A SIMPLER AND MORE SENSITIVE MODIFIED CATALYTIC (BLEOMYCIN DETECTABLE) IRON ASSAY.

IF 2.1 4区医学 Q3 MEDICAL LABORATORY TECHNOLOGY Annals of Clinical Biochemistry Pub Date : 2025-03-14 DOI:10.1177/00045632251330176

Mohan M Rajapurkar, Banibrata Mukhopadhyay, Suhas S Lele, Sudhir V Shah

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Abstract

Background: Iron is ubiquitously distributed in biology, only a miniscule amount exists as free is capable of catalysing production of highly toxic reactive hydroxyl radicle. This free iron also called; labile iron, non-transferrin bound iron or catalytic iron (CI). CI is measured by bleomycin-detectable iron assay. The assay as described originally was difficult to perform accurately and reproducibly due to variations of pH in the assay mixture and due to the lack of properly diluted iron standards.

Methods: . In our laboratory we modified the assay for serum/plasma so that the variations of pH in assay medium were constantly between 7.4 to 7.6 using acid diluted iron standards by multiple treatments of Chelex resin which is alkaline.

Results: Intra assay CV for low, medium, and high levels of catalytic iron was 0.05%, 0.61% and 0.68% whereas the interassay CV was 0.06%, 0.96% and .28% respectively. The modified assay is highly sensitive being able to detect levels as low as 0.1 μ mol/l. In patients on maintenance haemodialysis CI measured by the original assay failed to detect any catalytic iron in almost all of these samples whereas by modified method it was measurable in all patients with a mean of 0.66 ± 0.10 μ mol/l.. Normal values for catalytic iron in subjects having no comorbidities measured by modified method is 0.11± 0.06 µ moles/l.

Conclusions: The modified assay is reproducible and more sensitive than original assay and has been validated in several clinical studies.

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背景：铁在生物体内无处不在，只有极少量的游离铁能够催化产生毒性极强的活性羟基辐射。这种游离铁也称为易变铁、非转铁蛋白结合铁或催化铁（CI）。催化铁（CI）是通过博莱霉素可检测铁测定法测量的。由于化验混合物的 pH 值变化以及缺乏适当稀释的铁标准品，最初描述的化验很难准确地重复进行。我们实验室对血清/血浆检测方法进行了改进，使用酸性稀释的铁标准品，通过多次处理碱性的 Chelex 树脂，使检测介质的 pH 值始终保持在 7.4 至 7.6 之间：低、中、高水平催化铁的测定内 CV 分别为 0.05%、0.61% 和 0.68%，而测定间 CV 分别为 0.06%、0.96% 和 .28%。改进后的检测方法灵敏度高，可检测到低至 0.1 μ mol/l 的水平。在接受维持性血液透析的患者中，几乎所有样本都无法通过原始测定法检测到任何催化铁，而通过改良方法，所有患者都能检测到催化铁，平均值为 0.66 ± 0.10 μ mol/l。用改良方法测量无合并症患者的催化铁正常值为 0.11±0.06 μ moles/l：结论：改良测定法的重现性和灵敏度均优于原始测定法，并已在多项临床研究中得到验证。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Annals of Clinical Biochemistry Biochemistry, Genetics and Molecular Biology-Clinical Biochemistry

CiteScore

5.20

自引率

4.50%

发文量

期刊介绍： Annals of Clinical Biochemistry is the fully peer reviewed international journal of the Association for Clinical Biochemistry and Laboratory Medicine. Annals of Clinical Biochemistry accepts papers that contribute to knowledge in all fields of laboratory medicine, especially those pertaining to the understanding, diagnosis and treatment of human disease. It publishes papers on clinical biochemistry, clinical audit, metabolic medicine, immunology, genetics, biotechnology, haematology, microbiology, computing and management where they have both biochemical and clinical relevance. Papers describing evaluation or implementation of commercial reagent kits or the performance of new analysers require substantial original information. Unless of exceptional interest and novelty, studies dealing with the redox status in various diseases are not generally considered within the journal''s scope. Studies documenting the association of single nucleotide polymorphisms (SNPs) with particular phenotypes will not normally be considered, given the greater strength of genome wide association studies (GWAS). Research undertaken in non-human animals will not be considered for publication in the Annals. Annals of Clinical Biochemistry is also the official journal of NVKC (de Nederlandse Vereniging voor Klinische Chemie) and JSCC (Japan Society of Clinical Chemistry).