Establishment and performance evaluation of chemiluminescent immunoassay for serum HER2 protein

doi:10.3969/j.issn.1000-1565.2022.03.009

Abstract

Abstract: This study established magnetic particles chemiluminescence method for the detection of serum human epidermal growth factor receptor 2(HER2)protein in the patients with breast cancer, and evaluated its performance and equivalence with market products. The detection range of HER2 protein was from 0 to 300 ng/mL; limit of blank(LoB)was 0.48 ng/mL; average recovery rates of exogenous and endogenous interference was lower than 5%; precision was coefficient of variation(CV)lower than 8%; the stability test showed that the reagent could be stored stably for 48 weeks, and the correlation coefficient with commercial products was r=0.995, the mean bias was -0.2 ng/mL. These results indicated that the detection method of serum HER2 protein was successfully established, and had an excellent- DOI:10.3969/j.issn.1000-1565.2022.03.009血清HER2蛋白化学发光免疫检测方法的建立及评价王娟^1,2,程华³,胡琛光^4,5,林斯^4,5,于飞^{1, 2},刘涛¹,刘朋朋¹(1.河北农业大学河北野生动物健康研究中心,河北保定 071000;2.河北农业大学生命科学学院,河北保定 071000;3.河北省科学院生物研究所,河北石家庄 050081;4.北京华科泰生物技术股份有限公司,北京 101111;5.天津华科泰生物技术有限公司,天津 300405)摘要:为检测乳腺癌患者血清人表皮生长因子受体2(HER2)蛋白,基于磁微粒化学发光免疫平台建立了血清HER2蛋白检测方法并评价其性能和市场产品等效性.结果显示:本研究建立的血清HER2蛋白检测方法检测范围为0~300 ng/mL;空白限(limit of blank, LoB)为0.48 ng/mL;外源性和内源性干扰平均回收率<5%;精密度变异系数(coefficient of variation,CV)<8%;稳定性实验显示,试剂可稳定储存48周,且与市售产品相关系数r为0.995;平均偏倚为-0.2 ng/mL.提示本研究成功建立了HER2蛋白检测方法,且各项分析性能表现优异,与在售化学发光试剂等效性良好,可为临床应用提供科学的数据参考.关键词:乳腺癌;HER2;化学发光法;性能评价;等效性中图分类号:R737.9 文献标志码:A 文章编号:1000-1565(2022)03-0281-07Establishment and performance evaluation of chemiluminescent immunoassay for serum HER2 proteinWANG Juan^1,2, CHENG Hua³, HU Chenguang^4,5, LIN Si^4,5, YU Fei^1,2, LIU Tao¹, LIU Pengpeng¹(1. Hebei Wildlife Health Center, Hebei Agricultural University, Baoding 071000, China;2. College of Life Science, Hebei Agricultural University, Baoding 071000, China;3. Institute of Biology, Hebei Academy of Sciences, Shijiazhuang 050081, China;4. Beijing Savant Biotechnology Co. Ltd., Beijing 101111, China;5. Tianjin Savant Biotechnology Co. Ltd., Tianjin 300405,China)Abstract: This study established magnetic particles chemiluminescence method for the detection of serum human epidermal growth factor receptor 2(HER2)protein in the patients with breast cancer, and evaluated its performance and equivalence with market products. The detection range of HER2 protein was from 0 to 300 ng/mL; limit of blank(LoB)was 0.48 ng/mL; average recovery rates of exogenous and endogenous interference was lower than 5%; precision was coefficient of variation(CV)lower than 8%; the stability test showed that the reagent could be stored stably for 48 weeks, and the correlation coefficient with commercial products was r=0.995, the mean bias was -0.2 ng/mL. These results indicated that the detection method of serum HER2 protein was successfully established, and had an excellent- 收稿日期:2022-01-12 基金项目:国家重点研发计划项目(2020YFC084310);河北省科学院科技攻关项目(21313) 第一作者:王娟(1989—),女,河北廊坊人,河北农业大学副教授,主要从事抗肿瘤药物筛选和相关机制研究.E-mail: wangjuan@hebau.edu.cn 通信作者:刘朋朋(1985—),男,河北赵县人,河北农业大学研究员,博士,主要从事体外诊断技术研究.E-mail: liupeng@cwbz.ac.cn第3期王娟等:血清HER2蛋白化学发光免疫检测方法的建立及评价performance compared with HER2 protein chemiluminescence reagents sold on the market.

Key words: Breast cancer, HER2, chemiluminescence, performance evaluation, equivalence

CLC Number:

R737.9

WANG Juan, CHENG Hua, HU Chenguang, LIN Si, YU Fei, LIU Tao, LIU Pengpeng. Establishment and performance evaluation of chemiluminescent immunoassay for serum HER2 protein[J]. Journal of Hebei University(Natural Science Edition), 2022, 42(3): 281-287.

