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浙江大学学报（医学版）
浙江大学学报（医学版）  2021, Vol. 50 Issue (4): 487-493    DOI: 10.3724/zdxbyxb-2021-0275
专题报道     
全自动荧光免疫分析仪在新生儿葡萄糖-6-磷酸脱氢酶缺乏症筛查中的应用
缪海霞1(),张玉2,方可欣1,施叶珍2,张婷1,陈荣庆2,吴鼎文1,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
Application of genetic screening processor in screening neonatal glucose-6-phosphate dehydrogenase deficiency
MIAO Haixia1(),ZHANG Yu2,FANG Kexin1,SHI Yezhen2,ZHANG Ting1,CHEN Rongqing2,WU Dingwen1,YANG Rulai1,HUANG Xinwen1,*()
1. Department of Genetics and Metabolism, Children’s Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, National Children’s Regional Medical Center, Hangzhou 310052, China;
2. Zhejiang Biosan Biochemical Technology Co., Ltd., Hangzhou 310012, China
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摘要：

目的：分析全自动荧光免疫分析仪（GSP分析仪）检测新生儿葡萄糖-6-磷酸脱氢酶（G6PD）缺乏症的性能。方法：应用GSP分析仪检测国家卫生健康委临床检验中心室间质量评价质控品和G6PD试剂盒（荧光分析法）的低、高质控品，计算准确度和精密度。采用GSP分析仪和半自动荧光免疫分析仪（1420分析仪）同步检测2622例新生儿筛查标本和41例确诊患儿的标本，分析检测结果的相关性和一致性；采用GSP分析仪和1420分析仪检测漂浮和未漂浮标本各78例，比较检测结果差异；采用1420分析仪和GSP分析仪分别检测2017年1月至2018年12月1?100?384名新生儿筛查标本及2019年1月至2020年12月855?856名新生儿筛查标本，比较两种仪器的筛查效能。对目前使用的切值（26?U/dL）的合理性进行评估，并通过百分位数法，以第99.1百分位建立GSP分析仪筛查G6PD缺乏症的新切值。结果：GSP分析仪检测5个不同浓度室间质控品的检测值与靶值的相对偏倚为0.71%~4.23%，检测G6PD测定试剂盒质控品的批内精密度为4.34%~4.91%，批间精密度为0.85%~2.12%，总变异系数为5.44%~5.72%，均符合实验要求。GSP分析仪和1420分析仪检测的G6PD活性存在明显的正相关（r=0.740，P<0.01），均能筛查出41例确诊患儿，筛查的一致性较好（Kappa=0.945）。1420分析仪检测漂浮干血斑中的G6PD活性明显低于未漂浮干血斑中的G6PD活性（P<0.05），而GSP分析仪检测漂浮和未漂浮干血斑中的G6PD活性差异无统计学意义（P>0.05）。GSP分析仪和1420分析仪筛查G6PD缺乏症的敏感度均为100.00%，且特异度均在99.80%以上。与1420分析仪比较，GSP分析仪筛查G6PD缺乏症的阳性预测值、初筛阳性率和诊断率均上升，假阳性率下降（均P<0.01）。根据人群第99.1百分位建立新切值，男性为26.1?U/dL，女性为29.1?U/dL。结论：GSP分析仪检测性能良好，筛查效率高，更有利于疾病的检出，可用于临床新生儿G6PD缺乏症的大规模筛查。
关键词： 新生儿筛查;  荧光免疫分析;  葡萄糖-6-磷酸脱氢酶缺乏症;  切值    
Abstract:

