递送及储存条件对新生儿干血斑标本中氨基酸和肉碱浓度的影响

doi:10.3785/j.issn.1008-9292.2020.10.03

浙江大学学报（医学版）

2020, Vol. 49

Issue (5): 565-573 DOI: 10.3785/j.issn.1008-9292.2020.10.03

专题报道

递送及储存条件对新生儿干血斑标本中氨基酸和肉碱浓度的影响

胡凌微¹(

),胡真真¹,杨建滨¹,张玉²,施叶珍²,祝莎莎³,杨茹莱¹,黄新文^1,*(

)

1. 浙江大学医学院附属儿童医院遗传与代谢科国家儿童健康与疾病临床医学研究中心国家儿童区域医疗中心, 浙江杭州 310052
2. 浙江博圣生物技术股份有限公司, 浙江杭州 310012
3. 台州市妇女儿童医院儿科, 浙江台州 318000

Effects of delivery and storage conditions on concentrations of amino acids and carnitines in neonatal dried blood spots

HU Lingwei¹(

),HU Zhenzhen¹,YANG Jianbin¹,ZHANG Yu²,SHI Yezhen²,ZHU Shasha³,YANG Rulai¹,HUANG Xinwen^1,*(

)

1. Department of Genetics and Metabolism, Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, National Regional Medical Center for Children, Hangzhou 310052, China
2. Zhejiang Biosan Biochemical Technologies Co., Ltd., Hangzhou 310012, China
3. Department of Pediatrics, Taizhou Maternal and Child Health Hospital, Taizhou 318000, Zhejiang Province, China

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摘要：

目的: 探讨不同递送和储存条件对新生儿干血斑标本中氨基酸和肉碱浓度的影响，为提高串联质谱筛查新生儿遗传代谢病的准确性及可靠性提供依据。方法: 根据不同递送和储存条件将1 254 616名在浙江省新生儿疾病筛查中心进行遗传代谢病筛查新生儿的干血斑标本分为3组：室温递送储存（室温组，n=338 467）、冷链递送+低温储存（冷链+低温组，n=480 021）、冷链递送+低温低湿储存（冷链+低温低湿组，n=436 128）。应用串联质谱技术检测干血斑标本中氨基酸和肉碱浓度，SPSS 24.0软件进行数据分析，分析递送和储存过程中温度和湿度对氨基酸和肉碱浓度稳定性的影响。结果: 三组样本氨基酸和肉碱浓度均呈偏态分布，氨基酸和肉碱浓度组间差异均有统计学意义（均P < 0.01）。酪氨酸浓度中位数冷链+低温组与冷链+低温低湿组相近，室温组较冷链+低温组和冷链+低温低湿组分别降低18%和16%；室温组的蛋氨酸浓度中位数与冷链+低温组相近，分别较冷链+低温低湿组降低15%和11%；3组精氨酸浓度中位数均有差异，室温组和冷链+低温组分别较冷链+低温低湿组降低12%和25%。室温组的游离肉碱浓度中位数与冷链+低温组相近，较冷链+低温低湿组增高12%。冷链+低温组的乙酰肉碱、丙酰肉碱、丙二酰肉碱+3-羟基-丁酰肉碱、戊二酰肉碱+3-羟基-己酰肉碱、十六碳酰肉碱等浓度中位数与冷链+低温低湿组相近，室温组较冷链+低温组和冷链+低温低湿组降低21%~64%。其他氨基酸和酰基肉碱浓度组间差异较小。除瓜氨酸、甲基丙二酰肉碱+3-羟基-异戊酰肉碱、戊酰基肉碱外，室温组其他氨基酸和肉碱浓度月中位数变异系数均高于冷链+低温组和冷链+低温低湿组。结论: 使用冷链递送和低温低湿储存可以有效延缓干血斑标本中部分氨基酸和肉碱的降解速度，提高串联质谱检测结果的准确性和可靠性，从而保证新生儿遗传代谢病筛查质量。

关键词： 干血斑检测; 冷链运输; 储存; 温度; 湿度; 氨基酸类; 肉碱; 遗传代谢病筛查

Abstract:

