The first paragraph is an introduction. The quantitative determination of urinary Cystatin C (cyst-C) in conjunction with the qualitative study of its polymorphisms is an excellent tool for detecting newborns who are predisposed to renal function deficiency early on. The immunometric methods PETIA, PENIA, and EIA are used to quantify cyst-C in human biologic fluid, but they have limitations and do not allow for qualitative study. The aim of this study is to prove that Immunoblot SDS-PAGE can be used to analyse urinary cyst-C in both qualitative and quantitative ways. Urine was obtained from newborns in the S. Maria della Misericordia Hospital nursey. The immunoblot SDS-PAGE and optical density reading were used to investigate urinary cyst-C.
The qualitative analysis revealed two molecular forms: a reactivity around 70 KDa in all samples and a reactivity around 13 KDa in a small number of samples. This research helps researchers to link cyst-C polymorphisms to specific changes in renal function in newborns. The quantitative analysis is targeted, sensitive, and precise.In reality, the assay precision and accuracy coefficients of variance were both 10%, the detection limit was 0.009 ng/L, and the calibration line had adequate linearity (range 0.02-0.3 ng/L). Also without the use of protease inhibitors, the stability of urinary cyst-C was sufficient as long as the assay was performed on newly recruited urine or immediately after thawing samples that had been frozen for up to six months.
Conclusion: Immunoblot SDS-PAGE study of urinary cyst-C is a reliable tool for obtaining a qualitative and quantitative analysis. This method reveals previously unknown knowledge about a 70 KDa cyst-C shape. The assay could provide diagnostic information that isn't accessible through immunometric methods.
Author (s) Details
C. Grasselli
Department of Surgical and Biomedical Sciences, Section of Obstetrics and Gynecology, University of Perugia, Perugia, Italy.
A. Barbati
Department of Surgical and Biomedical Sciences, Section of Obstetrics and Gynecology, University of Perugia, Perugia, Italy.
L. Cesarini
Department of Chemistry, Biology and Biotechnology, University of Perugia, Perugia, Italy.
R. Pellegrino
Department of Chemistry, Biology and Biotechnology, University of Perugia, Perugia, Italy.
G. C. Di Renzo
Department of Surgical and Biomedical Sciences, Section of Obstetrics and Gynecology, University of Perugia, Perugia, Italy and GEBISA, Research Foundation, Perugia, Italy.
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