Kang Lin, Li Nan, Li Ping, Zhou Yang, Gao Shan, Gao Hongwei, Xin Wenwen, Wang Jinglin
1 State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, Beijing, People's Republic of China.
2 Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, People's Republic of China.
Eur J Mass Spectrom (Chichester). 2017 Apr;23(2):70-82. doi: 10.1177/1469066717699216. Epub 2017 Mar 24.
Salmonella can cause global foodborne illnesses in humans and many animals. The current diagnostic gold standard used for detecting Salmonella infection is microbiological culture followed by serological confirmation tests. However, these methods are complicated and time-consuming. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis offers some advantages in rapid identification, for example, simple and fast sample preparation, fast and automated measurement, and robust and reliable identification up to genus and species levels, possibly even to the strain level. In this study, we established a reference database for species identification using whole-cell MALDI-TOF MS; the database consisted of 12 obtained main spectra of the Salmonella culture collection strains belonged to seven serotypes. Eighty-two clinical isolates of Salmonella were identified using established database, and partial 16S rDNA gene sequencing and serological method were used as comparison. We found that MALDI-TOF mass spectrometry provided high accuracy in identification of Salmonella at species level but was limited to type or subtype Salmonella serovars. We also tried to find serovar-specific biomarkers and failed. Our study demonstrated that (a) MALDI-TOF MS was suitable for identification of Salmonella at species level with high accuracy and (b) that MALDI-TOF MS method presented in this study was not useful for serovar assignment of Salmonella currently, because of its low matching with serological method and (c) MALDI-TOF MS method presented in this study was not suitable to subtype S. typhimurium because of its low discriminatory ability.
沙门氏菌可导致人类和许多动物发生全球食源性疾病。目前用于检测沙门氏菌感染的诊断金标准是微生物培养,随后进行血清学确认试验。然而,这些方法复杂且耗时。基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)分析在快速鉴定方面具有一些优势,例如,样品制备简单快速、测量快速且自动化,以及在属和种水平甚至可能到菌株水平的鉴定稳健可靠。在本研究中,我们建立了一个使用全细胞MALDI-TOF MS进行物种鉴定的参考数据库;该数据库由属于七种血清型的沙门氏菌培养收集菌株的12个获得的主要光谱组成。使用建立的数据库对82株沙门氏菌临床分离株进行鉴定,并将部分16S rDNA基因测序和血清学方法用作比较。我们发现MALDI-TOF质谱在沙门氏菌物种水平的鉴定中提供了高精度,但仅限于沙门氏菌血清型的类型或亚型。我们还试图寻找血清型特异性生物标志物,但未成功。我们的研究表明:(a)MALDI-TOF MS适用于高精度鉴定沙门氏菌物种水平;(b)本研究中提出的MALDI-TOF MS方法目前对沙门氏菌血清型的分配无用,因为其与血清学方法的匹配度低;(c)本研究中提出的MALDI-TOF MS方法不适用于鼠伤寒沙门氏菌的亚型分析,因为其鉴别能力低。