Objective: To explore effects of different delivery and storage conditions on concentrations of amino acids and carnitines in neonatal dried blood spots (DBS), so as to provide evidence for improving accurate and reliable detection by tandem mass spectrometry. Methods: A total of 1 254 616 newborn DBS samples in Newborn Screening Center of Zhejiang Province were delivered and stored at room temperature (group A, n=338 467), delivered by cold-chain logistics system and stored at low temperature (group B, n=480 021), or delivered by cold-chain logistics system and stored at low temperature and low humidity (group C, n= 436 128), respectively. The concentrations of amino acids and carnitines in DBS were detected by tandem mass spectrometry. Data analysis was performed by SPSS 24.0 to explore the influence of temperature and humidity on the concentrations of amino acids and carnitines. Results: The concentrations of amino acids and carnitines in the three groups were skewed, and the differences in amino acid and carnitine concentrations among groups were statistically significant (all P < 0.01). The median concentration of tyrosine was lower in group A than those in group B and group C by 18%and 16%respectively, while there was no significant difference between the last two groups. The median concentrations of methionine were lower in group A and group B than that in group C by 15%and 11%, respectively. The median concentrations of arginine were lower in group A and group B than that in group C by 12%and 25%, respectively. The median concentration of free carnitine (C0) was higher in group A than that in group C by 12%, while there was no significant difference between group A and group B. The median concentrations of acetylcarnitine (C2), propionyl carnitine (C3), C3DC+C4OH, C5DC+C6OH and hexadecanoyl carnitine (C16) were lower in group A than those in group B and group C by 21%-64%. The concentrations of other amino acids and acylcarnitines differed little among three groups. The monthly median coefficients of variation of other amino acids and carnitines in group A were higher than those in group B and group C except for citrulline, C4DC+C5OH and isovalerylcarnitine (C5). Conclusion: Cold-chain logistics system and storage in low temperature and low humidity can effectively reduce degradation of some amino acids and carnitines in DBS, improve the accuracy and reliability of detection, and thus ensures the quality of screening for neonatal metabolic diseases.

Key words: Dried blood spot testing Cold-chain transportation Storage Temperature Humidity Amino acids Carnitine Genetic and metabolic diseases screening

收稿日期: 2020-05-12 出版日期: 2020-11-19

R446.1

基金资助: 国家重点研发计划(2018YFC1002204);浙江省公益技术研究计划(LGC19B050013)

通讯作者: 黄新文 E-mail: hulw@zju.edu.cn;6305022@zju.edu.cn

作者简介: 胡凌微(1991-), 女, 学士, 技师, 主要从事儿童遗传代谢病筛查工作; E-mail:hulw@zju.edu.cn; https://orcid.org/0000-0001-8442-0579

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引用本文:

胡凌微,胡真真,杨建滨,张玉,施叶珍,祝莎莎,杨茹莱,黄新文. 递送及储存条件对新生儿干血斑标本中氨基酸和肉碱浓度的影响[J]. 浙江大学学报（医学版）, 2020, 49(5): 565-573.

HU Lingwei,HU Zhenzhen,YANG Jianbin,ZHANG Yu,SHI Yezhen,ZHU Shasha,YANG Rulai,HUANG Xinwen. Effects of delivery and storage conditions on concentrations of amino acids and carnitines in neonatal dried blood spots. J Zhejiang Univ (Med Sci), 2020, 49(5): 565-573.

链接本文:

http://www.zjujournals.com/med/CN/10.3785/j.issn.1008-9292.2020.10.03 或 http://www.zjujournals.com/med/CN/Y2020/V49/I5/565

