【摘要】 目的 探讨串联质谱法(MS/MS)筛查甲基丙二酸血症(MMA)的基础上联合高效液相色谱-串联质谱法(LC-MS/MS)、气相色谱质谱法(GC-MS)诊断MMA的价值。方法 2015年5月—2023年3月上海市儿童医院诊治MMA患儿39例为观察组，同期健康体检儿童200例为对照组。2组采用MS/MS法检测血C3、C2、C3/C2、MET;采用LC-MS/MS法检测血Mma、MET、MCA和Hcy;采用尿素酶预处理GC-MS法检测尿3-HP、Mma、MCA;绘制ROC曲线，评估MS/MS、LC-MS/MS、GC-MS联合诊断MMA的效能。结果 (1)观察组MS/MS法检测的C2[13.04(7.63,24.58)μmol/L]、C3[6.38(4.17,11.85)μmol/L]、C3/C2[0.56(0.34,0.90)]均高于对照组[9.82(7.10,13.45)μmol/L、1.04(0.70,1.50)μmol/L、0.10(0.08,0.13)](U=2 986.0、102.0、378.0,P均<0.05),MET[9.58(7.94,14.90)μmol/L]低于对照组[22.59(17.34,26.77)μmol/L](U=1 533.0,P<0.001)。(2)观察组LC-MS/MS法检测的Mma[31.23(14.80,55.42)μmol/L]、MCA[1.32(0.80,2.40)μmol/L]和Hcy[66.31(11.60,97.11)μmol/L]均高于对照组[0.60(0.40,0.81)、0.20(0.10,0.30)、4.60(3.80,5.70)μmol/L](U=75.5、483.5、814.0,P均<0.001),MET[7.10(3.70,19.20)μmol/L]低于对照组[29.10(22.60,36.35)μmol/L](U=706.0,P<0.001)。(3)观察组GC-MS法检测的3-HP[4.50(1.62,14.14)mmol/mmol Cr]、Mma[93.80(38.81,220.70)mmol/mmol Cr]、MCA[2.30(1.13,5.22)mmol/mmol Cr]均高于对照组[0.51(0.29,0.87)、0、0.58(0.37,0.94)mmol/mmol Cr](U=827.0、139.5、306.5,P均<0.05)。(4)ROC曲线分析结果显示，C3、C3/C2和MET分别以2.616 5μmol/L、0.222 0、14.925 0μmol/L为最佳截断值，联合筛查MMA的AUC为0.992(95%CI:0.983～1.000,P<0.001),灵敏度为97.4%,特异度为96.0%;MCA、Hcy、Mma、MET分别以0.74、10.55、3.28、12.98μmol/L为最佳截断值，联合筛查MMA的AUC为1.000(95%CI:0.999～1.000,P<0.001),灵敏度为100.0%,特异度为99.5%;3-HP、Mma、MCA分别以1.535、6.750、0.165 mmol/mmol Cr为最佳截断值，联合筛查MMA的AUC为0.982(95%CI:0.948～1.000,P<0.001),灵敏度为97.4%,特异度为100.0%。3种方法联合诊断MMA的AUC为1.000(95%CI:0.948～1.000,P<0.001),灵敏度为100.0%,特异度为100.0%。结论 MMA患者C3、C3/C2、Mma、MCA、3-HP明显增高。MS/MS、LC-MS/MS、GC-MS联合诊断MMA的准确率高。更多还原

【Abstract】 Objective To explore the value of combination of tandem mass spectrometry(MS/MS),high-performance liquid chromatography MS/MS(LC-MS/MS) and gas chromatogramphy MS(GC-MS) to the diagnosis of methylmalonic acidemia(MMA).Methods Thirty-nine children with MMA diagnosed in Shanghai Children’s Hospital from May 2015to March 2023 were included as observation group,and 200 children receiving physical examination in the same period were as the controls(control group).MS/MS was used to detect the levels of C3,C2,C3/C2 and MET in two groups.LC-MS/MS was used to detect the levels of Mma,MET,MCA and Hey.Urease pretreatment GC-MS method was used to detect the levels of 3-HP,Mma and MCA from urine specimens.ROC curves were plotted to observe the value of combination of MS/MS,LC-MS/MS and GC-MS to the diagnosis of MMA.Results(1) C2,C3 and C3/C2 were higher in observation group [13.04(7.63,24.58) μmol/L,6.38(4.17,11.85) μmol/L,056(0.34,0.90)] than those in control group [9.82(7.10,13.45) μmol/L,1.04(0.70,1.50) μmol/L,0.10(0.08,0.13)](U=2 986.0,102.0,378,0;all P values <0.05),and MET was lower in observation group [9.58(7.94,14.90) μmol/L] than that in control group [22.59(17.34,26.77) μmol/L](U=1 533.0,P<0.001).(2) LC-MS/MS showed that the levels of Mma,MCA and Hcy were higher in observation group [31.23(14.80,55.42),1.32(0.80,2.40),66.31(11.60,97.11) μmol/L]than those in control group [0.60(0.40,0.81),0.20(0.10,0.30),4.60(3.80,5.70) μmol/L](U=75.5,483.5,814.0;all P values <0.001),and MET was lower in observation group [7.10(3.70,19.20) μmol/L] than that in control group [29.10(22.60,36.35) μmol/L](U=706.0,P<0.001).(3) GC-MS showed that the levels of 3-HP,Mma and MCA were higher in observation group [4.50(1.62,14.14),93.80(38.81,220.70),2.30(1.13,5.22) mmol/mmol Cr] than those in control group [0.51(0.29,0.87),0,0.58(0.37,0.94) mmol/mmol Cr](U=827.0,139,5,306.5;all P values <0.05).(4) ROC curve analysis showed that with 2.616 5 μmol/L,0.222 0and 14.925 0 μmol/L as the optimal cut-off values of C3,C3/C2 and MET,the AUC of combination of them three for diagnosing MMA was 0.992(95%CI:0.983-1.000,P<0.001),with a sensitivity of 97.4%,and specificity of 96.0%;with 0.74,10.55,3.28 and 12.98 μmol/L as the optimal cut-off values of MCA,Hcy,Mma and MET,the AUC of the combination of them four was 1.000(95 %CI:0.999-1.000, P<0.001),with a sensitivity of 100.0% and specificity of99.5%;with 1.535,6.750 and 0.165 mmol/mmol Cr as the optimal cut-off values of 3-HP,Mma and MCA,the AUC of the combination of them three was 0.982(95% CI:0.948-1.000,P<0.001),with a sensitivity of 97.4% and a specificity of 100.0%.The AUC of MS/MS,LC-MS/MS and GC-MS was 1.000(95%CI:0.948-1.000,P<0.001),with a sensitivity of 100.0% and a specificity of 100.0%,Conclusion The C3,C3/C2,Mma,MCA and 3-HP increase in MMA children,and the combination of MS/MS,LC-MS/MS and GC-MS has a high accuracy rate for diagnosing MMA.更多还原