摘要
目的建立临床常见三种葡萄球菌的蛋白指纹图谱,为其快速诊断奠定基础。方法收集临床常见葡萄球菌包括金黄色葡萄球菌、表皮葡萄球菌及溶血性葡萄球菌,利用16S rDNA测序技术对其进行鉴定;表面增强激光解吸电离飞行时间质谱(SELDI-TOF MS)技术检测其蛋白表达,采用Ciphergen Protein chip软件自动采集数据。评价该检测方法的重复性及细菌蛋白表达的稳定性。结果在分子量3 000～20 000 Da范围内,3种葡萄球菌有各自特异的蛋白指纹图谱;SELDI-TOF MS检测细菌蛋白的孔间重复性和芯片间重复性好,同一株细菌4次重复的蛋白指纹图谱相似;同种细菌不同株之间蛋白指纹图谱表达稳定,共有蛋白峰分子量变异系数≤0.5%。结论在分子量3 000～20 000 Da范围内,各细菌蛋白指纹图谱差异明显,为细菌的快速鉴定提供了新方法。
Objective To establish protein fingerprints of clinically common staphylococci to provide basis for rapid identification of the bacteria. Methods 60 clinical isolates including Staphylococcus aureus, Staphylococcus epidermidis and Staphylococcus haemolyticus were collected and studied. 16S rDNA sequencing was used to identify all of the strains as a gold standard. Then the isolates were analyzed by surface enhanced laser desorption/ionization time-of-flight mass spectra （SELDI-TOF MS） and the data were collected by Ciphergen ProteinChip software. The reproducibility of SELDI-TOF MS and the stability of the protein expression of bacteria from each species were analyzed based on the protein fingerprints. Results The protein profile of the same bacterium had good reproducibility. Bacteria of different species had various protein fingerprints from 3 kD to 20 kD while the same species had similar protein fingerprints and the CV of molecular weight of main protein biomarkers was less than 0.5%. Conclusion There are various characteristic protein fingerprints in bacteria of different species, which pro- vides a new approach for rapid identification of bacteria.
出处
《中国微生态学杂志》
CAS
CSCD
2012年第11期1000-1002,共3页
Chinese Journal of Microecology
