Objectives: To study new biomarkers for the early diagnosis of retinopathy of prematurity (ROP) by analyzing the differences in blood metabolites based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) and metabolomics.
Methods: Dried blood spots were collected from 21 infants with ROP (ROP group) and 21 infants without ROP (non-ROP group) who were hospitalized in the Sixth Affiliated Hospital of Sun Yat-sen University from January 2013 to December 2016. LC-MS/MS was used to measure the metabolites, and orthogonal partial least squares-discriminant analysis was used to search for differentially expressed metabolites and biomarkers.
Results: There was a significant difference in blood metabolic profiles between the ROP and non-ROP groups. The pattern recognition analysis, Score-plot, and weight analysis obtained 10 amino acids with a relatively large difference. Further statistical analysis showed that the ROP group had significant increases in blood levels of glutamic acid, leucine, aspartic acid, ornithine, and glycine compared with the non-ROP group (P<0.05). The receiver operating characteristic curve analysis showed that glutamic acid and ornithine had the highest value in diagnosing ROP.
Conclusions: Blood metabolites in preterm infants with ROP are different from those without ROP. Glutamic acid and ornithine are the metabolic markers for diagnosing ROP. LC-MS/MS combined with metabolomics analysis has a potential application value in the early identification and diagnosis of ROP.
目的: 应用液相色谱-串联质谱技术（liquid chromatography tandem mass spectrometry，LC-MS/MS）和代谢组学方法探讨早产儿视网膜病（retinopathy of prematurity，ROP）患儿出生血代谢产物差异，寻找ROP早期诊断的新的生物标记物。方法: 收集2013年1月—2016年12月中山大学附属第六医院住院的21例ROP患儿（ROP组）及同期21例非ROP患儿（非ROP组）干血片标本，利用LC-MS/MS进行代谢产物测定，运用正交偏最小二乘判别分析法（orthogonal partial least squares discriminant analysis，OPLS-DA）寻找差异物质和生物标记物。结果: ROP组和非ROP组患儿血代谢谱有明显差异，经模式识别分析、代谢物得分图（Score-plot，S-plot）、权重分析初步得出10个差异较大的氨基酸。进一步统计分析发现ROP组患儿血谷氨酸、亮氨酸、天冬氨酸、鸟氨酸和甘氨酸水平明显高于非ROP组，差异有统计学意义（P<0.05）。受试者工作特征曲线分析显示，谷氨酸及鸟氨酸对ROP诊断价值最高。结论: ROP患儿与非ROP患儿比较血代谢产物具有明显差异，谷氨酸及鸟氨酸是诊断ROP的代谢标志物。LC-MS/MS结合代谢组学分析方法在ROP早期识别与诊断中具有潜在的应用价值。.
Keywords: Biomarker; Liquid chromatography; Metabolomics; Preterm infant; Retinopathy of prematurity; Tandem mass spectrometry.