Madonna A J, Basile F, Furlong E, Voorhees K J
Department of Chemistry and Geochemistry, Colorado School of Mines, Golden, CO 80401, USA.
Rapid Commun Mass Spectrom. 2001;15(13):1068-74. doi: 10.1002/rcm.344.
A rapid method for identifying specific bacteria from complex biological mixtures using immunomagnetic separation coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry has been developed. The technique employs commercially available magnetic beads coated with polycolonal antibodies raised against specific bacteria and whole cell analysis by MALDI-MS. A suspension of a bacterial mixture is mixed with the immunomagnetic beads specific for the target microorganism. After a short incubation period (20 mins) the bacteria captured by the beads are washed, resuspended in deionized H(2)O and directly applied onto a MALDI probe. Liquid suspensions containing bacterial mixtures can be screened within 1 h total analysis time. Positive tests result in the production of a fingerprint mass spectrum primarily consisting of protein biomarkers characteristic of the targeted microorganism. Using this procedure, Salmonella choleraesuis was isolated and detected from standard bacterial mixtures and spiked samples of river water, human urine, and chicken blood.
已开发出一种快速方法,用于从复杂生物混合物中鉴定特定细菌,该方法使用免疫磁珠分离结合基质辅助激光解吸/电离飞行时间质谱。该技术采用涂有针对特定细菌的多克隆抗体的市售磁珠,并通过基质辅助激光解吸/电离飞行时间质谱进行全细胞分析。将细菌混合物的悬浮液与针对目标微生物的免疫磁珠混合。短时间孵育(20分钟)后,将被磁珠捕获的细菌洗涤,重悬于去离子水中,然后直接涂覆到基质辅助激光解吸/电离飞行时间质谱探头上。含有细菌混合物的液体悬浮液可在1小时的总分析时间内进行筛选。阳性测试会产生主要由目标微生物特有的蛋白质生物标志物组成的指纹质谱。使用该程序,从标准细菌混合物以及河水、人尿和鸡血的加标样品中分离并检测出猪霍乱沙门氏菌。