Construction And Application Of Novel Siderophore Sensor For Rapid Detection Of Mycobacterium Tuberculosis

Tuberculosis caused the highest mortality in worldwide, there will be about1.8million people die of tuberculosis every year. Although in most developed countries,the probability of occurrence of TB is very low, but in the economically backwardcountries, especially African countries and Southeast Asian countries, there is still ahigh prevalence of TB. The growth of M. tuberculosis growth very slow, resulting inthe clinical diagnosis being a longer period than other pathogens. And because ofirrational drug use during treatment, leading to the emergence of resistant strains oftuberculosis. Therefore, there is an urgent need for an effective method for rapiddiagnosis of tuberculosis and tuberculosis pathogenicity. This paper aims to establisha new method to detect tuberculosis, to achieve rapid diagnosis of tuberculosis. Themain work is as follows:Siderophore is a potential biomarker for pathogen diagnosis. In this study, asiderophore impedance sensor was constructed based on the formation ofsiderophore-hemin complex which generated during the interaction between the two.The siderophore probe was prepared by modification of hemin on interdigitated arraymicroelectrode. When siderophore existed in sample, it would react with hemin andformed siderophore-hemin complex covering the electrode surface instead of heminonly. As the conductivity of hemin was higher than that of siderophore, the formationof siderophore-hemin complex led to decrease of conductivity on electrode surfacewhich could be measured by electrochemical impedance spectroscopy. Compared withthe common siderophore detection methods including chrome azurol S-shuttlesolution and ferric perchlorate assay, this method is more simple, rapid and sensitive.It could be applied to clinical pathogen detection and drug test.Construction and applied to the detection of Mycobacterium tuberculosiscontaining iron body of piezoelectric sensors.Mycobacterium tuberculosis secretedsiderophore which can be combined with the hemin. In this study, a siderophorepiezoelectric sensor was constructed based on the formation of siderophore-hemincomplex which generated during the interaction between the two. The siderophoreprobe was prepared by modification of hemin on interdigitated array microelectrode.When siderophore existed in sample, it would react with hemin and formedsiderophore-hemin complex covering the electrode surface instead of hemin only. As the conductivity of hemin was higher than that of siderophore, the formation ofsiderophore-hemin complex led to decrease of conductivity on electrode surfacewhich could be measured by an series piezoelectric quartz. The conductivity changeon the electrode surface is in accordance with the frequency shift of SPQC, thus thedetection of Mycobacterium tuberculosis constructed by realtime detection of thesiderophore secretion.A new interdigitated electrode-piezoelectric quartz crystal senor for rapiddetection of Mycobacterium was constructed. Mycobacterium Smegmatis, anon-pathogenic bacteria, was used as a representative model in this experiment, whichpropose similar structural features with Mycobacterium tuberculosis. In this study, acommon siderophore, deferential was modified on interdigitated array microelectrode.There is a specific recognition between the siderophore and the siderophore receptoron the cell wall of bacteria. In the presence of bacteria, the siderophore modified onelectrode could harvest bacteria by combining with its receptor site, therefore thebacteria could be absorbed on the surface of IDAM, which caused dramatic impedancechange on the electrode surface. The SPQC was employed to construct a real-timedetection of the impedance change on the electrode and response to it by acorresponding change of frequency. This sensor achieved a rapid, sensitive andsimple detection of Mycobacterium tuberculosis.