结核杆菌环介导等温扩增检测方法及间接ELISA检测方法的研究-预防兽医学专业论文.docxVIP

II II Abstract Tuberculosis that caused by Mycobacterium Tuberculosis is a chronic zoonosis. One-third of the world population is estimated to have Mycobacterium tuberculosis infection. Accurate and timely identifi cation of infected individuals is critical for treatment and control. For the detection of TB，tuberculin skin test (TST) is the only method recommended by OIE.Tubereulin is also called Purified Protein derivatives(PPD) which is a proteinmixture that has complex components.PPD includes many antigens shared with other Mycobacterium species besides specific antigens of M.bovis.So,the PPD skin test cannot avoid the cross reaction with other mycobaeterial.Moreover the method propably provides false Positive results when there is serious tubereulosis or immunosuppressive conditions exist. Moreover,the current diagnostic methods such as culture、smear、PCR and so on lack the desired sensitivity and specifi city, require sophisticated equipment and skilled workforce or take weeks to yield results. Therefore, this study developed two specific and sensitive diagnostic methods for TB. To develop a method of high efficiency, sensitivity and specificity to detect Mycobacterium tuberculosis and Mycobacterium bovis. Specifc primers were designed by targeting the esat-6 gene, gyrB gene and mtp40 gene. The results of LAMP detection were compared with the results of bacterium culture and PCR detection. The LAMP is a high efficient and specific method which can detect Mycobacterium tuberculosis and Mycobacterium bovis. The sensitivity of LAMP that can detect 7 copies/reaction was 100 times greater than PCR.Compared with three detection methods, it showed that the results of detection between LAMP and bacterium culture were 90.91% identity;the results of detection between LAMP and PCR were 100% identity. This new method might facilitate genetic analysis applied in clinical laboratory and field surveillance. Meanwhile, it also has some potentials for