化学发光法检测半乳糖凝集素3的性能评价及其在心力衰竭中的初步应用

doi:10.3969/j.issn.1673-8640.2019.04.016

摘要/Abstract

摘要：

目的评价化学发光法检测半乳糖凝集素3（Gal-3）的分析性能并初步探讨其对心力衰竭（HF）的诊断价值。方法依据相关文件评价化学发光法检测Gal-3的分析性能[空白检测限（LoB）、检出限（LoD）、定量限（LoQ）、精密度、线性范围及抗干扰能力]并验证厂家声明的参考区间,比较年龄、性别和样本类型对化学发光法测定Gal-3的影响。检测55例HF患者（HF组）及80名体检健康者（正常对照组）的血浆Gal-3水平,采用受试者工作特征（ROC）曲线评估Gal-3诊断HF的价值。结果 Gal-3试剂（化学发光法）生产厂家声明的LoB、LoD、LoQ分别为0.80、1.0、1.0 ng/mL,验证均通过。重复精密度[以变异系数（CV）表示]为1.9%~4.8%、期间精密度（以CV表示）为2.6%~6.1%,精密度验证通过。在4.07~113.11 ng/mL范围内,化学发光法测定Gal-3呈线性。血红蛋白<20 g/L、总胆红素（TB）<403.2 μmol/L、直接胆红素（DBil）<214.08 μmol/L、乳糜浊度<7 450 FTU对化学发光法检测血清Gal-3无干扰。血浆的Gal-3测定值高于血清（P<0.05）。男、女性之间Gal-3水平差异无统计学意义（P>0.05）。将研究对象按样本类型（血清、血浆）分组后再按年龄分组,血清组中3个年龄亚组（20~<45岁组、45~<60岁组、≥60岁组）之间Gal-3水平差异有统计学意义（P<0.05）,血浆组2个年龄亚组（20~<60岁组、≥60岁组）之间Gal-3水平差异有统计学意义（P<0.05）。Gal-3水平随年龄增高而升高。HF患者血浆Gal-3水平明显高于正常对照者（P<0.001）。ROC曲线分析显示,血浆Gal-3诊断HF的曲线下面积（AUC）为0.969,最佳临界值为20.04 ng/mL,敏感性为85.5%,特异性为100.0%。结论化学发光法检测Gal-3的分析性能良好,需注意的是年龄对Gal-3水平有一定的影响。Gal-3对HF有较高的辅助诊断价值。

关键词: 半乳糖凝集素3, 化学发光法, 分析性能, 心力衰竭

Abstract:

Objective To evaluate the performance of galectin-3（Gal-3） detection by chemiluminescence and its preliminary application in heart failure（HF）. Methods According to related documents,the performance of Gal-3 detection by chemiluminescence [limit of blank（LoB）,limit of detection（LoD）,limit of quantitation（LoQ）,precision,linear range and interference] was evaluated,and the manufacturer's reference interval was verified. The effects of age,sex and specimen type on Gal-3 detection were compared. A total of 55 patients with HF（HF group） and 80 healthy subjects（healthy control group） were enrolled,and their plasma samples were collected for Gal-3 detection. Receiver operating characteristic（ROC） curve was used to analyze the role of Gal-3 in HF diagnosis. Results The Gal-3 kit（chemiluminescence） got a satisfying result in verification on LoB,LoD and LoQ,with the values of 0.80,1.0 and 1.0 ng/mL by the manufacturer,respectively. The repeat precision expressed by coefficient of variation（CV） was 1.9%-4.8%,and the within-laboratory precision expressed by CV was 2.6%-6.1%,and the manufacturer's precision got verification. In the range of 4.07-113.11 ng/mL,there was a good linearity of Gal-3 detection by chemiluminescence. Serum Gal-3 level determined by chemiluminescence would not be affected in samples with hemoglobin level <20 g/L,total bilirubin（TB）<403.2 μmol/L,direct bilirubin（DBil） < 214.08 μmol/L or the turbidity of chyle <7 450 FTU. The plasma Gal-3 level was higher than that in serum （P<0.05）. There was no statistical significance in Gal-3 levels between males and females（P>0.05）. The research subjects were classified by sample types（serum and plasma） and ages. In serum group,Gal-3 level had statistical significance in 3 age subgroups（20~<45-year-old group,45~<60-year-old group and ≥60-year-old group）（P<0.05）,and there was statistical significance for plasma Gal-3 levels in 20~<60-year-old group and ≥60-year-old group（P<0.05）. Gal-3 levels increased with ages. The level of Gal-3 was higher in patients with HF than that in healthy control group（P<0.001）. The area under ROC curve （AUC） of Gal-3 for HF diagnosis was 0.969,and the optimal cut-off value was 20.04 ng/mL,with a sensitivity of 85.5% and a specificity of 100.0%. Conclusions The analytical performance of Gal-3 detection by chemiluminescence is good. Age has a certain effect on Gal-3 levels. Gal-3 plays a role in the auxiliary diagnosis of heart failure.

