Hurst G B, Doktycz M J, Vass A A, Buchanan M V
Chemical and Analytical Sciences Division, Oak Ridge National Laboratory, TN 37831-6365, USA.
Rapid Commun Mass Spectrom. 1996;10(3):377-82. doi: 10.1002/(SICI)1097-0231(199602)10:3<377::AID-RCM481>3.0.CO;2-X.
Accurate monitoring and identification of Legionella species, the causative agents of Legionnaires' and other diseases, in environmental water sources is an important public health issue. Traditional culture methods often lack the sensitivity and specificity that can be attained using the polymerase chain reaction (PCR) to amplify targeted regions of the bacterial genome. Matrix-assisted laser desorption/ionization combined with time-of-flight (MALDI-TOF) mass spectrometry is shown to be useful for detection of 108- and 168-base PCR products specific to Legionella. A rapid purification aimed at removal of salts and unreacted primers is demonstrated. The addition of a synthetic DNA 20-mer to the MALDI sample facilitates aiming the laser at a favorable spot on the sample probe from which the PCR products can be detected.
准确监测和鉴定军团菌属（军团病及其他疾病的病原体）在环境水源中的存在是一个重要的公共卫生问题。传统培养方法往往缺乏使用聚合酶链反应（PCR）扩增细菌基因组靶向区域所能达到的灵敏度和特异性。基质辅助激光解吸/电离与飞行时间（MALDI-TOF）质谱联用已被证明可用于检测军团菌特有的108和168碱基PCR产物。展示了一种旨在去除盐分和未反应引物的快速纯化方法。向MALDI样品中添加一条20聚体合成DNA有助于将激光对准样品探头上能检测到PCR产物的有利位置。
