, we are going to develop a rapid TB detection system based on our developmental foundation with the actual needs of the national policy to shorten the test time, savings and to increase sensitivity and specificity. This system includes (I) rapid nucleic acid detection system of M. tuberculosis, and (II) rapid diagnosis system of TB drug resistance. In addition to IS6110, Gene sequence alignment reports showed that M. tuberculosis has unique DNA sequences such as RD1 to RD16. In this proposal, clinical samples will be tested by the SPR chip system for rapid diagnosis. By using IS6110 and RD9 region as the target for DNA amplification technology, Loop Mediated Isothermal Amplification (LAMP), the sensitivity and specificity of rapid diagnosis of TB would be improved. In the development of TB drug resistance detection system, we will also use known drug resistance mutation gene of M. tuberculosis. The variations of drug-resistant genes of M. tuberculosis will be used as target in diagnosis of drug resistance mutations of M. tuberculosis in real time with continuous improvement of reagent design. First of all, four drugs will be tested for nucleic acid diagnosis, and the time-consuming would be significantly decreased. This scheme is funded through the results and experience to develop novel, accurate, affordable and easy to use rapid detection system of TB, the SPR-based rapid diagnosis of TB and TB drug resistance will be able to quickly and accurately diagnose source of TB infection, to achieve the benefits of early diagnosis and early treatment. If our excellent research and development team could tie in with the promotion of National Science Council Grant, the technique of SPR-LAMP -based detection system would significant decrease the cost and achieve the dual purpose of prevention and treatment of TB infection.

Project ID:PC10006-0031
External Project ID:NSC100-2325-B182-005