Key words: Galectin-3, Chemiluminescence, Analytical performance, Heart failure

中图分类号:

R446.1

郑笑荣, 万军, 张良颖, 李永姝, 韩雪晶, 石萍, 贾克刚. 化学发光法检测半乳糖凝集素3的性能评价及其在心力衰竭中的初步应用[J]. 检验医学, 2019, 34(4): 360-366.

ZHENG Xiaorong, WAN Jun, ZHANG Liangying, LI Yongshu, HAN Xuejing, SHI Ping, JIA Kegang. Performance evaluation of galectin-3 detection by chemiluminescence and its preliminary application in heart failure[J]. Laboratory Medicine, 2019, 34(4): 360-366.

图/表 7

参考文献 23

[1]	原永明,王慧,尹良平,等. 慢性心力衰竭患者BNP、hs-CRP水平检测的临床意义[J]. 检验医学,2015,30(6):591-592.
[2]	FILIPE M D,MEIJERS W C,ROGIER VAN DER VELDE A,et al. Galectin-3 and heart failure:prognosis,prediction and clinical utility[J]. Clin Chim Acta,2015,443:48-56.
[3]	贾克刚. 心力衰竭的新兴标志物-半乳糖凝集3[J]. 中华检验医学杂志,2013,36(6):506-510.
[4]	SCHINDLER E I,SZYMANSKI J J,HOCK K G,et al.Short- and long-term biologic variability of galectin-3 and other cardiac biomarkers in patients with stable heart failure and healthy adults[J]. Clin Chem,2016,62(2):360-366.
[5]	YANCY C W,JESSUP M,BOZKURT B,et al.2013 ACCF/AHA guideline for the management of heart failure:executive summary:a report of the American College of Cardiology Foundation/American Heart Association Task Force on practice guidelines[J]. Circulation,2013,128(16):1810-1852.
[6]	U. S. Food and Drug Administration. Device market approval under the federal food,drug and cosmetic act. 21 CFR 862.1117[R]. New York:FDA,2010.
[7]	中华医学会心血管病学分会,中华心血管病杂志编辑委员会. 中国心力衰竭诊断和治疗指南2014[J]. 中华心血管病杂志,2014,42(2):98-122.
[8]	中国合格评定国家认可委员会. CNAS-CL02 医学实验室质量和能力认可准则[S]. 中国:中国合格评定国家认可委员会,2013.
[9]	Clinical and Laboratory Standards Institute. Protocols for determination of limits of detection and limits of quantitation[S]. EP17-A,CLSI,2004.
[10]	Clinical and Laboratory Standards Institute. Evaluation of precision performance of clinical chemistry devices[S]. EP5-A2,CLSI,2004.
[11]	Clinical and Laboratory Standards Institute. Evaluation of the linearity of quantitative measurement procedures[S]. EP6-A,CLSI,2003.
[12]	Clinical and Laboratory Standards Institute. Interference testing in clinical chemistry[S]. EP7-A2,CLSI,2005.
[13]	Clinical and Laboratory Standards Institute. How to define and determine reference intervals in the clinical laboratory[S]. C28-A2,CLSI,2000.
[14]	PONIKOWSKI P,VOORS A A,ANKER S D,et al.2016 ESC Guidelines for the diagnosis and treatment of acute and chronic heart failure:the task force for the diagnosis and treatment of acute and chronic heart failure of the European Society of Cardiology(ESC) developed with the special contribution of the Heart Failure Association(HFA) of the ESC[J]. Eur Heart J,2016,37(27):2129-2200.
[15]	LI L C,LI J,GAO J.Functions of galectin-3 and its role in fibrotic diseases[J]. J Pharmacol Exp Ther,2014,351(2):336-343.
[16]	MACKINNON A C,FARNWORTH S L,HODKINSON P S,et al.Regulation of alternative macrophage activation by galectin-3[J]. J Immunol, 2008,180(4):2650-2658.
[17]	SHARMA U C,POKHAREL S,VAN BRAKEL T J,et al. Galectin-3 marks activated macrophages in failure-prone hypertrophied hearts and contributes to cardiac dysfunction[J]. Circulation,2004,110(19):3121-3128.
[18]	CALVIER L,MARTINEZ-MARTINEZ E,MIANA M,et al.The impact of galectin-3 inhibition on aldosterone-induced cardiac and renal injuries[J]. JACC Heart Fail,2015,3(1):59-67.
[19]	CALVIER L,MIANA M,REBOUL P,et al.Galectin-3 mediates aldosterone-induced vascular fibrosis[J]. Arterioscler Thromb Vasc Biol,2013,33(1):67-75.
[20]	GAZE D C,PRANTE C,DREIER J,et al.Analytical evaluation of the automated galectin-3 assay on the Abbott ARCHITECT immunoassay instruments[J]. Clin Chem Lab Med,2014,52(6):919-926.
[21]	KRINTUS M,KOZINSKI M,FABISZAK T,et al.Establishing reference intervals for galectin-3 concentrations in serum requires careful consideration of its biological determinants[J]. Clin Biochem,2017,50(10-11):599-604.
[22]	MEIJERS W C,VAN DER VELDE A R,DE BOER R A. The ARCHITECT galectin-3 assay:comparison with other automated and manual assays for the measurement of circulating galectin-3 levels in heart failure[J]. Expert Rev Mol Diagn,2014,14(3):257-266.
[23]	YU L,RUIFROK W P,MEISSNER M,et al.Genetic and pharmacological inhibition of galectin-3 prevents cardiac remodeling by interfering with myocardial fibrogenesis[J]. Circ Heart Fail,2013,6(1):107-117.

