目的 对比分析荧光分析法(fluorescence assay,FA)和液相色谱串联质谱法(LC-MS/MS)用于新生儿高苯丙氨酸血症(hyperphenylalaninemia,HPA)筛查中的差异,为筛查HPA提供参考。方法 收集2016年10月至2018年9月新生儿足底滤纸干血片标本12 529份。分别采用FA和LC-MS/MS检测滤纸干血片的血苯丙氨酸(phenylalanine,Phe)浓度,LC-MS/MS检测酪氨酸(Tyr)浓度。对筛查的HPA阳性结果,通过检测患儿新鲜尿液、末梢血制成滤纸干尿片、干血片,测定尿蝶呤谱、红细胞二氢蝶啶还原酶活性,并进行基因二代测序予以明确HPA诊断。结果 两种筛查方法共发现HPA初筛阳性43例,其中FA检测方法发现43例,LC-MS/MS检测结果为4例,后者的4例初筛阳性者已被FA检测发现。两种筛查方法阳性检出率差异有统计学意义(χ2=32.423,P＜0.05);假阳性率分别为32.72/万(41/12 529)和1.59/万(2/12 529),43例初筛阳性者共确诊HPA 2例。两种检测方法发现的41例疑似HPA患儿经随访1年,均予以排除。结论 两种检测方法同样适应用于新生儿HPA的筛查, LC-MS/MS检测方法可降低样本的复检率,节约社会成本。
Abstract
Objective The difference between fluorescence assay (FA) and liquid chromatography-tandem mass spectrometry (LC-MS / MS) in neonatal hyperphenylalaninemia (HPA) screening was analyzed to provide reference for HPA screening. Methods From October 2016 to September 2018,a total of 12529 dried blood specimens of neonatal plantar filter paper were collected.FA and LC-MS / MS were used to detect the phenylalanine (Phe) concentration in dried blood specimens of filter paper,and LC-MS / MS was used to detect the tyrosine (Tyr) concentration.By detecting the dry filter papers made of fresh urine and peripheral blood of the children,the urinary pterin profile and the activity of dihydropteridine reductase of red blood cells were measured,and the gene was sequenced for the second generation,so as to make clear the diagnosis of HPA for the positive results of HPA screening. Results 43 cases of HPA were found positive by the two screening methods, of which 43 cases were detected by FA, 4 cases by LC-MS / MS, and 4 cases by FA. The positive rate of the two screening methods was statistically significant (χ2=32.423, P < 0.05); the false positive rate was 32.72/10 000 (41/12 529) and 1.59/10000 (2 / 12 529) respectively. 43 cases with positive primary screening were diagnosed with Hpa2. 41 cases of suspected HPA detected by the two methods were excluded after 1 year follow-up. Conclusion Both methods were suitable for HPA screening in neonates,LC-MS / MS detection method could reduce the sample retest rate and save the social cost.
关键词
高苯丙氨酸血症 /
荧光分析法 /
液相色谱串联质谱法 /
新生儿筛查 /
遗传代谢性疾病
Key words
Hyperphenylalaninemia /
Fluorescence analysis /
Liquid chromatography tandem mass spectrometry /
Neonatal screening /
Inherited metabolic diseases
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参考文献
[1] 顾学范,主编.临床遗传代谢病[M].北京:人民卫生出版社,2015,1-2.
[2] 中华医学会儿科学分会内分泌遗传代谢学组,中华预防医学会出生缺陷预防与控制专业委员会新生儿筛查学组.高苯丙氨酸血症的诊治共识[J].中华儿科杂志,2014,52(6):420-425.
[3] Sørensen LK,Johannsen M.Sensitive determination of monoamine neurotransmitters,their main metabolites and precursor amino acids in different mouse brain components by liquid chromatography-electrospray tandem mass spectrometry after selective sample clean-up[J].Biomed Chromatogr,2019,33(4):e4479.
[4] Didycz B,Bik-Multanowski M.Dynamics of hyperphenylalaninemia and intellectual outcome in teenagers with phenylketonuria[J].Acta Biochim Pol,2017,64(3):527-531.
[5] Rajabi F,Levy HL.Hyperphenylalaninemia and the genomic revolution[J].Mol Genet Metab,2015,114(3):380-381.
[6] 沈玉燕,黎剑,肖刚.应用串联质谱技术进行新生儿高苯丙氨酸血症79205例筛查结果分析[J].中国优生与遗传杂志,2018,26(8):65-66.
[7] HACE D H,SHERWIN J E,HILLMAN S L,et a1.Use of phenylalanine-to-tyrosine ratio determined by tandem mass spectrometry to improve newborn screening for phenylketonuria of early discharge specimens collected in the first 24 hours[J].Clin Chem,1998,44(12):2 405-2 409.
[8] 陈曦,付晖.串联质谱方法在苯丙酮尿症的实验诊断应用[J].兵团医学,2012,34(4): 17-18.
[9] 赵海燕,韩宁宁,刘志娟,等.比较四种检测方法在高苯丙氨酸血症诊断中的应用[J].中国优生与遗传杂志,2017,25(9):25-26+31.
[10] 王兴,徐瑞峰,刘芙蓉,等.串联质谱法新生儿高苯丙氨酸血症筛查的切值探讨[J].中国优生与遗传杂志,2017,25(7):84-85,22.
[11] 刘伟,马志军,简永健,等.串联质谱法和荧光分析法筛查苯丙酮尿症的对比分析[J].中国妇幼保健,2018,33(24):6020-6022.
[12] 刘鸿丽,李风侠.2010~2016年陕西省新生儿疾病筛查治疗及随访结果分析[J].中国儿童保健杂志,2018,26(11):1221-1223.
[13] 田国力,王燕敏,许洪平,等.串联质谱法和荧光分析法检测滤纸干血片苯丙氨酸的比较[J].检验医学,2016,31(9):814-819.
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