双光POCT型乳胶免疫比浊法测定糖化血红蛋白关键技术的研究

doi:10.19586/j.2095-2341.2024.0162

生物技术进展 ›› 2025, Vol. 15 ›› Issue (2): 333-340.DOI: 10.19586/j.2095-2341.2024.0162

• 研究论文 • 上一篇

双光POCT型乳胶免疫比浊法测定糖化血红蛋白关键技术的研究

贾艳艳¹(), 郭红壮²()

^1.吉林医药学院附属医院，吉林吉林 132000
^2.长春理工大学物理学院，长春 130000

收稿日期:2024-10-22 接受日期:2024-11-05 出版日期:2025-03-25 发布日期:2025-04-29
通讯作者: 郭红壮
作者简介:贾艳艳 E-mail: 1019570283@qq.com；

Study on the Key Technology for the Determination of Glycated Haemoglobin by Double-light POCT-type Latex Immunoturbidimetric Assay

Yanyan JIA¹(), Hongzhuang GUO²()

^1.Affiliated Hospital of Jilin Medical College，Jilin Jilin 132000，China
^2.School of Physics，Changchun University of Science and Technology，Changchun 130000，China

Received:2024-10-22 Accepted:2024-11-05 Online:2025-03-25 Published:2025-04-29
Contact: Hongzhuang GUO

摘要/Abstract

摘要：

糖尿病已成为危害人体健康的一种常见疾病，糖化血红蛋白由于不受采样时血糖浓度影响，因而检测糖化血红蛋白浓度对病情的监控更具有意义。依据朗伯比尔定律，利用光对物质选择性吸收的特性，设计了一款双光POCT型乳胶免疫比浊法测定糖化血红蛋白的装置，对其激发光路、孵育工位等主要部件进行了建模仿真，对仪器的主要性能进行了验证，并与UV-1780分光光度计进行了仪器对比实验。结果表明，装置的线性相关性R²=0.998 4，重复性≤ 0.997 9%，孵育工位的平均升温速度为0.44 ℃·s^-1，温度均匀性≤ ± 0.1 ℃（温度为37.3 ℃）。与UV-1780分光光度计测量21种不同浓度的糖化血红蛋白抗凝血阳性质控品样本的趋势一致，充分证明了本装置的检测结果准确、可靠，可为糖化血红蛋白检测装置的POCT化提供了前期的实验基础。

关键词: 乳胶免疫比浊法, 糖化血红蛋白, 有限元分析, POCT, 光电比浊法

Abstract:

Diabetes mellitus has become a common disease that endangers human health. Glycated haemoglobin is not affected by the concentration of blood glucose at the time of sampling， so its concentration detection is more meaningful for monitoring the disease. In this paper， based on Lambert's Beer's law， using the characteristic of selective absorption of light on substances， we designed a two-optical POCT-type latex immunoturbidimetric device for the determination of glycated haemoglobin， and carried out modelling simulation of its excitation optical path， incubation station and other main components， performance verification of the main performance of the instrument， and instrument comparison experiments with UV-1780 spectrophotometer. The experimental results showed that the linear correlation R²=0.998 4， the repeatability≤0.997 9%， the average heating rate of the incubation station was 0.44 ℃·s^-1， and the temperature uniformity was ≤±0.1 ℃ （the temperature was 37.3 ℃）. The trend was consistent with that of the UV-1780 spectrophotometer， which measured 21 different concentrations of glycated haemoglobin anticoagulant-positive QC samples， fully proving that the detection results of this device were accurate and reliable， and providing a pre-experimental basis for the POCT of the glycated haemoglobin detection device.

Key words: latex immunoturbidimetric assay, glycated haemoglobin, finite element analysis, POCT, phototurbidimetric method

中图分类号:

R318.6

贾艳艳, 郭红壮. 双光POCT型乳胶免疫比浊法测定糖化血红蛋白关键技术的研究[J]. 生物技术进展, 2025, 15(2): 333-340.

Yanyan JIA, Hongzhuang GUO. Study on the Key Technology for the Determination of Glycated Haemoglobin by Double-light POCT-type Latex Immunoturbidimetric Assay[J]. Current Biotechnology, 2025, 15(2): 333-340.