Objective: To evaluate the performance of genetic screening processor (GSP analyzer) in neonatal screening for glucose-6-phosphate dehydrogenase (G6PD)deficiency. Methods:The accuracy and precision of GSP analyzer was evaluated with the control materials from National Center for Clinical Laboratories and the low and high quality G6PD control kit (fluorescence analysis). GSP analyzer and semi-automatic fluorescence immunoanalyzer (1420 analyzer) were simultaneously used to detect 2622 neonatal screening samples and 41 confirmed samples to analyze the correlation and consistency of the test results; 78 floating samples and 78 non-floating samples were detected to compare the result. A total of 1?100?384 neonatal screening samples from January 2017 to December 2018 and 855?856 neonatal screening samples from January 2019 to December 2020 were detected with 1420 analyzer and GSP analyzer, respectively. Referring to the percentile method and the expert consensus, the new cut-off value of GSP analyzer for G6PD deficiency in screening was established. Results: The relative bias of GSP analyzer in detecting G6PD was 0.71%–4.23%; the intra assay precision was 4.34%–4.91%, the inter assay precision was 0.85%–2.12%, and the total coefficient of variation was 5.44%–5.72%. There was a significant positive correlation between G6PD activity detected by GSP analyzer and 1420 analyzer (r=0.740, P<0.01). Forty-one clinical confirmed patients were identified by both 1420 analyzer and GSP analyzer (Kappa=0.945). The G6PD activity in floating dry blood spots detected by 1420 analyzer was significantly lower than that in non-floating dry blood spots (P<0.05), but there was no significant difference in G6PD activity between floating and non-floating dry blood spots detected by GSP analyzer (P>0.05). The sensitivities of GSP analyzer and 1420 analyzer in screening G6PD deficiency were both 100.00%, and the specificities were both more than 99.80%. Compared with 1420 analyzer, the positive predictive value, positive rate and prevalence of G6PD deficiency detected by GSP analyzer were increased, and the false positive rate was decreased (allP<0.01). The new cut-off value was 26.1 U/dL for male and 29.1 U/dL for female according to the 99.1% percentile of the population.Conclusion: GSP analyzer has better detection performance with high automation, efficiency and throughput, which can be used in large-scale screening for neonatal G6PD deficiency.
Key words: Neonatal screening    Fluorescence immunoassay    Glucose-6-phosphate dehydrogenase deficiency    Cut-off value
收稿日期: 2021-04-09 出版日期: 2021-11-01
CLC:  R446.1  
基金资助: 国家重点研发计划（2018YFC1002204）
通讯作者: 黄新文     E-mail: hamhx@zju.edu.cn;6305022@zju.edu.cn
作者简介: 缪海霞，主管技师，主要从事儿童遗传代谢病筛查工作；E-mail：hamhx@zju.edu.cn；https：//orcid.org/0000-0001-9400-1810
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缪海霞
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引用本文:

缪海霞,张玉,方可欣,施叶珍,张婷,陈荣庆,吴鼎文,杨茹莱,黄新文. 全自动荧光免疫分析仪在新生儿葡萄糖-6-磷酸脱氢酶缺乏症筛查中的应用[J]. 浙江大学学报（医学版）, 2021, 50(4): 487-493.

MIAO Haixia,ZHANG Yu,FANG Kexin,SHI Yezhen,ZHANG Ting,CHEN Rongqing,WU Dingwen,YANG Rulai,HUANG Xinwen. Application of genetic screening processor in screening neonatal glucose-6-phosphate dehydrogenase deficiency. J Zhejiang Univ (Med Sci), 2021, 50(4): 487-493.

链接本文:

http://www.zjujournals.com/med/CN/10.3724/zdxbyxb-2021-0275    或    http://www.zjujournals.com/med/CN/Y2021/V50/I4/487

GSP分析仪

1420分析仪

合计

阳性

阴性

阳性

53

2

55

阴性

4

2604

2608

合计

57

2606

2663
表 1  GSP分析仪与1420分析仪筛查G6PD缺乏症的一致性分析

筛查方法

n

初筛阳性例数

初筛阳性率（%）

诊断例数

诊断率（%）

假阳性率（%）

阳性预测值（%）

敏感度（%）

特异度（%）

1420分析仪

?

