CN113699211A - 一种用于儿童威尔森氏症快速筛查的参数组合和试剂盒 - Google Patents

一种用于儿童威尔森氏症快速筛查的参数组合和试剂盒 Download PDF

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CN113699211A
CN113699211A CN202110821325.6A CN202110821325A CN113699211A CN 113699211 A CN113699211 A CN 113699211A CN 202110821325 A CN202110821325 A CN 202110821325A CN 113699211 A CN113699211 A CN 113699211A
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周建利
周少明
陈沫先
赵煜桢
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Abstract

本发明提供了一种用于儿童威尔森氏症快速筛查的参数组合和试剂盒。该参数组合包括ALT水平、血浆铜蓝蛋白水平和24小时尿铜水平中一种或者多种。试剂盒，其包括检测ALT水平、血浆铜蓝蛋白水平和/或尿铜水平的试剂；和一说明书。本发明提供的参数组合物和试剂盒通过参考ALT升高、血浆铜蓝蛋白水平降低和24小时尿铜升高三个参数，可以适用于威尔森氏症的早期诊断和筛查，操作简单易行，可信度高，便于早期的诊断和介入治疗，将有利于大大提高生存率并具有良好的预后。

Description

一种用于儿童威尔森氏症快速筛查的参数组合和试剂盒
技术领域
本发明涉及医学检测技术领域，具体涉及一种用于儿童威尔森氏症快速筛查的参数组合和试剂盒。
背景技术
肝豆状核变性(HLD)，也称为威尔逊病/威尔森氏症(WD)，是一种常染色体隐性遗传病。该病在世界各地均有发生，在人群中的发病率约为活产儿的1/30,000[1]。WD是由编码13号染色体上细胞内铜转运蛋白的ATP7B基因突变导致的，导致细胞内铜输出受损[2]。ATP7B是一种P型ATP酶，主要在肝脏中表达。ATP7B将铜与其N端结构域结合，并负责铜跨膜运输，使用ATP作为其能源。研究表明，不同位点的突变会影响ATPase的活性。迄今为止，已在WD患者中发现ATP7B位点的800多种不同突变[3]。特别是，WD ATP7B基因的突变影响铜离子与铜蓝蛋白的相互作用，进而影响胆汁中铜的排泄，是肝脏铜排泄的主要途径。如果胆汁中的铜排泄减少，铜就会沉积在肝脏周围的地方，造成肝细胞损伤，临床上首先诊断为丙氨酸氨基转移酶(ALT)升高。随后铜在脑、角膜和肾脏上的沉积会对相应器官造成损害，并伴有各种临床症状[4]。随着时间的推移，肝脏逐渐受到沉积铜的损害，部分患者最终会出现肝硬化或肝功能衰竭，以及严重的神经系统损害、血液系统损害和精神症状[4]。
由于治疗开始的时间不同，WD儿童的临床表现可能多种多样[5,6]。通常，新生婴儿的铜排泄机制尚未完全发育，在出生后一年内变得更加有效。然而，WD患者铜排泄的关键途径未能发展或出现功能障碍，导致患者一生中铜的积累，逐渐产生各种临床症状[7]。一般来说，虽然WD患者的诊断年龄为5-35岁(平均＝13岁)[8]，但也有年轻和年龄较大的患者(>70)需要诊断[4,8-10]。此外，一项对143名WD儿童的研究表明，21名儿童(15％)在5岁前出现肝功能异常的症状或检查结果[10]。WD儿童平均年龄在9-13岁最常见的初始表现是肝脏疾病[11,12]，而WD占慢性活动性肝炎儿童的8％-10％[13]。对于其他患者，肝病可能无症状进展，肝硬化或急性肝功能衰竭的多种并发症往往直到青春期才会出现。此外，在某些情况下，在明显的肝脏疾病之前也可能出现神经或精神表现。总而言之，儿童和老年患者更可能分别被诊断为肝脏和神经系统表现。老年患者出现神经系统症状的平均年龄为15-21岁[6,11,14,15]。WD发病年龄的差异可能反映了基因突变和外显率、基因外因素和其他环境因素(例如饮食)的变化[16]。现有技术主要依据基因突变检测分析，病人多半鉴于病情出现表征后才会进行检测，不适合早期体检筛查中使用。因此，高精度的早期诊断对于WD患者及其预后至关重要。
发明内容
为了克服现有技术中的缺陷，本发明提供了一种用于儿童威尔森氏症快速筛查的参数组合和试剂盒。
为实现上述目的，本发明采用如下技术方案：
本发明的第一方面是提供一种用于儿童威尔森氏症快速筛查的参数组合，包括ALT水平、血浆铜蓝蛋白水平和24小时尿铜水平中一种或者多种。
进一步地，ALT升高、血浆铜蓝蛋白水平降低和/或24小时尿铜升高，则可以初步诊断为威尔森氏症。
本发明的第二方面是提供一种用于儿童威尔森氏症快速筛查的试剂盒，其包括检测ALT水平、血浆铜蓝蛋白水平和/或尿铜水平的试剂；和
一说明书。
进一步地，上述说明书包括如下指示：
a.检测ALT水平、血浆铜蓝蛋白水平和24小时尿铜水平；和
b.判断是否可以初步诊断为威尔森氏症：若ALT升高、血浆铜蓝蛋白水平降低和/或24小时尿铜升高，则可以初步诊断为威尔森氏症；若无任一符合上述标准的指标，则可以判断未患有威尔森氏症。
进一步地，上述说明书还包括如下指示：
c.若可以初步诊断为威尔森氏症，则再采用ATP7B基因的基因突变试验用于确诊是否为威尔森氏症。
本发明采用以上技术方案，与现有技术相比，具有如下技术效果：
本发明提供的参数组合物和试剂盒通过参考ALT升高、血浆铜蓝蛋白水平降低和24小时尿铜升高三个参数，可以适用于威尔森氏症的早期诊断和筛查，操作简单易行，可信度高，便于早期的诊断和介入治疗，将有利于大大提高生存率并具有良好的预后。
具体实施方式
本发明提供了一种用于儿童威尔森氏症快速筛查的参数组合和试剂盒。该参数组合包括ALT水平、血浆铜蓝蛋白水平和24小时尿铜水平中一种或者多种。ALT升高、血浆铜蓝蛋白水平降低和/或24小时尿铜升高，则可以初步诊断为威尔森氏症。
下面通过具体实施例对本发明进行详细和具体的介绍，以使更好的理解本发明，但是下述实施例并不限制本发明范围。
实施例中方法如无特殊说明的采用常规方法，使用的试剂如无特殊说明的使用常规市售试剂或按常规方法配制的试剂。
实施例1
本实施例验证包括ALT水平、血浆铜蓝蛋白水平和24小时尿铜水平中一种或者多种的参数组合可以适用于威尔森氏症的早期快速筛查，具体的操作过程和结果如下：
招募了深圳市儿童医院就诊的30名以丙氨酸氨基转移酶ALT升高为WD首发表现的华南地区儿童，其中女14例，男16例，并对临床资料和基因检测结果的横断面进行了回顾性分析及其短期预后的随访研究。
就诊期间，上述30名儿童患者被进行了体检，发现肝酶升高30例(100％)，且在所有30例(100％)中，血清铜蓝蛋白(CP)水平降低，而24小时尿铜排泄量增加(如下表1)。
表1 30名儿童患者的临床资料