References

[1] BRAY F, FERLAY J, SOERJOMATARAM I, et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2018, 68(6): 394-424.
[2] 黎立喜,马飞.乳腺癌筛查和早期诊断的血液生物学标志物[J].国际肿瘤学杂志, 2021, 48(2): 109-112. DOI:10.3760/cma.j.cn371439-20200629-00021.
[3] 中国抗癌协会乳腺癌专业委员会.中国抗癌协会乳腺癌诊治指南与规范(2021年版)[J].中国癌症杂志, 2021, 31(10): 954-1040. DOI:10.19401/j.cnki.1007-3639.2021.10.013.
[4] XUHONG J C, QI X W, ZHANG Y, et al. Mechanism, safety and efficacy of three tyrosine kinase inhibitors lapatinib, neratinib and pyrotinib in HER2-positive breast cancer[J]. Am J Cancer Res, 2019, 9(10): 2103-2119.
[5] OH D Y, BANG Y J. HER2-targeted therapies—a role beyond breast cancer[J]. Nat Rev Clin Oncol, 2020, 17(1): 33-48. DOI:10.1038/s41571-019-0268-3.
[6] 朱成斌,陈晓丽.免疫组化指标HER-2联合肿瘤标志物检测在胃癌诊断中的应用价值[J].慢性病学杂志, 2021, 22(10): 1573-1574, 1577. DOI:10.16440/J.CNKI.1674-8166.2021.10.36.
[7] BRANDT-RAUF P W, PINCUS M R, CARNEY W P. The c-erbB-2 protein in oncogenesis: molecular structure to molecular epidemiology[J]. Crit Rev Oncog, 1994, 5(2/3): 313-329. DOI:10.1615/critrevoncog.v5.i2-3.100.
[8] MORGAN S, AMEMIYA Y, SLODKOWSKA E, et al. Pilot study on the utility of circulating HER2/neu levels in the serum of breast cancer patients[J]. Anticancer Res, 2019, 39(10): 5345-5352. DOI:10.21873/anticanres.13727.
[9] 周爱清,陈志军,孙正魁,等.乳腺癌患者血清 HER-2/neu浓度与组织 HER-2表达水平的相关性[J].实用癌症杂志, 2015, 30(4): 479-481. DOI:10.3969/j.issn.1001-5930.2015.04.002.
[10] TSÉ C, GAUCHEZ A S, JACOT W, et al. HER2 shedding and serum HER2 extracellular domain: biology and clinical utility in breast cancer[J]. Cancer Treat Rev, 2012, 38(2): 133-142. DOI:10.1016/j.ctrv.2011.03.008.
[11] 王璐,沈国栋,顾朋颖,等.血清人表皮生长因子受体2水平在乳腺肿瘤诊断与治疗中的应用[J].中国临床保健杂志, 2020, 23(6): 820-822. DOI:10.3969/J.issn.1672-6790.2020.06.024.
[12] 刘永波,陈奕至,李艳萍,等. HER-2、CA153及HE4联合检测对乳腺癌的诊断价值[J].国际检验医学杂志, 2020, 41(2): 201-204. DOI:10.3969/j.issn.1673-4130.2020.02.018.
[13] 程灿灿,陈顺仪,梁紫甄,等.化学发光免疫分析法HER-2测定试剂盒的初步临床评价[J].标记免疫分析与临床, 2019, 26(8): 1385-1388, 1440. DOI:10.11748/bjmy.issn.1006-1703.2019.08.032.
[14] GOUTSOULIAK K, VEERARAGHAVAN J, SETHUNATH V, et al. Towards personalized treatment for early stage HER2-positive breast cancer[J]. Nat Rev Clin Oncol, 2020, 17(4): 233-250. DOI:10.1038/s41571-019-0299-9.
[15] MOELANS C B, DE WEGER R A, DER WALL E V, et al. Current technologies for HER2 testing in breast cancer[J]. Crit Rev Oncol, 2011, 80(3): 380-392. DOI:10.1016/j.critrevonc.2010.12.005.
[16] 张鹏宇,贾书生.新型血清标志物HER-2在乳腺癌诊疗中应用[J].中国实验诊断学, 2020, 24(8): 1367-1371. DOI:10.3969/j.issn.1007-4287.2020.08.044.
[17] TOI M, SPERINDE J, HUANG W D, et al. Differential survival following trastuzumab treatment based on quantitative HER2 expression and HER2 homodimers in a clinic-based cohort of patients with metastatic breast cancer[J]. BMC Cancer, 2010, 10: 56. DOI:10.1186/1471-2407-10-56.
[18] FABRICIO A S C, MICHILIN S, ZANCAN M, et al. Shed HER2 surrogacy evaluation in primary breast cancer patients: a study assessing tumor tissue HER2 expression at both extracellular and intracellular levels[J]. Scand J Clin Lab Investig, 2019, 79(4): 260-267. DOI:10.1080/00365513.2019.1600200.
[19] LEE S B, LEE J W, YU J H, et al. Preoperative serum HER2 extracellular domain levels in primary invasive breast cancer[J]. BMC Cancer, 2014, 14: 929. DOI:10.1186/1471-2407-14-929.
[20] 何永鹏,李丽仙,易琳,等.乳腺癌患者血清HER-2胞外域水平与癌组织HER-2表达的相关性及其临床意义[J/OL].中华乳腺病杂志, 2016, 10(1): 14-19. DOI:10.3877/cma.j.issn.1674-0807.2016.01.004. (