文献类型：期刊文献

中文题名：超高效液相色谱-串联质谱法测定人血清中结核病生物标志物CFP-10

英文题名：Determination of tuberculosis biomarker CFP-10 in serum by ultra-performance liquid chromatography-tandem mass spectrometry

作者：方媚[1];许文芳[2];王娇娇[1];吴筱丹[1];金法祥[2];李士敏[1,3]

机构：[1]浙江大学农生环测试中心,杭州310058;[2]绍兴文理学院附属医院检验科,绍兴312000;[3]泽达精准(杭州)生物科技有限公司,杭州310051

年份：2025

卷号：44

期号：4

起止页码：539

中文期刊名：分析试验室

外文期刊名：Chinese Journal of Analysis Laboratory

收录：北大核心2023、、EI(收录号：20252318539350)、北大核心

基金：浙江省教育厅科研项目(Y202043226)资助。

语种：中文

中文关键词：培养滤液蛋白10;超高效液相色谱-串联质谱法;结核病;血清

外文关键词：culture filtrate protein 10;ultra performance liquid chromatography-tandem mass spectrometry;tuberculosis;serum

中文摘要：建立了超高效液相色谱-串联质谱(UPLC-MS/MS)测定人血清中结核病(TB)生物标志物培养滤液蛋白10(CFP-10)的方法。血清样品经蛋白沉淀、胰蛋白酶消化后,采用固相萃取(SPE)富集纯化CFP-10酶解产生的特异性肽段,经Luna Omega PS-C18色谱柱分离后,采用电喷雾电离源正离子模式(ESI+),多反应监测(MRM)模式检测。结果表明:方法在3~100 ng/mL范围内线性良好,线性相关系数r2=0.9906,方法检出限和定量限分别为1.0和3.0 ng/mL,加标回收率为89.8%~108.1%,日内和日间的相对标准偏差均低于10%。采用该方法对61个实际血清样品进行测定,结果显示结核病例组的血清CFP-10水平较非结核病对照组显著升高。该方法可实现高通量处理,适用于临床血清样本中CFP-10的检测。

外文摘要：An ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method was developed to determine the tuberculosis(TB)biomarker CFP-10 in human serum.After protein precipitation and trypsin digestion,solid-phase extraction(SPE)was used to enrich the specific peptide produced during enzymatic digestion of culture filtrate protein 10(CFP-10).The separation of the target analyte was performed on a Luna Omega PS-C18 column.The specific CFP-10 peptide was quantified under positive electrospray ion electrospray ionization(ESI+)and multiple reaction monitoring(MRM)mode.The results showed that the calibration curve was linear in the range of 3.0-100 ng/mL for CFP-10,with the correlation coefficient of r2=0.9906.The limit of detection(LOD)and limit of quantification(LOQ)for CFP-10 were 1.0 and 3.0 ng/mL,respectively.The spiked recoveries ranged from 89.8%to 108.1%,with the intra-day and inter-day relative standard deviations(RSDs)both lower than 10%.Serum CFP-10 levels in TB and non-TB control cases were compared,and the results showed that serum CFP-10 was significantly lower in TB control than in non-TB cases.The proposed method is suitable for the rapid determination of CFP-10 in clinical human serum samples.