甲型副伤寒沙门氏菌蛋白指纹图谱的建立

Establishment of protein fingerprint database of Salmonella paratyphi A using SELDI-TOF-MS

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。

AIM： To establish a protein fingerprint database of Salmonella paratyphi A by surface enhanced laser desorption/ionization time-of-flight mass spectrometry （SELDI-TOF-MS）. METHODS： Thirty-six clinical bacterial isolates and 96 control bacteria isolates were collected and identified using 16S rDNA sequencing. Bacterial proteins were detected by SELDI-TOF-MS, and all protein finger- prints were analyzed by ProteinChip and Biomarker Wizard software. The analysis results were used to set up a classifi- cation tree model by means of BioMarker Patterns software. At the same time, the data were tested by a blinded validation. RESULTS： In the range of Mr 3 000 -20 000, we obtained 104 protein peaks, of which 90 were of statistical significance （ P 〈 0.01 ）. A protein peak with mass-to-charge ratio （M/Z） l0 061.7 was chosen to establish the classification tree mod- el of Salmonella paratyphi A, and the sensitivity and speci- ficity of Salmonella paratyphi A diagnosis was 100% as shown by the blinded validation. CONCLUSION： The classification tree model of Salmonella paratyphi A can be not only established using SELDI-TOF-MS technology, but also used for the rapid identification of Salmonella paratyphi A.