DETECTION AND IDENTIFICATION OF MYCOBACTERIA IN CLINICAL SPECIMENS

临床标本中分枝杆菌的检测和鉴定

• 批准号: 6289476
• 负责人: Steven h FISCHER
• 金额: --
• 依托单位: CLINICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6289476
• 关键词: Mycobacterium; bacterial disease; communicable disease diagnosis; diagnosis design /evaluation; fluorescent dye /probe; polymerase chain reaction

Detection and identification of acid-fast bacilli of the genus Mycobacterium by conventional procedures requires growing the organisms from patient specimens and then testing the isolates for various phenotypic characteristics. These methods may require days to 1 or more months. The development of a few highly specific molecular probes for testing cultures growing acid-fast bacilli has greatly reduced the time to identification of some mycobacterial isolates. Recently, the polymerase chain reaction and isothermal nucleic acid amplification techniques have been used in assays that offer a high degree of specificity and reasonable sensitivity for detection of Mycobacterium tuberculosis in clinical samples. At present, there are no commercially available amplification assay systems that are capable of detecting multiple Mycobacterium species while excluding cross-reactive signals from other bacteria commonly present in clinical samples. We have initiated a new effort to investigate the usefulness of a pressure cycling technique to improve the efficiency of nucleic acid release from mycobacterial cells. The collaboration with Dr. Jang B. Rampal of Beckman Corporation to develop a better end point detection method for the 20 some most commonly encountered Mycobacterium species from human samples has continued. Experiments have been successfully performed with a method that simultaneously detects the presence or absence of nucleic acid amplifi-cation products from six common clinically isolated Mycobacterium species.

通过常规方法检测和鉴定分枝杆菌属的抗酸杆菌需要从患者标本中培养生物体，然后测试分离株的各种表型特征。这些方法可能需要几天到1个月或更长时间。一些高度特异性的分子探针用于检测抗酸杆菌的培养，大大缩短了鉴定某些分枝杆菌分离株的时间。最近，聚合酶链反应和等温核酸扩增技术已被用于检测临床样品中的结核分枝杆菌，提供了高度的特异性和合理的灵敏度。目前，还没有能够检测多个分枝杆菌物种同时排除来自临床样品中常见的其他细菌的交叉反应信号的市售扩增测定系统。我们已经开始了一项新的努力，以调查的有用性的压力循环技术，以提高效率的核酸释放从分枝杆菌细胞。与张博士B的合作。Beckman公司的Rampal开发了一种更好的终点检测方法，用于检测人类样本中最常见的20种分枝杆菌。用一种同时检测六种常见临床分离分枝杆菌核酸扩增产物的方法成功地进行了实验。