男性

579?993

5104

0.88

3973

0.69

0.20

77.84

100.00

99.80

女性

520?391

1349

0.26

501

0.10

0.18

37.14

100.00

99.84

合计

1?100?384

6453

0.59

4474

0.41

0.18

69.33

100.00

99.82

GSP分析仪

?

男性

449?595

4017

0.89

3340

0.74**

0.15**

83.15**

100.00

99.85

女性

406?261

1378

0.34**

630

0.16**

0.16**

45.72**

100.00

99.82

合计

855?856

5395

0.63**

3970

0.46**

0.17**

73.59**

100.00

99.83
表 2  GSP分析仪与1420分析仪对G6PD缺乏症的筛查效率比较
图 1  GSP分析仪检测男性和女性标本中G6PD活性的分布A：男性标本筛查结果；B：女性标本筛查结果. G6PD：葡萄糖-6-磷酸脱氢酶.
1 BEUTLERE. Glucose-6-phosphate dehydrogenase deficiency: a historical perspective[J]Blood, 2008, 111( 1): 16-24.
doi: 10.1182/blood-2007-04-077412
2 中华人民共和国卫生部. 新生儿疾病筛查技术规范(2010年版)[A/OL]. (2010-11-10)[2020-06-11]. http://www.nhc.gov.cn/cmsresources/mohfybjysqwss/cmsrsdocument/doc10798.doc
Ministry of Health of the People’s Republic of China. Technical guide of newborn screening in China (2010) [A/OL]. (2010-11-10) [2020-06-11]. http://www.nhc.gov.cn/cmsresources/mohfybjysqwss/cmsrsdocument/doc10798.doc. (in Chinese)
3 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, 中国医师协会医学遗传医师分会临床生化遗传专业委员会, 中国医师协会青春期医学专业委员会临床遗传学组. 葡萄糖-6-磷酸脱氢酶缺乏症新生儿筛查、诊断和治疗专家共识[J]. 中华儿科杂志, 2017, 55(6): 411-414
The Subspecialty Group of Newborn Screening, Society of Birth Defects Prevention and Control, Chinese Preventive Medicine Association, The Subspecialty Committee of Clinical Biochemical Genetics, Society of Medical Genetics, Chinese Medical Doctor Association, The Subspecialty Group of Clinical Genetics, Society of Adolescent Medicine, Chinese Medical Doctor Association. Expert consensus on neonatal screening, diagnosis and treatment of glucose-6-phosphate dehydrogenasedeficiency[J]. Chinese Journal of Pediatrics, 2017, 55(6): 411-414. (in Chinese)
4 Clinical and Laboratory Standards Institute. EP15-A2. User verification of performance for precision and trueness; Approved Guideline-Second Edition[S/OL]. [2021-02-01]. https://webstore.ansi.org/standards/clsi/clsiep15a2
5 Clinical and Laboratory Standards Institute. EP05-A2. Evaluation of precision performance of quantitative measurement methods; approved guideline-second edition[S/OL]. [2021-02-01]. https://webstore.ansi.org/standards/clsi/clsiep05a2?gclid=EAIaIQobChMIgdebuPfO8gIVAcuWCh2VfgzlEAAYASAAEgJOofD_BwE
6 Clinical and Laboratory Standards Institute. How to define and determine reference intervals in the clinical laboratory. CLSI C28?A2 Ed. 2[S]. Wayne, PA: CLSI, 2000
7 国家卫生健康委临床检验中心新生儿疾病筛查室间质评专家委员会. 新生儿葡萄糖-6-磷酸脱氢酶缺乏症筛查与诊断实验室检测技术专家共识[J]. 中华检验医学杂志, 2019, 42(3): 181-185
Expert Committee for Inter Room Quality Evaluation of Neonatal Disease Screening in Clinical Laboratory Center of National Health Commission. Consensus of experts on detection technology in neonatal glucose-6-phosphate dehydrogenase deficiency screening and diagnosis laboratory[J].Chinese Journal of Laboratory Medicine, 2019, 42 (3): 181-185. (in Chinese)
