Optimizing antigen preparation for oxalyl-CoA decarboxylase enzyme diagnostic kit and ELISA system cutoff determination.
Cut off
ELISA
Oxalyl coA decarboxylase
Journal
Urolithiasis
ISSN: 2194-7236
Titre abrégé: Urolithiasis
Pays: Germany
ID NLM: 101602699
Informations de publication
Date de publication:
09 Oct 2024
09 Oct 2024
Historique:
received:
11
06
2024
accepted:
30
09
2024
medline:
9
10
2024
pubmed:
9
10
2024
entrez:
9
10
2024
Statut:
epublish
Résumé
The prevalence of kidney stone disease is increasing globally, with calcium oxalate stones being the most common type. Oxalyl-CoA decarboxylase (OXC), an enzyme produced by the gut bacterium Oxalobacter formigenes, plays a crucial role in oxalate metabolism. Deficiencies in OXC activity can lead to the accumulation of oxalate, contributing to kidney stone formation. This study aimed to develop a reliable diagnostic assay for OXC by optimizing antigen production and establishing a cutoff value for an enzyme-linked immunosorbent assay (ELISA). We cloned, expressed, and purified recombinant OXC protein in Escherichia coli BL21(DE3), and generated specific polyclonal antibodies in rabbits. The ELISA system was optimized and validated using serum samples from 40 healthy individuals and 6 patients with oxalate-related disorders. The cutoff value was determined using the formula (M + 2SD), where (M) is the mean and (SD) is the standard deviation of the healthy sample results. The calculated cutoff value of 0.656750 effectively distinguished between healthy and affected individuals, with a sensitivity of 97.5% and a specificity of 83.3%. These findings provide a valuable tool for the early detection and management of oxalate-related disorders, with significant implications for clinical practice.
Identifiants
pubmed: 39382754
doi: 10.1007/s00240-024-01635-7
pii: 10.1007/s00240-024-01635-7
doi:
Substances chimiques
oxalyl CoA decarboxylase
EC 4.1.1.8
Carboxy-Lyases
EC 4.1.1.-
Recombinant Proteins
0
Reagent Kits, Diagnostic
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
141Subventions
Organisme : The present study was carried out in the Cellular & Molecular Biology Research Center of Shahid Beheshti University of Medical Sciences, Tehran, Iran. and supported by Urology and Nephrology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
ID : 898
Informations de copyright
© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.
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