表 1 三组新生儿干血斑标本递送和储存条件

表 2 不同递送和储存条件下1 254 616名新生儿干血斑标本中各种氨基酸浓度

图 1 不同递送和储存条件下1 254 616名新生儿干血斑标本中各种氨基酸浓度月中位数变化趋势图

组别	游离肉碱		乙酰肉碱		丙酰肉碱		丁酰肉碱		丙二酰肉碱+3-羟基丁酰肉碱		戊酰肉碱		己酰肉碱		辛酰肉碱		C4DC+C5OH		癸酰肉碱		十二碳酰肉碱		十四碳酰肉碱		戊二酰肉碱+3-羟基己酰肉碱		十四碳烯酰肉碱		十六碳酰肉碱		十八碳酰肉碱
组别	中位数(X_0.5%, X_99.5%, μmol/L)	变异系数(%)	中位数(X_0.5%, X_99.5%, μmol/L)	变异系数(%)	中位数(X_0.5%, X_99.5%, μmol/L)	变异系数(%)	中位数(X_0.5%, X_99.5%, μmol/L)	变异系数(%)	中位数(X_0.5%, X_99.5%, μmol/L)	变异系数(%)	中位数(X_0.5%, X_99.5%, μmol/L)	变异系数(%)	中位数(X_0.5%, X_99.5%, μmol/L)	变异系数(%)	中位数(X_0.5%, X_99.5%, μmol/L)	变异系数(%)	中位数(X_0.5%, X_99.5%, μmol/L)	变异系数(%)	中位数(X_0.5%, X_99.5%, μmol/L)	变异系数(%)	中位数(X_0.5%, X_99.5%, μmol/L)	变异系数(%)	中位数(X_0.5%, X_99.5%, μmol/L)	变异系数(%)	中位数(X_0.5%, X_99.5%, μmol/L)	变异系数(%)	中位数(X_0.5%, X_99.5%, μmol/L)	变异系数(%)	中位数(X_0.5%, X_99.5%, μmol/L)	变异系数(%)	中位数(X_0.5%, X_99.5%, μmol/L)	变异系数(%)
与室温组比较，^*P < 0.01;与冷链+低温组比较，^#P < 0.01.X_0.5%:0.5%分位数；X_99.5%:99.5%分位数.C4DC+C5OH:甲基丙二酰肉碱+3-羟基异戊酰肉碱.
室温组	23.59(10.38, 58.17)	8.25	14.51(2.98, 40.01)	19.36	1.33(0.42, 4.00)	9.71	0.20(0.10, 0.47)	5.82	0.09(0.03, 0.36)	16.54	0.11(0.05, 0.37)	3.85	0.04(0.02, 0.18)	30.76	0.06(0.02, 0.24)	33.89	0.18(0.08, 0.40)	2.44	0.07(0.03, 0.27)	6.43	0.07(0.03, 0.30)	10.16	0.16(0.06, 0.38)	4.25	0.04(0.01, 0.22)	50.33	0.05(0.03, 0.26)	16.76	2.46(0.53, 6.02)	6.69	0.79(0.26, 1.90)	7.22
冷链+低温组	23.39(10.49, 55.54)^*	5.35	19.14(6.74, 45.39)^*	7.13	1.69(0.55, 4.83)^*	5.25	0.22(0.11, 0.54)^*	4.97	0.12(0.04, 0.41)^*	5.37	0.11(0.05, 0.37)^*	6.34	0.04(0.02, 0.11)^*	10.19	0.06(0.02, 0.18)^*	8.32	0.21(0.10, 0.45)^*	4.22	0.08(0.03, 0.28)^*	4.76	0.08(0.02, 0.34)^*	5.25	0.19(0.07, 0.45)^*	0.00	0.11(0.05, 0.24)^*	4.66	0.06(0.03, 0.30)^*	0.00	3.25(0.72, 7.46)^*	1.53	0.89(0.30, 1.98)^*	1.80
冷链+低温低湿组	20.91(9.79, 49.25)^*#	5.63	19.70(7.34, 46.64)^*#	4.39	1.75(0.58, 4.99)^*#	3.34	0.22(0.11, 0.54)^*#	4.38	0.13(0.04, 0.44)^*#	6.42	0.11(0.05, 0.36)^*#	5.56	0.04(0.02, 0.11)^*#	0.00	0.07(0.03, 0.21)^*#	12.09	0.21(0.10, 0.46)^*#	5.17	0.08(0.03, 0.29)^*#	6.13	0.08(0.02, 0.35)^*#	6.50	0.18(0.06, 0.44)^*#	4.31	0.10(0.04, 0.21)^*#	6.38	0.08(0.03, 0.31)^*#	5.48	3.16(0.66, 7.34)^*#	4.07	0.86(0.27, 1.93)^*#	3.11

表 3 不同递送和储存条件下1 254 616名新生儿干血斑标本中各种肉碱浓度

图 2 不同递送和储存条件下1 254 616名新生儿干血斑标本中各种肉碱浓度月中位数变化趋势图

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