8 FINGERHUTR, TORRESANIT. Evaluation of the genetic screening processor (GSP) for newborn screening[J]Anal Methods, 2013, 5( 18): 4769.
doi: 10.1039/c3ay40593a
9 DE CASTROS M, FILIPPONL, SOUZAA C, et al.Evaluation of the genetic screening processor for the performance of newborn screening tests[J]J Inborn Errors Metab Screening, 2016, 1-5.
doi: 10.1177/2326409816669370
10 梁晓威, 蒋　涛, 程　威, 等. 全自动免疫荧光分析仪检测苯丙氨酸的性能验证[J]. 南京医科大学学报(自然科学版), 2018, 38(5): 699-702
LIANG Xiaowei, JIANG Tao, CHENG Wei, et al. Performance verification of automatic immunofluorescence analyzer for the detection of phenylalanine[J]. Journal of Nanjing Medical University (Natural Science Edition), 2018, 38(5): 699-702. (in Chinese)
11 田国力, 王燕敏, 周　卓, 等. 全自动新生儿疾病筛查系统的性能评价[J]. 检验医学, 2017, 32(7): 633-636
TIAN Guoli, WANG Yanmin, ZHOU Zhuo, et al. Performance evaluation of automatic neonatal disease screening system[J]. Laboratory Medicine, 2017, 32(7): 633-636. (in Chinese)
12 LEYB, ALAMM S, O’DONNELLJ J, et al.A comparison of three quantitative methods to estimate G6PD activity in the chittagong hill tracts, bangladesh[J/OL]PLoS One, 2017, 12( 1): e0169930.
doi: 10.1371/journal.pone.0169930
13 ANANTASOMBOONP, CHANDAM, JUGNAM-ANGW, et al.Evaluating the performance of automated UV enzymatic assay for screening of glucose 6‐phosphate dehydrogenase deficiency[J]Int J Lab Hematol, 2019, 41( 2): 192-199.
doi: 10.1111/ijlh.12943
14 FLORESS R, HALLE M, DE JESúSV R. Glucose-6-phosphate dehydrogenase enzyme stability in filter paper dried blood spots[J]Clin Biochem, 2017, 50( 15): 878-881.
doi: 10.1016/j.clinbiochem.2017.05.003
15 ALAMM S, KIBRIAM G, JAHANN, et al.Field evaluation of quantitative point of care diagnostics to measure glucose-6-phosphate dehydrogenase activity[J/OL]PLoS One, 2018, 13( 11): e0206331.
doi: 10.1371/journal.pone.0206331
16 JIANGW Y, YUG L, LIUP, et al.Structure and function of glucose-6-phosphate dehydrogenase-deficient variants in Chinese population[J]Hum Genet, 2006, 119( 5): 463-478..
doi: 10.1007/s00439-005-0126-5
17 KAPLANM, BEUTLERE, VREMANH J, et al.Neonatal hyperbilirubinemia in glucose-6-phosphate dehydrogenase-deficient heterozygotes[J]Pediatrics, 1999, 104( 1): 68-74.
doi: 10.1542/peds.104.1.68
18 KAPLAN M, HAMMERMAN C, VREMAN H J. Acute haemolysis and severe neonatal hyper-bilirubinemia in glucose-6-phosphate dehydrogenase deficient heterozygote[J]. J Pediatr, 2001, 139(1): 137-140
19 AINOON O, ALAWIYAH A, YU Y H, et al. Semiquantitative screening test for G6PD deficiency detects severe deficiency but misses a substantial proportion of partially-deficient females[J]. Southeast Asian J Trop Med Public Health, 2003, 34(2): 405-414
20 FUC, LUOS, LIQ, et al.Newborn screening of glucose-6-phosphate dehydrogenase deficiency in Guangxi, China: determination of optimal cutoff value to identify heterozygous female neonates[J]Sci Rep, 2018, 8( 1): 833.
doi: 10.1038/s41598-017-17667-6
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