Figure BDA0003172047840000041
Figure BDA0003172047840000051
注：Hb：血红蛋白，ALT：谷丙转氨酶，AST：谷草转氨酶，TB：总胆红素，ALP：血清碱性磷酸酶，GGT：γ-谷氨酰转肽酶，CP：血清铜蓝蛋白。
最终，使用经典确诊方法，ATP7B基因的基因突变试验用于确诊，其中c.2333G>T和c.3443T>C这两个突变位点的突变频率最高，分别占突变位点总数的23.0％和10.7％(如下表2、3)。
表2 30名儿童患者ATP7B的突变类型
Figure BDA0003172047840000052
Figure BDA0003172047840000061
表3 30名儿童患者中的ATP7B突变类型总结
Figure BDA0003172047840000071
Figure BDA0003172047840000081
实施例2
本实施例提供了一种用于儿童威尔森氏症快速筛查的试剂盒，其包括检测ALT水平、血浆铜蓝蛋白水平和/或尿铜水平的试剂；和一说明书。
该说明书包括包括如下指示：
a.检测ALT水平、血浆铜蓝蛋白水平和24小时尿铜水平；和
b.判断是否可以初步诊断为威尔森氏症：若ALT升高、血浆铜蓝蛋白水平降低和24小时尿铜升高，则可以初步诊断为威尔森氏症；若不符合上述标准的指标，则可以判断未患有威尔森氏症；
c.若可以初步诊断为威尔森氏症，则再采用ATP7B基因的基因突变试验用于确诊是否为威尔森氏症。
由上述内容可知，本发明提供的试剂盒便于较早地诊断出威尔森氏症，适用于常规体格检查，且适用于头颅MRI检查阴性和无神经系统表现的患者筛查。
以上对本发明的具体实施例进行了详细描述，但其只作为范例，本发明并不限制于以上描述的具体实施例。对于本领域技术人员而言，任何对本发明进行等同修改和替代也都在本发明的范畴之中。因此，在不脱离本发明精神和范围下所作的均等变换和修改，都应涵盖在本发明的范围内。
参考文献
[1]Huster D.Wilson disease.Best Pract Res Clin Gastroenterol,2010；24:531.
[2]Meranthi F,Indra VM,Evangeline W,et al.Wilson disease in childrenand adolescents.Arch Dis Child,2020；105(5):499-505.
[3]Chen YC,Yu H,Wang RM,et al.Contribution of intragenic deletions tomutation spectrum in Chinese patients with Wilson's disease and possiblemechanism underlying ATP7B gross deletions.Parkinsonism RelatDisord.2019；62:128-133.
[4]European Association for Study of Liver.EASL Clinical PracticeGuidelines:Wilson's disease.J Hepatol 2012；56:671.
[5]Moores A,Fox S,Lang A,et al.Wilson disease:Canadian perspectiveson presentation and outcomes from an adult ambulatory setting.Can JGastroenterol 2012；26:333.
[6]Lorincz MT.Neurologic Wilson's disease.Ann N Y Acad Sci 2010；1184:173.
[7]Manolaki N,Nikolopoulou G,Daikos GL,et al.Wilson disease inchildren:analysis of 57cases.J Pediatr Gastroenterol Nutr 2009；48:72.
[8]Lin LJ,Wang DX,Ding NN,et al.Comprehensive analysis on clinicalfeatures of Wilson's disease:an experience over 28 years with 133cases.Neurol Res 2014；36:157.
[9]Stremmel W,Meyerrose KW,Niederau C,et al.Wilson disease:clinicalpresentation,treatment,and survival.Ann Intern Med 1991；115:720.
[10]Wiernicka A,
Figure BDA0003172047840000091
M,Jańczyk W,et al.Early Onset of WilsonDisease:Diagnostic Challenges.J Pediatr Gastroenterol Nutr 2017；65:555.
[11]Saito T.Presenting symptoms and natural history of Wilsondisease.Eur J Pediatr 1987；146:261.
[12]Walshe JM.Wilson's disease presenting with features of hepaticdysfunction:a clinical analysis of eighty-seven patients.Q J Med 1989；70:253.
[13]Gitlin JD.Wilson disease.Gastroenterology 2003；125:1868.
[14]Oder W,Grimm G,Kollegger H,et al.Neurological andneuropsychiatric spectrum of Wilson's disease:a prospective study of 45cases.J Neurol 1991；238:281.
[15]Ferenci P,
Figure BDA0003172047840000092
A,Merle U,et al.Late-onset Wilson'sdisease.Gastroenterology 2007；132:1294.
[16]Wiggelinkhuizen M,Tilanus ME,Bollen CW,Houwen RH.Systematicreview:clinical efficacy of chelator agents and zinc in the initial treatmentof Wilson disease.Aliment PharmacolTher 2009；29:947.

