Mycobacteria in metalworking fluids

金属加工液中的分枝杆菌

• 批准号: 7256941
• 负责人: Jagjit S Yadav
• 金额: $ 26.59万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-30 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7256941
• 关键词: Mycobacteria; metalworking; fluids

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.

非结核分枝杆菌（NTM）经常定居于现代金属加工液（MWF），为 越来越多地与职业呼吸道疾病有关，例如高敏性肺炎（HP） 在机器工人中。他们的早期检测/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PCR的协议的制定 水基MWF中非培养的非依赖性检测和定量，并与分离和 MWF的几种NTM菌株的基因型鉴定，这些NTM菌株属于两个物种。 免疫原和切氏菌。我们提出的未来目标是开发下一代简化 基于DNA的方法可以在内部实验室中进行调整并研究相对杀菌剂 这些MWF分离的NTM菌株（基因型）的敏感性和抗原潜力。具体目的是 （1）。开发有效的NTM恢复和简单基于DNA的检测和定量的协议 来自不同MWF类型的NTM； （2）。确定可用和新的相对生物剂易感性 通过优化和使用高吞吐量协议，来自MWF的NTM菌株； （3）。研究抗原 可用和HP连接的NTM菌株的特征。免疫磁分离 分枝杆菌特异性多克隆抗体将开发用于选择性细胞恢复以改进 NTM检测和文化隔离。我们当前的PCR协议将用于比色检测 NTM。将开发基于荧光原位杂交（FISH）的新方案用于显微镜 - 辅助培养的检测和MWF特异性分枝杆菌物种的定量。高 基于细胞内ATP的吞吐量协议将被优化并用于测量biocide 敏感性。 MWF降低菌株/基因型的相对抗原潜力将基于 关于HP患者或MWF暴露工人的敏化血细胞中细胞免疫反应。 M. 免疫原子多克隆抗体或HP患者血清将作为探针识别推定 鉴定抗原菌株的细胞蛋白裂解物和培养滤液中的蛋白抗原。结果 信息将通过为分枝杆菌提供一组实用方法来帮助NORA的目标 污染和暴露评估以及应变特征数据库，以促进发展 与MWF暴露有关的干预策略。