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目的:利用表面增强激光解析电离飞行时间质谱建立甲型副伤寒沙门氏菌蛋白指纹图谱。方法:收集临床中分离获得的甲型副伤寒沙门氏菌36株,同时采集对照菌96株,对所收集的细菌利用16S rDNA测序技术进行分子生物学验证鉴定。利用表面增强激光解析电离飞行时间质谱检测细菌蛋白,ProteinChip和Biomarker Wizard软件自动采集数据,将结果用BioMarker Patterns软件建立分类树鉴定模型,并进行盲法验证。结果:在相对分子质量(Mr)3 000～20 000范围内,捕获104个蛋白峰,其中90个蛋白峰差异有统计学意义(P<0.01),择优选出质荷比(M/Z)为10 061.7的蛋白峰建立甲型副伤寒沙门氏菌的分类树模型,甲型副伤寒沙门氏菌诊断的灵敏度和特异度通过盲法验证为100%。结论:采用SELDI-TOF MS技术建立的甲型副伤寒沙门氏菌的分类树诊断模型,可用于该菌的快速鉴定。
Objective: To establish the protein fingerprinting of Salmonella paratyphi A by surface enhanced laser desorption / ionization time of flight mass spectrometry. Methods: Thirty-six strains of Salmonella paratyphi A isolated from clinical isolates were collected and 96 control strains were collected simultaneously. The collected bacteria were verified by molecular biology using 16S rDNA sequencing. Bacterial proteins were detected by surface-enhanced laser desorption / ionization time-of-flight mass spectrometry. ProteinChip and Biomarker Wizard software were used to collect data automatically. BioMarker Patterns software was used to establish the classification tree identification model and validated by blind method. Results: 104 protein peaks were captured in the range of molecular weight (Mr) of 3000 to 20 000, of which 90 protein peaks were statistically significant (P <0.01), and the mass / charge ratio (M / Z) The classification tree model of Salmonella paratyphi A was established for the protein peak of 10 061.7. The sensitivity and specificity of the Salmonella paratyphi A diagnosis was 100% by blinded method. Conclusion: The classification tree diagnosis model of Salmonella paratyphi A established by SELDI-TOF MS technique can be used for the rapid identification of the bacteria.