ELISA analysis of urinary nephrin and podocalyxin standardized by aquaporin-2 in adult patients with nephrotic syndrome

J Nephrol. 2014 Aug;27(4):411-7. doi: 10.1007/s40620-014-0066-z. Epub 2014 Apr 16.

Abstract

Objective: To investigate urinary nephrin and podocalyxin standardized by aquaporin (AQP)-2 using the enzyme-linked immunosorbent assay (ELISA) method in adult nephrotic syndrome (NS) patients.

Methods: In 107 adult NS patients (27 proliferative nephritis, 77 non-proliferative, and 3 amyloidosis) undergoing renal biopsy, urinary nephrin, podocalyxin and AQP2 were measured by ELISA. Urinary nephrin and podocalyxin were standardized by AQP2 (neph/AQP and PCX/AQP) and values were compared with 11 healthy controls.

Results: Urinary neph/AQP correlated positively to PCX/AQP (r = 0.51, p < 0.001). Urinary neph/AQP and PCX/AQP were lower in controls than NS patients. Both proliferative and non-proliferative NS patients excreted high urinary neph/AQP and PCX/AQP without a significant difference between them (p > 0.05). Patients with focal segmental glomerular sclerosis (FSGS) excreted higher urinary neph/AQP (p = 0.09) and PCX/AQP (p < 0.05) compared to the other patients. Urinary neph/AQP and PCX/AQP were increased in the immunoglobulin M nephropathy patients. Amyloidosis patients excreted lower neph/AQP and PCX/AQP. The sensitivity was 0.87 and specificity 0.37 when the neph/AQP borderline value of 0.16 was adopted [area under the curve (AUC) = 0.61]. The sensitivity was 0.74 and specificity 0.61 when the PCX/AQP borderline value was 3.06 (AUC = 0.69).

Conclusions: Urinary neph/AQP and PCX/AQP are increased in NS patients, with FSGS patients showing the highest levels. To distinguish FSGS from other NS forms, the measurement of urinary PCX/AQP may be a practical method, and superior to neph/AQP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Amyloidosis / urine*
  • Aquaporin 2 / urine*
  • Case-Control Studies
  • Child
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Glomerulonephritis / urine*
  • Humans
  • Male
  • Membrane Proteins / urine*
  • Middle Aged
  • Nephrosis, Lipoid / urine
  • Nephrotic Syndrome / pathology
  • Nephrotic Syndrome / urine*
  • Podocytes*
  • Sensitivity and Specificity
  • Sialoglycoproteins / urine*
  • Urine / cytology
  • Young Adult

Substances

  • Aquaporin 2
  • Membrane Proteins
  • Sialoglycoproteins
  • nephrin
  • podocalyxin