串联质谱与NGS在遗传代谢病诊断性能评估对比及评估其影响因素

Comparison of tandem mass spectrometry and NGS in the diagnosis of genetic metabolic diseases and evaluation of its influencing factors

目的比较串联质谱检测与二代测序技术(NGS)诊断疑似遗传代谢病(IMD)的新生儿的性能,并分析影响其诊断的可能因素。方法纳入2019年8月至2022年6月承德市妇幼保健院临床表现疑似IMD的120例新生儿,均接受参加串联质谱检测和NGS诊断。统计分析不同诊断方法的IMD分类情况,比较两种方法的诊断效能。比较不同胎龄(早产、足月)、出生体质量(≤2500 g、>2500 g)、采血时间(3~7 d、>7 d)新生儿氨基酸浓度。结果120例疑似IMD新生儿中,经串联质谱诊断判断阳性病例有27例(22.50%),其中氨基酸代谢病13例、脂肪酸代谢病6例、有机酸代谢病8例。120例疑似IMD新生儿中,经NGS诊断判断阳性病例有25例(20.83%),其中氨基酸代谢病12例、脂肪酸代谢病7例、有机酸代谢病6例。以NGS测序结果为金标准,串联质谱诊断准确率为93.33%(112/120),敏感度、特异度分别为88.00%(22/25)、94.74%(90/95)。不同胎龄(早产、足月)的丙氨酸(Ala)、甘氨酸(Gly)、甲硫氨酸(Met)、鸟氨酸(Orn)、脯氨酸(Pro)、亮氨酸(Leu)、酪氨酸(Tyr)、缬氨酸(Val)浓度差异有统计学意义,不同出生体质量(≤2500 g、>2500 g)的Ala、Orn、Pro、Tyr、Val浓度差异有统计学意义,不同采血时间(3~7 d、>7 d)Ala、Arg、Cit、Gly、Met、Orn、Leu、Val浓度差异有统计学意义(P<0.05)。结论串联质谱技术能有效诊断多种遗传代谢病,但诊断性能容易受到新生儿胎龄、出生体质量、采血时间等因素的影响,串联质谱联合NGS可提高新生儿IMD的诊断效能。

Objective To compare the diagnostic performance of tandem mass spectrometry and next generation sequencing(NGS)in diagnostic newborn infants with inherited metabolic dieases(IMD),and analyze the possible factors affecting tandem mass spectrometry screening.Methods From August 2019 to June 2022,120 neonates with clinical manifestations suspected of IMD in the Chengde Maternal and Child Health Care Hospital were enrolled,and they all received tandem mass spectrometry and NGS screening.The IMD classification of different diagnostic methods was statistically analyzed,and the diagnostic efficacy of the two methods was compared.The amino acid concentrations of neonates at different gestational ages(premature and full-term),birth weight(≤2500 g,>2500 g),and blood collection time(3–7 d,>7 d)were compared.Results Among 120 suspected IMD neonates,27(22.50%)were positive by tandem mass spectrometry screening,including 13amino acid metabolic diseases,6 fatty acid metabolic diseases,and 8 organic acid metabolic diseases.Among 120 suspected IMD neonates,25(20.83%)were diagnosed as positive by NGS screening,including 12 amino acid metabolic diseases,7 fatty acid metabolic diseases and 6 organic acid metabolic diseases.Using NGS sequencing results as the gold standard,the accuracy of tandem mass spectrometry diagnostic was 93.33%(112/120),the sensitivity and specificity were 88.00%(22/25)and 94.74%(90/95),respectively.The concentrations of Ala,Gly,Met,Orn,Pro,Leu,Tyr,Val in different gestational ages(preterm and full-term)were statistically significant,the concentrations of Ala,Orn,Pro,Tyr,Val in different birth weight(≤2500 g,>2500 g)were statistically significant.The concentrations of Ala,Arg,Cit,Gly,Met,Orn,Leu and Val at different blood collection times(3–7 d,>7 d)were statistically significant(P<0.05).Conclusion Tandem mass spectrometry can effectively screen a variety of genetic metabolic diseases,but the diagnostic performance is easily affected by neonatal gestational age,birth weight,blood collection time and other factors.Tandem mass spectrometry combined with NGS can improve the diagnostic efficiency of neonatal IMD.