Fu Yulin, Wang Xiaoyan, Chen Xindong, Hong Jianfeng, Qin Yuan, Zhou Zixuan, Zhou Xiumei, Wang Yigang, Zhou Jingnan, Fang Hongming, Liu Pengfei, Huang Biao
College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, China.
Affiliated Xiaoshan Hospital, Hangzhou Normal University, Hangzhou, China.
Anal Biochem. 2023 Apr 1;666:115072. doi: 10.1016/j.ab.2023.115072. Epub 2023 Feb 3.
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.
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.
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).
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.
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。
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。
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）。
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。
