液相色谱串联质谱技术在新生儿先天性肾上腺皮质增生症筛查的运用  

Application of liquid chromatography tandem mass spectrometry in newborn screening for congenital adrenal hyperplasia

作　　者：苏跃青[1] 朱文斌[1] 周进福[1] 陈涵强[1] SU Yueqing;ZHU Wenbin;ZHOU Jinfu;CHEN Hanqiang(Fujian Maternity and Children Health Hospital,Affiliated Hospital of Fujian Medical University,Fuzhou,Fujian 350001,China)

机构地区：[1]福建医科大学附属医院福建省妇幼保健院,福州350001

出　　处：《福建医药杂志》2020年第1期13-15,共3页Fujian Medical Journal

基　　金：福建省自然科学基金资助项目(2016J01489)。

摘　　要：目的采用液相色谱串联质谱(LC-MS/MS)技术,对传统新生儿先天性肾上腺皮质增生症(CAH)筛查方法进行改进。方法对73例时间分辨荧光法(TRFIA)初筛异常的疑似CAH患儿,先用基因分析技术为诊断标准分为CYP21A2基因突变患者、携带者和非CAH21新生儿,再用LC-MS/MS测定17-羟孕酮(17-OHP)和21-脱氧皮质醇(21-DF)浓度,最后比较LC-MS/MS技术与TRFIA法的差异。结果73例研究对象中,CAH21患者6例,携带者9例,非CAH21新生儿58例。LC-MS/MS法的17-OHP浓度1.65~218.63ng/mL,21-DF浓度0.112~23.300ng/mL;其中,CAH21患儿的17-OHP浓度显著高于携带者或非CAH21新生儿(均P<0.001);21-DF浓度:CAH21患儿>携带者>非CAH21新生儿(分别P<0.001和P<0.05)。Deming回归和Bland-Altman偏差分析:LC-MS/MS技术与TRFIA法检测结果的关联性良好,但后者明显高于前者。根据LC-MS/MS技术检测正常人群上述两指标的参考范围,前者假阳性27例,为TRFIA法40.30%;后者假阳性13例,为TRFIA法19.40%。结论LC-MS/MS技术应用对初筛阳性的疑似CAH患儿进行原干血片测定17-OHP和21-DF浓度,可降低TRFIA过高的假阳性率。Objective To improve newborn screening method for congenital adrenal hyperplasia(CAH21)using liquid chromatography-tandem mass spectrometry(LC-MS/MS)method.Methods In total,73CAH suspects using classical timeresolved fluoroimmunoassay(TRFIA)were enrolled and collected their original dry blood samples.CYP21A2mutations among all participants were assessed as confirmatory tests for CAH21patients.LC-MS/MS method was used to further measure the 17-hydroxyprogesterone(17-OHP)and 21-deoxycortisol(21-DF).Results Gene analysis confirmed that there were 6CAH21patients,9carriers and 58newborns of none CYP21A2mutation in73CAH suspects.The concentrations of 17-OHP and 21-DF in all participants using LC-MS/MS were range from 1.65ng/mL to 218.63ng/mL and 0.112ng/mL to 23.300ng/mL,respectively.Comparing with carriers or newborns of none CYP21A2mutation,the 17-OHP in CAH patients increased significantly(both P<0.001).Whereas,concentration of 21-DF in CAH patients>mutation carriers>CAH21negative newborns(P<0.001and P<0.05,respectively).Deming regression analysis method and Bland-Altman plot showed that there was a good correlation in 17-OHP concentrations between using LC-MS/MS and using TRFIA.According the reference range of 17-OHP and 21-DF by LC-MS/MS in normal newborns,40.30%and 19.40%of false-positive results were remained,compared with that by TRFIA,respectively.Conclusion Additional test of 17-OHP and 21-DF in original dried blood spots of newborns who have a positive initial screening using LC-MS/MS method are effective to reduce the ratio of false positive.

关 键 词：LC-MS/MS技术 先天性肾上腺皮质增生症 17-羟孕酮 21-脱氧皮质醇 

分 类 号：R722.1[医药卫生—儿科]