| Mycobacterium tubercuLosis can secret a lot of protein in the process of growth,including the fusion protein CFP10 and ESAT6.This determination we used the aptamers detect a fusion protein CFP10 - ESAT6 which secreted by mycobacterium tuberculosis and we investgated thoroughly.The carboxyl of magnetic nanoparticles were as carrier which were modified by carboxyl, and we added the aptamers into the carrier. We used the methods of fluorescent assay, SDS gel electrophoresis and combination of complementary chain to confrim the proportion of carboxyl of magnetic nanoparticles and the aptamers.EVA Green embedded green fluorescent dyes in solutions of double-stranded DNA can be a strong fluorescence, and fluorescence intensity was weak in the single strand DNA.The green fluorescent dyes of EVA Green has strong fluorescent in solutions of double-stranded DNA and slightly in ssDNA. We added a balanced amount of bacteria solution into magnetic nanoparticles - dsDNA - Green fluorescent dye EVA Green system .If the aptamers and protein CFP10 - ESAT6 could Specific binding, the dsDNA can become into ssDNA and we can detect the specificity between the aptamers and the CFP10-ESAT6 by the changing of the fluorescence . As we all know, most of the protein have UV spectrums in 280 nanometers, so we use ultraviolet absorption method to do a rough quantitative the for the protein. Through this part, we wanted to achieve the goal of early diagnosis for tuberculosis.Polylactic acid(PLA) is a kind of biomedical material,which is widely used in drug loading and transport because the porous and biocompatibility. Rifampicin is one of the most widely used drugs to tuberculosis.In this experiment, we used the PLA as the carrier for the rifampicin, and then we tested the yield, capacity and embedding ratio were 63.56%, 13%, 20.35%. The later we added some aptamers into the system. We detected the fluorescence and ultraviolet absorption after fully shocked. The results suggest that the fluorescence is still strong and the losser of the rifampin is 18.46% between the compare to the standard curve of rifampicin. Through this part, we wanted to gain the end of treatment for tuberculosis and provided direction for the treatment of tuberculosis clinically. |
