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通过 ELISA 方法设计草酰辅酶 A 脱羧酶诊断试剂盒。

Designing a diagnostic kit for Oxalyl CoA Decarboxylase enzyme by ELISA method.

机构信息

Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Urology and Nephrology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

出版信息

Immunol Lett. 2019 Jan;205:78-83. doi: 10.1016/j.imlet.2018.11.008. Epub 2018 Nov 15.

Abstract

Urinary stones are the third most commonly reported urinary tract disease. Kidney stones are one of the most common types of urinary stones that most of them (70-80%) are calcium oxalate. Oxalic acid is highly oxidized organic compounds, which found in dietary and produced by the intestinal microflora. Oxalyl CoA decarboxylase is a key enzyme and plays an important role in oxalate degradation. In this study, the Oxalyl CoA decarboxylase gene which contains an -histidine tag was cloned in pET 28a (+) and expressed in Escherichia coli BL21 (DE3). The purified Oxalyl CoA decarboxylase protein was injected into rabbit for immunization. The antibody against Oxalyl CoA decarboxylase protein was used in ELISA assy. eventually, this ELISA system was used for patients with calcium oxalate kidney stones. ELISA analysis of serum samples of patient white calcium oxalate kidney stones showed that 88.8% of patient was lacking in antibody against Oxalyl CoA decarboxylase. This study suggests that antibodies against oxalyl-co-decoxylase proteins are useful in the detection of urinary tract stones and It can be used to measure oxalyl CoAdecoxylase enzymes in a simple method.

摘要

尿路结石是第三大常见的尿路疾病。肾结石是最常见的尿路结石类型之一,其中大多数(70-80%)是草酸钙结石。草酸是高度氧化的有机化合物,存在于饮食中,并由肠道微生物群产生。草酰辅酶 A 脱羧酶是一种关键酶,在草酸降解中发挥重要作用。在这项研究中,含有 -组氨酸标签的草酰辅酶 A 脱羧酶基因被克隆到 pET 28a(+)中,并在大肠杆菌 BL21(DE3)中表达。纯化的草酰辅酶 A 脱羧酶蛋白被注射到兔子中进行免疫。针对草酰辅酶 A 脱羧酶蛋白的抗体用于 ELISA 测定。最终,该 ELISA 系统用于检测草酸钙肾结石患者。对患有白色草酸钙肾结石的患者血清样本进行 ELISA 分析表明,88.8%的患者缺乏针对草酰辅酶 A 脱羧酶的抗体。这项研究表明,针对草酰辅酶 A 脱羧酶蛋白的抗体可用于检测尿路结石,并可用于以简单的方法测量草酰辅酶 A 脱羧酶酶。

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