Mycobacteria in metalworking fluids

非结核分枝杆菌(NTM)经常在现代金属加工液(MWF)中定植，并且 越来越多地与职业呼吸系统疾病有关，如过敏性肺炎(HP) 在机器工人身上。它们的早期发现/恢复和特征将有助于采取先发制人的措施 并制定干预策略。我们的第一笔资金导致了基于聚合酶链式反应的方案的开发 与培养无关的水基MWF中NTM的检测和定量，结合分离和 对MWF的几株NTM菌株进行了基因分型鉴定。 免疫原菌和龟分枝杆菌。我们提出的未来目标是开发下一代简化的 基于DNA的方法，可以在内部实验室中采用，并调查相关的杀生剂 这些MWF分离的NTM菌株(基因型)的敏感性和抗原性。具体目标是 (1)。开发有效的NTM恢复和简单的基于DNA的检测和量化的方案 来自不同MWF类型的NTM；以确定现有的和新的杀菌剂的相对敏感性 通过优化和使用高通量方案从MWF中分离出NTM菌株；调查抗原性 现有的与幽门螺杆菌相关的NTM菌株的特征。免疫磁选分离技术 分枝杆菌特异性多克隆抗体将被开发用于选择性细胞回收 NTM检测和培养分离。我们目前的聚合酶链式反应方法将适用于比色法检测 新台币。一种基于荧光原位杂交(FISH)的新方法将被开发用于显微镜- 不依赖于培养的辅助检测和MWF特定分枝杆菌种类的量化。一次高潮 基于胞内ATP的吞吐量方案将被优化并用于杀菌剂的检测 敏感度。MWF流行株/基因型的相对抗原性将根据以下因素确定 Hp患者或MWF接触工人致敏血细胞免疫反应的研究M. 免疫原性多克隆抗体或幽门螺杆菌患者血清将被用作探针来识别推定的 鉴定的抗原菌株的细胞蛋白裂解物和培养滤液中的蛋白质抗原。由此产生的 信息将通过为分枝杆菌提供一套实用的方法来帮助Nora的目标 污染和暴露评估和菌株特征数据库，以促进发展 与MWF暴露相关的干预策略。

Non-tuberculous mycobacteria (NTM) frequently colonize the modem metalworking fluids (MWFs) and are increasingly being implicated in occupational respiratory illnesses such as hypersensitivity pneumonitis (HP) in machine workers. Their early detection/recovery and characterization will help take pre-emptive measures and develop intervention strategies. Our first funding has led to the development of PCR based protocols for culture-independent detection and quantification of NTM in water-based MWF, coupled with isolation and genotypic identification of several NTM strains from MWF, which belonged to two species M. immunogenum and M. chelonae. Our proposed future goals are to develop next generation of simplified DNA-based approaches that can be adapted in in-house laboratories and to investigate the relative biocide susceptibility and antigenic potential of these MWF isolated NTM strains (genotypes). The specific aims are (1). to develop protocols for efficient NTM recovery and simple DNA-based detection and quantification of NTM from different MWF types; (2). to determine the relative biocide susceptibility of the available and new NTM strains from MWF by optimization and use of a high throughput protocol; (3). to investigate antigenic characteristics of the available and HP-linked NTM strains. Immunomagnetic separation using Mycobacterium-specific polyclonal antibodies will be developed for selective cell recovery for improved NTM detection and cultural isolation. Our current PCR protocol will be adapted for colorimetric detection of NTM. A new protocol based on Fluorescence in-situ hybridization (FISH) will be developed for microscope- assisted culture-independent detection and quantification of the MWF-specific mycobacterial species. A high throughput protocol based on intracellular ATP will be optimized and used for measurement ofbiocide susceptibility. Relative antigenic potential of the MWF-prevalent strains/genotypes will be determined based on cellular immune response in the sensitized blood cells from HP patients or MWF exposed workers. M. immunogenum polyclonal antibodies or HP patient sera will be applied as probes to identify the putative protein antigens in the cell protein lysates and culture filtrates of the identified antigenic strain. The resulting information will help NORA's objectives by providing a set of practical methods for mycobacterial contamination- and exposure- assessment and a strain characteristics database, for facilitating development of intervention strategies related to MWF exposures.