| Mycobacterium tuberculosis(M.tuberculosis) may invade each organs of whole body, but most common is lung infection. As an important infectious diseases tuberculosis is also threat to human health seriously. With 9 million new M.tuberculosis infections worldwide every year, and showed overall upward trend.The emergence of multi-drug resistance and human immunodeficiency virus infection has caused explosive increases in TB incidence. Therefore, early diagnosis and treatment of TB is one of the important measures to effectively control the epidemic.At the same time, M.tuberculosis can lead to asymptomatic latent tuberculosis infection and active tuberculosis. Latent tuberculosis infection(LTBI) is a special state of the disease which have not the incidence after the host infected by M.tuberculosis. However, most of LTBI will eventually converted into active TB. As the same, it is very necessary to diagnosis latent tuberculosis infection timely.In recent years, specific antigens and surface membrane protein secreted by M.tuberculosis has became a hot research points for early diagnosis of tuberculosis.Glycolipids substances of M.tuberculosis cell wall has genus-specific and with depth study we can know that early culture filtrate protein CFP10-ESAT6 has good immunogenicity and the presence of the gene encoding the RD1 region which exists only few pathogenic mycobacteria and M.tuberculosis complex, absent from BCG and non-pathogenic mycobacteria.Therefore, distinguish between pathogenic and non-pathogenic of M.tuberculosis has more research value. This paper, aims to establish a new method for early diagnosis of M.tuberculosis. The main work is as follows:1. According to the literature, CFP10-ESAT6 has a higher sensitivity and specificity compared to its single antigen. Its characteristics are able to distinguish between natural infection of M.tuberculosis, BCG vaccination and non-tuberculous mycobacterial infection. Therefore, it can be used as an antigen-specific for diagnosis of tuberculosis. In this paper, gold nanoparticles prepared by chemical reduction.Chlorine acid as a raw material, sodium citrate as reducing agent to prepare a particle size of approximately 17 nm of gold nanoparticles. It has been characterized by uv-vis absorption spectroscopy and transmission electron microscopy. The results showed that the gold nanoparticles has a strong absorption peak at 520 nm, and the particle size of approximately 17 nm. In this article aptamer molecule labeled by nano-goldthrough thiol self-assembly, compared with the stability of aptamer after the modification and nano gold sol by UV-visible absorption spectrum.In this paper, we successfully constructed the IDE-SPQC sensor combined with nano-gold and aptamer technology detect the protein CFP10-ESAT6 for early tuberculosis diagnosis rapidly.The results showed that M.tuberculosis can be detected within 96.3 hours by our sensor. And the detection finished in 15 min. The method is more simple, rapid,sensitive compared to common clinical immunoassay which is expected to become a new method for early clinical diagnosis of M.tuberculosis.2. LTBI is not yet developed a host infected with TB after active tuberculosis clinical manifestations. In low immunity likely to develop active tuberculosis and lead to corresponding clinical symptoms that becomes a new source of infection. During the initial infection with M.tuberculosis, the immune system is activated to produce memory T lymphocytes and the effector of T lymphocytes for M.tuberculosis, when second infected by M.tuberculosis antigen, effector T lymphocytes cells are activated,secrete large amounts of IFN-?. By detect levels of quantitative of the IFN-? can diagnose tuberculosis infection situation. Series piezoelectric sensor has many advantages have been widely used microbial sensing and analysis which due to the solution and the electrode surface conductivity and dielectric constant conductance change cause the frequency shift response. Interdigital microelectrode array consists of a series of parallel, interdigitated electrodes microstrip and small changes can be sensitively detected on the electrical parameter between electrodes. In this paper,IDE-MSPQC sensors combined aptamer and conductive polymer Co(phen)33+respond to IFN-? successfully. This high sensitivity, high specificity new method of IFN-? is expected to improve practical ability recessive clinical diagnosis of tuberculosis. |
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