DP-TOF 质谱检测的准确性及临床应用。

Detection accuracy and clinical applications of DP-TOF mass spectrometry.

机构信息

State Key Laboratory of Cardiovascular Disease, Beijing Key Laboratory for Molecular Diagnostics of Cardiovascular Diseases, Center of Laboratory Medicine, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.

Center of Laboratory Medicine, Fuwai Yunnan Cardiovascular Hospital, Kunming, Yunnan, China.

出版信息

J Int Med Res. 2024 May;52(5):3000605241255568. doi: 10.1177/03000605241255568.

DOI:10.1177/03000605241255568

PMID:38819085

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11143829/

Abstract

OBJECTIVE

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is currently used in clinical microbiology laboratories. This study aimed to determine whether dual-polarity time-of-flight mass spectrometry (DP-TOF MS) could be applied to clinical nucleotide detection.

METHODS

This prospective study included 40 healthy individuals and 110 patients diagnosed with cardiovascular diseases. We used DP-TOF MS and Sanger sequencing to evaluate 17 loci across 11 genes associated with cardiovascular drug responses. In addition, we used DP-TOF MS to test 998 retrospectively collected clinical DNA samples with known results.

RESULTS

A, T, and G nucleotide detection by DP-TOF MS and Sanger sequencing revealed 100% concordance, whereas the C nucleotide concordance was 99.86%. Genotyping based on the results of the two methods showed 99.96% concordance. Regarding clinical applications, DP-TOF MS yielded a 99.91% concordance rate for known loci. The minimum detection limit for DNA was 0.4 ng; the inter-assay and intra-assay precision rates were both 100%. Anti-interference analysis showed that aerosol contamination greater than 10 copies/µL in the laboratory environment could influence the results of DP-TOF MS.

CONCLUSIONS

The DP-TOF MS platform displayed good detection performance, as demonstrated by its 99.96% concordance rate with Sanger sequencing. Thus, it may be applied to clinical nucleotide detection.

摘要

目的

基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）目前用于临床微生物学实验室。本研究旨在确定双极性飞行时间质谱（DP-TOF MS）是否可应用于临床核苷酸检测。

方法

本前瞻性研究纳入了 40 名健康个体和 110 名诊断为心血管疾病的患者。我们使用 DP-TOF MS 和 Sanger 测序来评估与心血管药物反应相关的 11 个基因中的 17 个位点。此外，我们使用 DP-TOF MS 测试了 998 份回顾性收集的已知结果的临床 DNA 样本。

结果

DP-TOF MS 和 Sanger 测序的 A、T 和 G 核苷酸检测的一致性为 100%，而 C 核苷酸的一致性为 99.86%。基于两种方法的结果进行基因分型显示，一致性为 99.96%。在临床应用方面，DP-TOF MS 对已知位点的符合率为 99.91%。DNA 的最小检测限为 0.4ng；批内和批间精密度率均为 100%。抗干扰分析表明，实验室环境中气溶胶污染大于 10 拷贝/µL 可能会影响 DP-TOF MS 的结果。