Establishment of a time‐resolved immunoassay for acute kidney injury based on the detection of Kim‐1 |
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Authors: | Zheng Shaoxiong Xiumei Zhou Yuan Qin Yu Xiaomei Chen Lingli Liu Xiaobin Yigang Wang Gong Jianguang Shen Shuijuan Biao Huang |
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Affiliation: | 1. College of Life Sciences and Medicine, Zhejiang Sci‐Tech University, Hangzhou China ; 2. Wuxi People''s Hospital affiliated to Nanjing Medical University, Wuxi China ; 3. Zhejiang Provincial People''s Hospital, Hangzhou China ; 4. Nephrology Department of Shaoxing People''s Hospital, Shaoxing Hospital, Zhejiang University School of Medicine, Hangzhou China |
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Abstract: | AimTo 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).MethodsThe 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.ResultsThe 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).ConclusionA 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. |
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Keywords: | acute kidney injury, double‐ antibody sandwich method, kidney injury molecule‐ 1, time‐ resolved fluorescence immunoassay |
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