Claims (5)

1.一种用于儿童威尔森氏症快速筛查的参数组合，其特征在于，包括ALT水平、血浆铜蓝蛋白水平和24小时尿铜水平中一种或者多种。
2.根据权利要求1所述的参数组合，其特征在于，ALT升高、血浆铜蓝蛋白水平降低和/或24小时尿铜升高，则可以初步诊断为威尔森氏症。
3.一种用于儿童威尔森氏症快速筛查的试剂盒，其特征在于，包括检测ALT水平、血浆铜蓝蛋白水平和/或尿铜水平的试剂；和一说明书。
4.根据权利要求3所述的试剂盒，其特征在于，所述说明书包括如下指示：
a.检测ALT水平、血浆铜蓝蛋白水平和24小时尿铜水平；和
b.判断是否可以初步诊断为威尔森氏症：若ALT升高、血浆铜蓝蛋白水平降低和/或24小时尿铜升高，则可以初步诊断为威尔森氏症；若无任一符合上述标准的指标，则可以判断未患有威尔森氏症。
5.根据权利要求4所述的试剂盒，其特征在于，所述说明书还包括如下指示：
c.若可以初步诊断为威尔森氏症，则再采用ATP7B基因的基因突变试验用于确诊是否为威尔森氏症。
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115216533A (zh) * 2022-06-30 2022-10-21 湖南家辉生物技术有限公司 一种用于诊断威尔逊病的生物标志物、扩增引物组、检测试剂及应用

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1379247A (zh) * 2001-03-31 2002-11-13 智诺百股份有限公司 测定血斑中血浆铜蓝蛋白浓度的方法及肝豆状核变性病筛选试剂盒和诊断试剂
WO2020102723A1 (en) * 2018-11-16 2020-05-22 Encoded Therapeutics, Inc. Compositions and methods for treating wilson's disease

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1379247A (zh) * 2001-03-31 2002-11-13 智诺百股份有限公司 测定血斑中血浆铜蓝蛋白浓度的方法及肝豆状核变性病筛选试剂盒和诊断试剂
WO2020102723A1 (en) * 2018-11-16 2020-05-22 Encoded Therapeutics, Inc. Compositions and methods for treating wilson's disease

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
陆怡 等: "儿童肝豆状核变性临床表型及ATP7B基因突变关联性分析", 《中国循证儿科杂志》, vol. 8, no. 5, pages 346 - 351 *
陈冬梅 等: "小儿肝豆状核变性41例临床分析", 《现代生物医学进展》, vol. 9, no. 12, pages 2313 - 2315 *
黄榕 等: "肝豆状核变性33例的临床特点分析", 《现代消化及介入诊疗》, vol. 16, no. 5, pages 328 - 330 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115216533A (zh) * 2022-06-30 2022-10-21 湖南家辉生物技术有限公司 一种用于诊断威尔逊病的生物标志物、扩增引物组、检测试剂及应用

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