肖迪, 秦天, 侯学新, 郜振国, 王冀涛, 张慧芳, 张炳华, 李明慧, 孙静轩, 王磊, 杨文涛, 李天一, 马合木提, 卢金星. 基于MALDI-TOF MS的新型冠状病毒感染快速诊断技术构建[J]. 疾病监测, 2021, 36(11): 1196-1202. DOI: 10.3784/jbjc.202110120543
引用本文: 肖迪, 秦天, 侯学新, 郜振国, 王冀涛, 张慧芳, 张炳华, 李明慧, 孙静轩, 王磊, 杨文涛, 李天一, 马合木提, 卢金星. 基于MALDI-TOF MS的新型冠状病毒感染快速诊断技术构建[J]. 疾病监测, 2021, 36(11): 1196-1202. DOI: 10.3784/jbjc.202110120543
Xiao Di, Qin Tian, Hou Xuexin, Gao Zhenguo, Wang Jitao, Zhang Huifang, Zhang Binghua, Li Minghui, Sun Jingxuan, Wang Lei, Yang Wentao, Li Tianyi, Muti-Mahe, Lu Jinxing. Establishment of a rapid diagnosis assay for SARS-CoV-2 infection based on MALDI-TOF MS[J]. Disease Surveillance, 2021, 36(11): 1196-1202. DOI: 10.3784/jbjc.202110120543
Citation: Xiao Di, Qin Tian, Hou Xuexin, Gao Zhenguo, Wang Jitao, Zhang Huifang, Zhang Binghua, Li Minghui, Sun Jingxuan, Wang Lei, Yang Wentao, Li Tianyi, Muti-Mahe, Lu Jinxing. Establishment of a rapid diagnosis assay for SARS-CoV-2 infection based on MALDI-TOF MS[J]. Disease Surveillance, 2021, 36(11): 1196-1202. DOI: 10.3784/jbjc.202110120543

基于MALDI-TOF MS的新型冠状病毒感染快速诊断技术构建

Establishment of a rapid diagnosis assay for SARS-CoV-2 infection based on MALDI-TOF MS

  • 摘要:
      目的  构建一种采用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)的新型冠状病毒(SARS-CoV-2)感染快速血清多肽指纹谱检测方法。
      方法  采用MALDI-TOF MS-ClinProTools系统,通过构建遗传算法(GA)分类数学模型进行SARS-CoV-2感染的诊断。 本研究共采用647份血清样本(228份来源于SARS-CoV-2感染阳性病例,419份来自感染阴性人员),其中160份(阳性、阴性样本各80份)用于构建分析模型,487份用于模型优化与验证,并采用非标定量差异蛋白质组技术对阳性和阴性血清样本进行蛋白水平的解析。
      结果  本研究构建的最优模型(GA模型)的交叉验证能力和识别能力分别是99.37%和100.00%,发现19个多肽标志物,对487份血清样本(148份阳性,339份阴性)鉴定的准确率为98.00%(阳性样本为100.00%,阴性样本为96.00%)。 差异蛋白质组学和生物信息学分析确定95个感染相关差异蛋白。 利用本研究构建的方法可在3 h内完成96份血清样本的检测,并且只需2滴指血即可满足检测要求。 血清在使用前灭活,无生物安全隐患,操作可在普通分子生物学实验室进行。
      结论  本研究为SARS-CoV-2感染筛查与诊断提供了一种具有巨大应用潜力的新型检测方法。

     

    Abstract:
      Objective  To establish a rapid, high-throughput detection assay of serum peptidome profiling for the diagnosis of SARS-CoV-2 infection based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS).
      Methods  MALDI-TOF MS-ClinProTools system was used to construct a genetic algorithm (GA)-based classification model to diagnose SARS-CoV-2 infection in this study. In total, 647 serum samples (228 positive ones, 419 negative ones) were used. Specifically, 160 serum samples (80 positive ones, 80 negative ones) were used for model construction and optimization, and 487 serum samples were used for model validation. Differential proteomics was used to analyze the differences in serum polypeptide between COVID-19 patients and non-COVID-19 patients.
      Results  For the GA screening model, the cross-validation values and the recognition capability values were 99.37% and 100.00%, respectively. Nineteen biomarker peptides were found. The identification accuracy of 487 samples (148 COVID-19 patients and 339 non-COVID-19 patients’ serum samples) was 98.00% (100.00% for positive samples, 96.00% for negative samples). Ninety-five significantly differentially regulated serum proteins were identified between the COVID-19 patients and non-COVID-19 patients by quantification analysis. The assay established in this study can detect 96 serum samples within 3 hours. Only 2 drops of finger blood can meet the detection requirements. The serum is inactivated before use, and there is no potential biosafety hazard. The operation can be carried out in the general molecular biology laboratory.
      Conclusion  This study established a new detection assay with great application potential for the screening and diagnosis of SARS-CoV-2 infection.

     

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