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. 2022 Sep;36(9):e24603.
doi: 10.1002/jcla.24603. Epub 2022 Jul 23.

Establishment of a time-resolved immunoassay for acute kidney injury based on the detection of Kim-1

Zheng Shaoxiong  1 , Xiumei Zhou  1 , Yuan Qin  1 , Yu Xiaomei  1 , Chen Lingli  1 , Liu Xiaobin  2 , Yigang Wang  1 , Gong Jianguang  3 , Shen Shuijuan  4 , Biao Huang  1
Affiliations

Establishment of a time-resolved immunoassay for acute kidney injury based on the detection of Kim-1

Zheng Shaoxiong et al. J Clin Lab Anal. 2022 Sep.

Abstract

Aim: To establish a highly sensitive time-resolved fluorescence immunoassay (TRFIA) of kidney injury molecule-1 (Kim-1) and evaluate its clinical value in acute kidney injury (AKI).

Methods: The Kim-1-TRFIA was established by the double-antibody sandwich method, and the method was evaluated. The established Kim-1-TRFIA was used to detect the concentration of Kim-1 in the serum of healthy controls and patients with AKI.

Results: The optimal coating antibody concentration and optimal Eu3+ -labeled antibody dilution ratio for Kim-1-TRFIA are 1 μg/ml and 1:140, respectively. The linear range is 42.71-4666.69 pg/ml. The intra- and inter-assay coefficients of variation are <10%. The specificity of our Kim-1-TRFIA is acceptable. The recovery is between 95.14% and 102.84%. The concentration of Kim-1 in the serum of patients with AKI is 126.50 ± 67.99 pg/ml, which is significantly higher than that in the serum of healthy controls (49.72 ± 16.40 pg/ml, p < 0.001). Staging patients with AKI by glomerular filtration rate shows that the serum concentration of Kim-1 increases significantly with increasing disease severity (p < 0.05).

Conclusion: A highly sensitive Kim-1-TRFIA was established. With this immunoassay, a good differential diagnosis can be made, and healthy people and AKI patients can be differentiated by detecting the concentration of Kim-1 in the serum. Moreover, the severity of AKI patients can be determined.

Keywords: acute kidney injury; double-antibody sandwich method; kidney injury molecule-1; time-resolved fluorescence immunoassay.

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Figures

FIGURE 1
FIGURE 1
Experimental principle of the Kim‐1‐TRFIA method
FIGURE 2
FIGURE 2
Optimization of Kim‐1 coating concentration
FIGURE 3
FIGURE 3
Optimization of the dilution ratios of Eu3+–Kim‐1‐McAb
FIGURE 4
FIGURE 4
Optimization of Kim‐1‐TRFIA reaction time
FIGURE 5
FIGURE 5
Standard curve of the Kim‐1 standard
FIGURE 6
FIGURE 6
Correlation of Kim‐1 concentration results between ELISA and the newly established Kim‐1‐TRFIA
FIGURE 7
FIGURE 7
Kim‐1 concentration in patients with AKI patients and healthy controls
FIGURE 8
FIGURE 8
ROC curves of normal controls and AKI patients
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