Paauw Armand, Trip Hein, Niemcewicz Marcin, Sellek Ricela, Heng Jonathan Me, Mars-Groenendijk Roos H, de Jong Ad L, Majchrzykiewicz-Koehorst Joanna A, Olsen Jaran S, Tsivtsivadze Evgeni
Department of CBRN Protection, TNO, P,O, Box 45, Rijswijk 2280 AA, The Netherlands.
BMC Microbiol. 2014 Jun 18;14:158. doi: 10.1186/1471-2180-14-158.
Cholera is an acute diarrheal disease caused by Vibrio cholerae. Outbreaks are caused by a genetically homogenous group of strains from serogroup O1 or O139 that are able to produce the cholera toxin. Rapid detection and identification of these epidemic strains is essential for an effective response to cholera outbreaks.
The use of ferulic acid as a matrix in a new MALDI-TOF MS assay increased the measurable mass range of existing MALDI-TOF MS protocols for bacterial identification. The assay enabled rapid discrimination between epidemic V. cholerae O1/O139 strains and other less pathogenic V. cholerae strains. OmpU, an outer membrane protein whose amino acid sequence is highly conserved among epidemic strains of V. cholerae, appeared as a discriminatory marker in the novel MALDI-TOF MS assay.
The extended mass range of MALDI-TOF MS measurements obtained by using ferulic acid improved the screening for biomarkers in complex protein mixtures. Differences in the mass of abundant homologous proteins due to variation in amino acid sequences can rapidly be examined in multiple samples. Here, a rapid MALDI-TOF MS assay was developed that could discriminate between epidemic O1/O139 strains and other less pathogenic V. cholerae strains based on differences in mass of the OmpU protein. It appeared that the amino acid sequence of OmpU from epidemic V. cholerae O1/O139 strains is unique and highly conserved.
霍乱是由霍乱弧菌引起的急性腹泻病。疫情爆发是由来自血清群O1或O139的一组基因同质菌株引起的，这些菌株能够产生霍乱毒素。快速检测和鉴定这些流行菌株对于有效应对霍乱疫情至关重要。
在一种新的基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）检测中使用阿魏酸作为基质，扩大了现有用于细菌鉴定的MALDI-TOF MS检测方法的可测量质量范围。该检测方法能够快速区分霍乱弧菌O1/O139流行菌株和其他致病性较低的霍乱弧菌菌株。外膜蛋白OmpU在霍乱弧菌流行菌株中氨基酸序列高度保守，在新型MALDI-TOF MS检测中作为一种鉴别标志物出现。
使用阿魏酸获得的MALDI-TOF MS测量扩展质量范围改善了对复杂蛋白质混合物中生物标志物的筛选。由于氨基酸序列变化导致的丰富同源蛋白质质量差异可在多个样品中快速检测。在此，开发了一种快速MALDI-TOF MS检测方法，该方法可根据OmpU蛋白质量差异区分霍乱弧菌O1/O139流行菌株和其他致病性较低的霍乱弧菌菌株。霍乱弧菌O1/O139流行菌株的OmpU氨基酸序列似乎是独特且高度保守的。
