磁性粒子和量子点快速筛查吹扫捕集气相色谱高灵敏特异鉴定分枝杆菌方法研究

Nowdays, Mycobacteria brings great threat to human health.However,the available methods can not meet the requirements for dtection.Hence,this project aims to develop a rapid,sensitive and specific method for the detection of mycobateria without culture..A lot of magnetic adsorbents and quantum dots showing selectivity to Mycobacteria are to be prepared and used to isolate Mycobateria from samples by forming complexes.This complexes can be isolated easily and Mycobateria can be detected rapidly and sensitively using fluorescence spectrometry based on luminescent properties of quantum dots. Besides,a simple device is to be constructed for the in-situ derivertization and purge-trap of the fatty acids in mycobacteria adsorbed into the complexes before analysis using gas chromatography. Using the in-situ derivertization and purgr-trap method combined with gas chromatography,mycobacteria in the complexes can be detected with high sensitivity and specificity after re-enrichment..In addition,the mechanism for the selective seperation of mycobacteria using the dye modified materials is to be explored based on the difference in adsorption behavior of various adsorbent, as well as the influence of experimental conditions on the adsorption behavior,combined with electron microscope.The value of the proposed method for the diagnosis of TB is to be evaluated after analysis being carried out on a large scale samples and compared with the commonly used clinical method..This research has great significance for the diagnosis of TB,which is one of the most important infectious diseases, and the proposed method is also suitable for the detection of all species of mycobateria other than Mycobateria tuberculosis,which is the pathogen of TB.

基于目前分枝杆菌感染不断增多、而现有检测手段不能满足要求的现状，本项目拟建立一套不经培养、快速、灵敏、特异地检测分枝杆菌的新方法。.制备对分枝杆菌有选择性的磁性粒子和量子点，利用磁性粒子、分枝杆菌、量子点形成三元复合体的发光性能，磁分离后用荧光法进行快速、灵敏初筛；研制简易的原位衍生化与吹扫-捕集和气相色谱联用装置，用于二次富集和分离分析三元复合体中分枝杆菌的脂肪酸组成，实现分枝杆菌的灵敏、特异检测。.通过研究不同吸附剂对分枝杆菌吸附行为的差异、考察各种实验条件对其吸附行为的影响、结合电镜等手段，对其分离机理进行探索；通过对大批量实际样品的测定，并与临床检测结果比较，评价该方法在TB诊断中的应用价值。.本课题的研究对于TB这一重要传染性疾病的诊断具有重要意义，所研究方法也适于分枝杆菌属其他细菌的检测。

针对本项目的研究内容，项目组分别从磁性粒子、量子点、吹扫捕集、分枝杆菌脂肪酸组成分析等方面进行了研究。制备出几种对分枝杆菌有选择性的磁性分离材料，并将其用于分枝杆菌和对照细菌的分离；制备出几种量子点，并研究了其与碱性品红、异烟肼的相互作用，考察了量子点的修饰及利用量子点进行分枝杆菌检测的可行性；设计了2种不同的吹扫捕集装置；建立了测定分枝杆菌中的脂肪酸，及依此进行分枝杆菌鉴定的方法。所取得的研究结果对于量子点、磁性粒子和吹扫捕集的应用以及分枝杆菌的检测都有一定借鉴意义。