Establish New Detection Of Tuberculosis For MicroRNAs As Target

Objective:Among a variety of worldwide infectious diseases at present, such as AIDS, Viral Hepatitis, Influenza, Tuberculosis and so on, tuberculosis(TB) is a common and serious chronic infectious disease that damages human health. Quick and efficient techniques for early diagnosis of tuberculosis are the premise of prevention and treatment. The microRNAs in the serum of TB patients and healthy people was used as object. The expression level of microRNAs in the serum of TB patients and healthy people was detected by real-time quantitative polymerase chain reaction(qRT-PCR) technique to find out the microRNAs molecules of stable differentially expressed and to establish new TB test methods using microRNAs as the target of detection in order to develop new diagnosis of TB. Methods:The expression levels of six kinds of microRNAs(miR-29 a, miR-93*,miR-320 b, miR-144*, miR-21, miR-155) which are highly correlated with TB in the serum of 50 TB patients and 50 healthy people were dected by qRT-PCR method to find stable differentially expressed microRNAs molecules which have statistical significance. According to screening results, the samples of stable differential expressed microRNAs were verified by qRT-PCR method. Meanwhile, the samples were detected by Enzyme-linked Immunosorbent Assay(ELISA). Then the two results were compared with each other to discuss whether microRNAs molecules could be applied to clinic and used as new biological markers to diagnosis TB, and to assess the clinical value of the new TB testing method aiming at microRNAs. Results:1.The expression level of miR-29 a in the serum of TB patients and that in the serum of healthy people had significant differences.The expression level of miR-320 b was significantly different. The other four microRNAs(miR-93*, miR-21, miR-155, miR-144 *) were not significantly different. 2.The fluorescence quantitative PCR reaction system of mi R-29 a was optimized. The optimal concentration of template was 5 times dilution retroviruses cDNA and the optimal primer concentration was 0.2μM. A new diagnosis technology of tuberculosis for miR-29 a as target was established. 3.The detection rate of tuberculosis was 94% which used mi R-29 a as target and positive detection rate was 89% with ELISA method. Conclusion:MiR-29 a was significantly different expressed in serum between tuberculosis group and controls. A kind of new diagnosis technology of tuberculosis which sees miR-29 a as target was established. This method is suitable for the TB diagnosis.