双粒径胶乳免疫比浊法在测定D- 二聚体中的应用

doi:10.3969/j.issn.1673-8640.2016.09.015

摘要/Abstract

摘要：

目的改进胶乳免疫试剂的分析灵敏度和线性范围,并验证在用胶乳免疫比浊法测定D-二聚体（DD）的方法中,采用双粒径胶乳致敏DD单克隆抗体比用单一粒径胶乳具有更好的灵敏度和线性范围。方法制备2种平均粒径不同的羧基化胶乳微粒（49和150 nm）,并采用化学交联法偶联抗DD单克隆抗体,将2种粒径的致敏胶乳按不同比例混合,测定其分析灵敏度和线性范围,并与市售进口试剂进行比对。结果当49和150 nm致敏胶乳按4:1的比例混合时,其测定的校准曲线线性最好,线性范围达到61.88 μg/mL,分析灵敏度达到0.03,与进口试剂检测结果具有良好的相关性。结论双粒径致敏胶乳制备的DD免疫比浊法试剂比市售的传统DD胶乳诊断试剂的性能有了一定的提高,该试剂具有良好的临床应用前景。

关键词: D-二聚体, 免疫比浊法, 双粒径胶乳, 灵敏度, 线性范围, 全自动血凝仪

Abstract:

Objective To improve the analytical sensitivity and linear range of latex immunological reagent,and to compare the sensitivities and linear ranges of dual and single particle size latex immunoturbidimetry in the determination of D-dimer（DD）. Methods Carboxylic latex particles with diameters of 49 and 150 nm were prepared,and a covalent coupling method was used to link DD monoclonal antibody to the particles. The 2 sensitized latex particles were mixed at various proportions,and the sensitivity and linear range were determined,which were compared with those of imported reagents. Results If 4 parts of 49 nm sensitized latex particles were mixed with 1 part of 150 nm sensitized latex particles,the optimal linearity of calibration curve was obtained,with the sensitivity of 0.03 and linear range of 61.88 μg/mL,and it correlated well with imported reagents. Conclusions Dual particle size latex immunoturbidimetry is better than traditional DD latex diagnostic assay,and is of potential for clinical application.

Key words: D-dimer, Immunoturbidimetry, Dual particle size latex, Sensitivity, Linear range, Automatic blood coagulation analyzer

中图分类号:

R446.1

方清. 双粒径胶乳免疫比浊法在测定D- 二聚体中的应用[J]. 检验医学, 2016, 31(9): 801-806.

FANG Qing. The determination of D-dimer by dual particle size latex immunoturbidimetry[J]. Laboratory Medicine, 2016, 31(9): 801-806.

图/表 10

参考文献 7

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胶乳浓度（mg/mL）	回归方程	R²值
3	Y = 0.000 6X + 0.000 4	0.997 1
4	Y = 0.001 3X + 0.000 7	0.997 8
5	Y = 0.001 9X + 0.000 7	0.998 3
6	Y = 0.002 5X + 0.000 8	0.998 9
7	Y = 0.002 6X + 0.000 7	0.999 3
8	Y = 0.002 9X + 0.000 6	0.999 6
9	Y = 0.003 2X + 0.003 5	0.976 2

胶乳浓度（mg/mL）	回归方程	R²值
3	Y = 0.000 6X + 0.000 4	0.997 1
4	Y = 0.001 3X + 0.000 7	0.997 8
5	Y = 0.001 9X + 0.000 7	0.998 3
6	Y = 0.002 5X + 0.000 8	0.998 9
7	Y = 0.002 6X + 0.000 7	0.999 3
8	Y = 0.002 9X + 0.000 6	0.999 6
9	Y = 0.003 2X + 0.003 5	0.976 2

胶乳浓度（mg/mL）	回归方程	R²值
1	Y = 0.006 6X -0.001 7	0.999 4
2	Y = 0.007 8X -0.002 8	0.997 7
3	Y = 0.008 6X -0.003 1	0.996 9
4	Y = 0.007 4X + 0.004 3	0.987 9

胶乳浓度（mg/mL）	回归方程	R²值
1	Y = 0.006 6X -0.001 7	0.999 4
2	Y = 0.007 8X -0.002 8	0.997 7
3	Y = 0.008 6X -0.003 1	0.996 9
4	Y = 0.007 4X + 0.004 3	0.987 9

49 nm:150 nm	回归方程	R²值
1:1	Y = 0.008 1X + 0.006 1	0.989 3
1:2	Y = 0.008 6X + 0.016 4	0.931 2
1:3	Y = 0.007 6X + 0.027 6	0.810 4
1:4	Y = 0.008 2X + 0.028 9	0.797 6
2:1	Y = 0.005 4X -0.003 4	0.992 0
3:1	Y = 0.006 6X -0.001 2	0.999 6
4:1	Y = 0.007 2X + 0.001 6	0.999 7