图/表 15

参考文献 16

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试剂盒组成		试剂中主要成分	含量
试剂1	R1	乳胶	60 mL·L^-1
试剂1	R1	甘氨酸缓冲液	0.1 mL·L^-1
试剂2	R2a	羊抗鼠IgG抗体	50 mg·L^-1
	R2a	甘氨酸缓冲液	0.1 mL·L^-1
	R2b	鼠抗人HbA1c单克隆抗体	120 mg·L^-1
	R2b	甘氨酸缓冲液	0.1 mL·L^-1
溶血液		H₂O	100 mL

试剂盒组成		试剂中主要成分	含量
试剂1	R1	乳胶	60 mL·L^-1
试剂1	R1	甘氨酸缓冲液	0.1 mL·L^-1
试剂2	R2a	羊抗鼠IgG抗体	50 mg·L^-1
	R2a	甘氨酸缓冲液	0.1 mL·L^-1
	R2b	鼠抗人HbA1c单克隆抗体	120 mg·L^-1
	R2b	甘氨酸缓冲液	0.1 mL·L^-1
溶血液		H₂O	100 mL

序号	实验步骤	备注
1	向比色皿中加入试剂1	300 μL
2	向比色皿中加入样本	8 μL
3	混匀后放入装置，孵育5 min
4	向比色皿中加入试剂2	100 μL
5	混匀后,孵育1 min后读取吸光度A1，再孵育5 min后读取吸光度A2
6	记录ΔA=A2-A1

序号	实验步骤	备注
1	向比色皿中加入试剂1	300 μL
2	向比色皿中加入样本	8 μL
3	混匀后放入装置，孵育5 min
4	向比色皿中加入试剂2	100 μL
5	混匀后,孵育1 min后读取吸光度A1，再孵育5 min后读取吸光度A2
6	记录ΔA=A2-A1

浓度(%，NGSP)	UV-1780吸光度	本装置吸光度
0.00	0.000 0	0.022 7
4.51	0.022 9	0.060 9
7.11	0.050 9	0.116 6
9.94	0.081 9	0.164 8
14.75	0.132 7	0.241 5

双光POCT型乳胶免疫比浊法测定糖化血红蛋白关键技术的研究

Study on the Key Technology for the Determination of Glycated Haemoglobin by Double-light POCT-type Latex Immunoturbidimetric Assay

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图/表 15

参考文献 16

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序号	UV-1780	本装置
1	0.037 1	0.060 2
2	0.037 3	0.060 7
3	0.033 7	0.060 1
4	0.036 9	0.059 1
5	0.037 6	0.059 7
平均值	0.037 26	0.059 96
标准差	0.000 27	0.000 60
CV	0.725 1%	0.997 9%

序号	UV-1780	本装置	序号	UV-1780	本装置
1	-0.009 2	-0.006 1	12	-0.001 6	0.009 2
2	0.082 8	0.264 1	13	0.072 7	0.175 4
3	0.078 6	0.191 2	14	0.069 7	0.167 8
4	-0.009 9	-0.007 5	15	0.042 3	0.103 2
5	0.008 1	0.029 3	16	0.011 3	0.036 0
6	0.057 8	0.138 8	17	0.056 5	0.135 6
7	-0.008 7	-0.005 1	18	0.084 9	0.209 1
8	0.087 7	0.265 3	19	0.059 5	0.142 7
9	0.092 7	0.263 2	20	0.066 5	0.253 0
10	0.026 5	0.068 4	21	-0.009 7	-0.007 2
11	-0.003 6	0.005 1

	本装置	UV-1780	Uicare-300
样品通量	1	6	32
灵敏度	0.003 Abs	0.001 Abs	0.01 Abs
检测范围	0.01~2.0 mg·L^-1	0.01~2.0 mg·L^-1	0.1~2.0 mg·L^-1
优势	价格低、体积小、操作简便	激发波长在340~900 nm内，以1 nm步进	样本通量相对较大，可以自动加样