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Updated: Aug 2, 2026
A Modified Precipitation Method to Isolate Urinary Exosomes
Published on: January 16, 2015
Measurement of urinary exosomal phospholipase A2 receptor is a sensitive method for diagnosis of PLA2R-associated
Bin Wang1, Yu-Qi Fu1, Li-Jun Xie1
1Institute of Nephrology, Zhong Da Hospital, Southeast University School of Medicine, Nanjing, Jiangsu, China.
Background:
The discovery of phospholipase A2 receptor (PLA2R) and its antibody (aPLA2Rab) has paved the way for diagnosing PLA2R-associated membranous nephropathy (PLA2R-MN) with a high specificity of 98%. However, the sensitivity was only 40% to 83.9%, and there is ongoing discussion around determining the optimal threshold for diagnosis. Recent advancements in the use of exosomes, a novel form of "liquid biopsy," have shown great promise in identifying markers for various medical conditions.
Methods:
Protein mass spectrometry and western blot were applied to verify the existence of PLA2R antigen in the urine exosome. We then evaluated the efficacy of urinary exosomal PLA2R antigen alone or combined with serum aPLA2Rab level to diagnose PLA2R-MN.
Results:
The urinary exosomes contained a high abundance of PLA2R antigen as evidenced by protein mass spectrometry and western blot in 85 PLA2R-MN patients vs the disease controls (14 secondary MN patients, 22 non-MN patients and 4 PLA2R-negative MN patients) and 20 healthy controls. Of note, urinary exosomal PLA2R antigen abundance also had a good consistency with the PLA2R antigen level in the renal specimens of PLA2R-MN patients. The sensitivity of urinary exosomal PLA2R for diagnosing PLA2R-MN reached 95.4%, whereas the specificity was 63.3%. Combining detection of the urinary exosomal PLA2R and serum aPLA2Rab could develop a more sensitive diagnostic method for PLA2R-MN, especially for patients with serum aPLA2Rab ranging from 2 to 20 RU/mL.
Conclusions:
Measurement of urinary exosomal PLA2R could be a sensitive method for the diagnosis of PLA2R-MN.
Insights
Urinary exosomal phospholipase A2 receptor (PLA2R) antigen shows high sensitivity for diagnosing PLA2R-associated membranous nephropathy (PLA2R-MN). Combining this with serum antibody detection offers a more sensitive diagnostic approach for PLA2R-MN.
Area of Science:
- Nephrology
- Biomarker Discovery
- Exosome Biology
Background:
- Phospholipase A2 receptor (PLA2R) and its antibody (aPLA2Rab) are key diagnostic markers for PLA2R-associated membranous nephropathy (PLA2R-MN).
- Current diagnostic methods for PLA2R-MN exhibit high specificity but variable sensitivity (40-83.9%).
- Exosomes, as a form of liquid biopsy, offer potential for identifying novel diagnostic markers.
Purpose of the Study:
- To investigate the presence and diagnostic utility of urinary exosomal PLA2R antigen in PLA2R-MN.
- To evaluate the combined diagnostic efficacy of urinary exosomal PLA2R antigen and serum aPLA2Rab.
Main Methods:
- Protein mass spectrometry and western blot were used to detect PLA2R antigen in urinary exosomes.
- The diagnostic performance of urinary exosomal PLA2R antigen, alone and in combination with serum aPLA2Rab, was assessed in PLA2R-MN patients and controls.
Main Results:
- Urinary exosomes from PLA2R-MN patients showed a high abundance of PLA2R antigen.
- Urinary exosomal PLA2R antigen demonstrated high sensitivity (95.4%) but moderate specificity (63.3%) for diagnosing PLA2R-MN.
- The abundance of urinary exosomal PLA2R antigen correlated with renal specimen levels.
- Combining urinary exosomal PLA2R and serum aPLA2Rab improved diagnostic sensitivity, particularly for patients with serum aPLA2Rab levels between 2-20 RU/mL.
Conclusions:
- Urinary exosomal PLA2R antigen is a highly sensitive biomarker for PLA2R-MN diagnosis.
- This exosomal marker, especially when combined with serum aPLA2Rab, enhances diagnostic accuracy for PLA2R-MN.
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