质控品	标示值（ng/mL）	重复精密度			期间精密度
质控品	标示值（ng/mL）	s_r（ ng/mL）	CV（%）	σ_r （ ng/mL）	s_l（ ng/mL）	CV（%）	σ_l（ ng/mL）
低值	9.1	0.31	4.8	0.44	0.42	6.1	0.56
中值	20.5	0.38	2.5	0.51	0.52	3.4	0.70
高值	74.1	1.22	1.9	1.41	1.69	2.6	1.93

质控品	标示值（ng/mL）	重复精密度			期间精密度
质控品	标示值（ng/mL）	s_r（ ng/mL）	CV（%）	σ_r （ ng/mL）	s_l（ ng/mL）	CV（%）	σ_l（ ng/mL）
低值	9.1	0.31	4.8	0.44	0.42	6.1	0.56
中值	20.5	0.38	2.5	0.51	0.52	3.4	0.70
高值	74.1	1.22	1.9	1.41	1.69	2.6	1.93

理论值（ng/mL）	测定值1 （ng/mL）	测定值2 （ng/mL）	x （ng/mL）	百分差值（%）
4.00	4.07	4.06	4.065	0.246
26.00	22.19	22.98	22.585	3.498
48.00	40.76	41.75	41.255	2.400
70.00	63.56	65.74	64.650	3.372
92.00	82.60	78.98	80.790	4.481
114.00	113.81	112.40	113.105	1.247

理论值（ng/mL）	测定值1 （ng/mL）	测定值2 （ng/mL）	x （ng/mL）	百分差值（%）
4.00	4.07	4.06	4.065	0.246
26.00	22.19	22.98	22.585	3.498
48.00	40.76	41.75	41.255	2.400
70.00	63.56	65.74	64.650	3.372
92.00	82.60	78.98	80.790	4.481
114.00	113.81	112.40	113.105	1.247

干扰物	Gal-3		相对偏移（%）
干扰物	初始值	测定值	相对偏移（%）
血红蛋白（g/L）
20	7.36	7.99	8.6
25	6.13	7.79	27.1
TB（µmol/L）
403.2	4.68	5.22	11.5
470.4	3.51	4.39	25.1
DBil（µmol/L）
214.08	4.68	5.22	11.5
249.76	3.51	4.39	25.1
乳糜浊度（FTU）
7 450	5.79	6.57	13.5
8 940	4.63	5.58	20.5