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基质金属蛋白酶3时间分辨免疫分析方法的建立及一些潜在的临床应用。

Establishment of matrix metalloproteinase 3 time-resolved immunoassay and some potential clinical applications.

作者信息

Fu Yulin, Wang Xiaoyan, Chen Xindong, Hong Jianfeng, Qin Yuan, Zhou Zixuan, Zhou Xiumei, Wang Yigang, Zhou Jingnan, Fang Hongming, Liu Pengfei, Huang Biao

机构信息

College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, China.

Affiliated Xiaoshan Hospital, Hangzhou Normal University, Hangzhou, China.

出版信息

Anal Biochem. 2023 Apr 1;666:115072. doi: 10.1016/j.ab.2023.115072. Epub 2023 Feb 3.

Abstract

AIM

To develop a highly sensitive time-resolved fluorescence immunoassay (TRFIA) for the detection of serum matrix metalloproteinase-3 (MMP-3) and to assess MMP-3's clinical value in patients with colorectal cancer (CRC).st.

METHODS

MMP-3 levels were established using the double antibody sandwich technique. The MMP-3 TRFIA technique was developed and optimized, and its linearity, sensitivity, accuracy, specificity, and recovery were assessed. Then, serum concentrations in healthy individuals and patients with CRC were determined by MMP-3 TRFIA.

RESULTS

The linear range of MMP-3 TRFIA was 0.73-500 ng/mL. MMP-3 TRFIA had an intra-batch precision range of 2.16%-7.10% percent and an inter-batch precision range of 3.99%-11.21%. MMP-3, tumor-associated trypsinogen 2, and AFP had no cross reaction.The recovery is between 90% and 110%, and had no serum interference. Patients with CRC had serum MMP-3 levels (73.95 ± 78.43 ng/mL) that were considerably higher than those of healthy individuals (21.45 ± 11.12 ng/mL), and those with metastasis had serum MMP-3 levels (95.89 ± 76.21 ng/mL) that were considerably higher than those of patients without metastasis (52.74 ± 47.25 ng/mL).

CONCLUSIONS

A highly sensitive MMP-3 TRFIA assay was successfully developed, and serum MMP-3 may be associated with CRC invasion and metastasis. Therefore, MMP-3 can be used in the auxiliary diagnosis of CRC.

摘要

目的

建立一种高灵敏度的时间分辨荧光免疫分析法(TRFIA)用于检测血清基质金属蛋白酶-3(MMP-3),并评估MMP-3在结直肠癌(CRC)患者中的临床价值。

方法

采用双抗体夹心技术测定MMP-3水平。开发并优化了MMP-3 TRFIA技术,并评估其线性、灵敏度、准确性、特异性和回收率。然后,通过MMP-3 TRFIA测定健康个体和CRC患者的血清浓度。

结果

MMP-3 TRFIA的线性范围为0.73 - 500 ng/mL。MMP-3 TRFIA的批内精密度范围为2.16% - 7.10%,批间精密度范围为3.99% - 11.21%。MMP-3、肿瘤相关胰蛋白酶原2和甲胎蛋白无交叉反应。回收率在90%至110%之间,且无血清干扰。CRC患者的血清MMP-3水平(73.95±78.43 ng/mL)显著高于健康个体(21.45±11.12 ng/mL),有转移的患者血清MMP-3水平(95.89±76.21 ng/mL)显著高于无转移的患者(52.74±47.25 ng/mL)。

结论

成功开发了一种高灵敏度的MMP-3 TRFIA检测方法,血清MMP-3可能与CRC的侵袭和转移有关。因此,MMP-3可用于CRC的辅助诊断。

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